AIM:To investigate the effect ofβ-alanine(BA)on laserinduced choroidal neovascularization(CNV)mice models.METHODS:Laser-induced CNV mice models were established,and BA was administrated for one week and two weeks in ...AIM:To investigate the effect ofβ-alanine(BA)on laserinduced choroidal neovascularization(CNV)mice models.METHODS:Laser-induced CNV mice models were established,and BA was administrated for one week and two weeks in advance,separately.Furthermore,retinal pigment epithelium(RPE)-choroid flat mounts were separated,and immunohistochemical staining was performed.The laser-induced CNV lesion areas were measured and compared.In addition,liver and kidney morphologies were observed to identify potential hepatorenal toxicity.RESULTS:Enlarged CNV lesion areas were observed in the BA treated group.No significant differences were observed in the liver and kidney sections between groups.CONCLUSION:BA treatment increase CNV lesion areas,suggesting the detrimental effects of BA as a nutritional supplement in age-related macular degeneration(AMD)population.展开更多
AIM:To observe the long-term clinical efficacy of intravitreal injections of conbercept,a novel vascular growth factor inhibitor,for the treatment of pathological myopia choroidal neovascularization(PM-CNV).METHODS:A ...AIM:To observe the long-term clinical efficacy of intravitreal injections of conbercept,a novel vascular growth factor inhibitor,for the treatment of pathological myopia choroidal neovascularization(PM-CNV).METHODS:A total of 67 eyes(from 67 patients;mean age,54.90±12.7y)with PM-CNV were retrospectively researched.Based on the different schemes used for the administration of the drug,the patients were divided into two groups:group A(n=35;average age,53.31±13.6y;average diopter,9.25±1.72 D),which received only one injection of pro re nata(PRN;1+PRN regimen),and group B(n=32;average age,56.49±11.8y;average diopter,9.63±2.24 D),which received one injection per month for 3mo(3+PRN regimen).Best-corrected visual acuity(BCVA)analysis,intraocular pressure(IOP)examination,slit-lamp microscopy,fundus examination and optical coherence tomography were per formed at each follow-up.The recurrence and treatment times of CNV were recorded.The patients were followed up for at least 12mo.RESULTS:The BCVA was increased in 29 eyes(82.9%)in group A and 30 eyes(93.75%)in group B;no increase or decrease was observed in 6(17.1%)and 2(6.25%)eyes in groups A and B,respectively.The BCVA(log MAR)values before treatment(0.67±0.48 and 0.71±0.56)were significantly higher than those 12mo after treatment(0.31±0.26 and 0.33±0.17)in groups A and B,respectively(P<0.05).The mean central macular thickness(CMT)values had significantly decreased from 346.49±65.99 and 360.10±82.31μm at baseline to 257.29±40.47 and 251.97±48.26μm in groups A and B,respectively,after 12mo of treatment.A total of 21 eyes in group A needed reinjection(60%;average number of injections,2.51±0.98);the corresponding values in group B were 6 eyes(18.75%;average number of injections,3.74±1.22).There were no adverse ocular and systemic complications during the treatment and follow-up.CONCLUSION:Intravitreal injection of conbercept with 1+PRN or 3+PRN improve the visual acuity,reduce macular edema and reduce the level of CMT in patients with PM-CNV.The 3+PRN regimen demonstrates a lower recurrence rate of CNV than the 1+PRN regimen,but requires more treatment.However,both treatment regimens demonstrate long-term safety and efficacy for the treatment of PM-CNV.展开更多
AIM:To investigate whether anti-placental growth factor(PGF) can inhibit subretinal fibrosis and whether this effect is mediated by the inhibitory effect of PGF on epithelial-mesenchymal transition(EMT) of retinal pig...AIM:To investigate whether anti-placental growth factor(PGF) can inhibit subretinal fibrosis and whether this effect is mediated by the inhibitory effect of PGF on epithelial-mesenchymal transition(EMT) of retinal pigment epithelial(RPE) cells.METHODS:Subretinal fibrosis model was established in laser induced choroidal neovascularization(CNV) mice on day 21 after laser photocoagulation.Immunofluorescence staining(IFS) of cryosections and enzyme-linked immunosorbent assay(ELISA) were used to detect the expression of PGF.IFS of whole choroidal flat-mounts was used to detect the degree of subretinal fibrosis.IFS of cryosections and ELISA were used to detect the expression of EMT related indicators in subretinal fibrosis lesions.RESULTS:The expression of PGF protein in subretinal fibrosis lesions was significantly up-regulated(P<0.05),and mainly co-stained with pan-cytokeratin labeled RPE cells.Intravitreal injection of anti-PGF neutralizing antibody reduced the area of subretinal fibrosis and the ratio of fibrotic/angiogenic area significantly at the concentrations of 0.25,0.5,1.0,and 2.0 μg/μL(all P<0.05).The expression of E-cadherin in the local RPE cells decreased,while α-SMA increased significantly in subretinal fibrosis lesions,and the application of anti-PGF neutralizing antibody could reverse these changes(P<0.05).CONCLUSION:The expression of PGF is up-regulated in the lesion site of subretinal fibrosis and mainly expressed in RPE cells.Intravitreal injection of anti-PGF neutralizing antibody can significantly inhibit the degree of subretinal fibrosis in CNV mice,and this effect may be mediated by the inhibition of PGF on EMT of RPE cells.展开更多
The present study reports a case of a patient with choroidal neovascularization(CNV)associated with pseudoxanthoma elasticum(PXE).We observed the functional and anatomical improvement of the patient treated with intra...The present study reports a case of a patient with choroidal neovascularization(CNV)associated with pseudoxanthoma elasticum(PXE).We observed the functional and anatomical improvement of the patient treated with intravitreal vascular endothelial growth factor(VEGF)inhibitor bevacizumab.The study also systematically searched the database for similar cases to provide a literature review.Data concerning the clinical features,treatment strategies and outcomes were extracted and analyzed.Retrospective interventional case report and systematic literature review.A 56-year-old healthy Chinese woman with CNV secondary to PXE was reported.Examinations included best corrected visual acuity(BCVA),biomicroscopy,optical coherence tomography(OCT),fluorescein and indocyanine green angiography and digital fundus photography.The patient managed with intravitreal anti-vascular endothelial growth factor(anti-VEGF)injections(bevacizumab 1.25 mg/0.05 m L).The Cochrane Library,Pub Med,OVID,and Up To Date databases were searched using the term pseudoxanthoma elasticum or Gr?nblad-Strandberg syndrome with the limits English.Articles that predated the databases were gathered from current references.Fundus examination revealed angioid streaks bilaterally and CNV in left eye(LE).After the patient underwent three intravitreal injections of bevacizumab,the LE showed absorption of the subretinal fluid and shrinkage of the CNV.Visual acuity(VA)was improved in her treated LE.Bevacizumab treatment was well tolerated with no adverse events reported.Approximately ten articles about 45 patients(49 eyes)describing CNV secondary to angioid streaks in PXE treated with anti-VEGF were found in the literature search.In the present case,bevacizumab of an initial three injection loading dose,achieved maintenance of visual function in the treatment of CNV associated with angioid streaks in PXE.Literature articles concluded that the intravitreal application of anti-VEGF is highly efficient for improving and stabilizing the lesion as well as the eyesight.So we believe that anti-VEGF therapy can be a great choice of treatment for CNV secondary to angioid streaks related PXE.展开更多
AIM:To quantify and compare longitudinal thickness changes of the ganglion cell complex(GCC)and the choroid in patients with different patterns of age-related macular degeneration(AMD)progression.METHODS:Retrospective...AIM:To quantify and compare longitudinal thickness changes of the ganglion cell complex(GCC)and the choroid in patients with different patterns of age-related macular degeneration(AMD)progression.METHODS:Retrospective cohort analysis of anonymized data from participants aged 50y or more and diagnosed with early/intermediate AMD in at least one eye(with no evidence of advanced AMD).A total of 64 participants were included from the Instituto de Retina de Lisboa(IRL)study(IPL/2022/MetAllAMD_ESTeSL)and divided into 4 groups according to the Rotterdam classification for AMD.Spectral domain optical coherence tomography(SD-OCT)was used to assess and quantify GCC and choroid thickness at two time points(first visit vs last visit)with a minimum interval of 3y.RESULTS:In the GCC inner ring,a thinner thickness(P=0.001)was observed in the atrophic AMD group(51.3±21.4μm)compared to the early AMD(84.3±11.5μm),intermediate AMD(77.6±16.1μm)and neovascular AMD(88.9±16.3μm)groups.Choroidal thickness quantification showed a generalized reduction in the central circle(P=0.002)and inner ring(P=0.001).Slight reductions in retinal thickness were more accentuated in the inner ring in the atrophic AMD(-13%;P<0.01).CONCLUSION:The variation of the analyzed structures could be an indicator of risk of progression with neurodegenerative(GCC)or vascular(choroid)pattern in the intermediate and atrophic AMD.The quantification of both structures can provide important information about the risk of disease progression in the early and intermediate stages but also for the evolution pattern into late stages(atrophic or neovascular).展开更多
AIM: To evaluate the effects of lentivirus-mediated pigment epithelium-derived factor (PEDF) gene transfer performed in treatment of rats with established choroidal neovascularization (CNV), and investigates the mecha...AIM: To evaluate the effects of lentivirus-mediated pigment epithelium-derived factor (PEDF) gene transfer performed in treatment of rats with established choroidal neovascularization (CNV), and investigates the mechanism by which PEDF inhibits CNV in rats. METHODS: Brown Norway (BN) rats (n=204) were induced by exposure to a laser, and then randomly assigned to 3 groups: no treatment; treatments with intravitreal injection of lentivirus-PEDF-green fluorescent protein (GFP) or lentivirus-control GFP (free fluorescent protein). Following induction and treatment, the CNV tissue was assessed for form, size and vessel leakage by fluorescein fundus angiography (FFA), optical coherence tomography (OCT), histopathology, and examination of choroidal flat mounts. VEGF, Flk-1, and PEDF expression were evaluated by real-time polymerase chain reaction (PCR) and Western blot. RESULTS: A stable laser-induced rat model of CNV was successfully established, and used to demonstrate lentivirus-mediated REDO gene transfer by intravitreal injection. Expression of green fluorescence labelled PEDF was observed in the retina up to 28d after injection. An intravitreal injection of lentivirus-PEDF-GFP at 7d led to a significant reduction in the size, thickness and area of CNV showed by FFA, OCT and choroidal flat mounts. PEDF was up-regulated while VEGF and Flk-1 were down-regulated in the lentivirus-PEDF-GFP group. The differences in VEGF and Flk-1 expression in the control and lentivirus-PEDF groups at 7, 14, 21 and 28d after laser induction were all statistically significant. CONCLUSION: Lentivirus-mediated PEDF gene transfer is effective for use in treatment of laser-induced CNV, and PEDF exerts its therapeutic effects by inhibiting expression of VEGF and Flk-1.展开更多
AIM: To investigate the clinical and optical coherence tomography(OCT) features of focal choroidal excavation(FCE) complicated with choroidal neovascularization(CNV) in young and middle aged patients. METHODS: We perf...AIM: To investigate the clinical and optical coherence tomography(OCT) features of focal choroidal excavation(FCE) complicated with choroidal neovascularization(CNV) in young and middle aged patients. METHODS: We performed a retrospective review of 26 patients with FCE accompanied by CNV. All patients underwent a complete ophthalmic examination. We analyzed the clinical characteristics of patients, focusing on the spectral-domain OCT features. All patients received intravitreal injection of anti-vascular endothelial growth factor(anti-VEGF) agents. And we assessed the changes of central retinal thickness and best-corrected visual acuity(BCVA) after anti-VEGF therapy. RESULTS: The mean age of 26 patients was 35.5±7.3 y(range, 21-48 y). Of the 26 FCE lesions, 11 were located subfoveal, 6 were parafoveal, and 9 were extrafoveal. The mean FCE depth was 129.8±50.3 μm, and the mean width was 901.3±306.0 μm. The FCE depth was correlated positively with the width, but not correlated with age or refractive error. CNV was located within the excavation(19 eyes) or adjacent to the excavation(7 eyes). After anti-VEGF therapy, the central retinal thickness was significantly reduced and the BCVA was significantly improved. In the absorption process of subretinal fluid, we found that the fluid in the excavations needed to be absorbed at the last. A small amount of residual fluid could still be seen in a few deep excavations even after a longterm follow-up.CONCLUSION: FCE may be an important reason to cause CNV. Especially in young patients with idiopathic CNV, we should pay attention to the use of OCT to check the presence of FCE. Anti-VEGF therapy is generally effective for CNV associated with FCE.展开更多
Sonic hedgehog (Shh) signaling has recently been shown to be involved in the pathological angiogenesis in response to tissue hypoxia and ischemic injury.Hypoxia/ischemia is considered to play an important role in the ...Sonic hedgehog (Shh) signaling has recently been shown to be involved in the pathological angiogenesis in response to tissue hypoxia and ischemic injury.Hypoxia/ischemia is considered to play an important role in the development of choroidal neovascularization (CNV).This study was aimed to examine the effect of blockade of the Shh signaling pathway on CNV and the underlying mechanism.A total of 64 male Brown-Norway (BN) rats were used in this study.One eye of each rat underwent laser photocoagulation.The other eye served as normal control.After the laser treatment, the 64 rats were divided into four groups (n=16 in each group):Blank control group, in which no intravitreal administration was given; cyclopamine group, recombinant Shh N-terminals protein (rShh) group and phosphate-buffered saline (PBS) group, in which cyclopamine (a Shh inhibitor), rShh (a Shh activator) and PBS were intravitreally injected into the laser-treated eyes respectively every other day for a total of four intravitreal injections immediately after the laser treatment.Fourteen days after the intravitreal administration, the changes of CNV-related variables, including positive CNV lesion percentage, CNV membrane area and CNV membrane thickness, were evaluated by fluorescein anqiography, indocyanine green angiography and pathological examinations.The mRNA and protein expression of PTCH1, Gli1, HIF-1α, VEGF and DLL4 in each group on 14 days of CNV model was detected by real-time quantitative PCR and western blot analysis, and the relationship between the Shh cascade and the HIF-1α-VEGF-DLL4 cascade in CNV was analyzed.The results showed that the CNV membrane area and the CNV membrane thickness were decreased by 62.5% and 41.9% in the cyclopamine group and increased by 85.7% and 64.3% in the rShh group in comparison to those in the blank control group (P【0.01 for each).There was no significant difference in the CNV membrane area and thickness between the blank control group and PBS group (P=0.102 and P=0.063, respectively).Real-time quantitative PCR revealed a 5.23-, 4.14-, 2.97-, 2.78-and 2.39-fold up-regulation of the mRNA expression of PTCH1, Gli1, HIF-1α, VEGF and DLL4 genes in the laser-treated eyes compared with the normal control eyes in the control group.In the cyclopamine group, the mRNA and protein expression of Gli1, HIF-1α, VEGF and DLL4 was significantly down-regulated (P【0.05 for each) while the expression of PTCH1 showed no significant changes at the mRNA (P=0.293) and protein level (P=0.304).The mRNA expression and protein expression (P=0.001 and P=0.021, respectively) of PTCH1, Gli1, HIF-1α, VEGF and DLL4 was significantly increased in the rShh group when compared with the control group.The expression level of these genes was related to the severity of the CNV.It was concluded that intravitreal administration of cyclopamine can effectively inhibit the formation of laser-induced experimental CNV by down-regulating the expression of the HIF-1α-VEGF-DLL4 cascade in CNV.The Shh signaling pathway as an upstream signaling pathway of HIF-1α-VEGF-DLL4 cascade is implicated in the development of experimental CNV.展开更多
AIM:To evaluate the predictors of visual improvement in eyes with naive choroidal neovascularization secondary to age-related macular degeneration (CNV -AMD) treated with intravitreal bevacizumab (IVB) monotherapy. ME...AIM:To evaluate the predictors of visual improvement in eyes with naive choroidal neovascularization secondary to age-related macular degeneration (CNV -AMD) treated with intravitreal bevacizumab (IVB) monotherapy. METHODS:Fifty eyes with naive CNV-AMD with pretreatment best-corrected visual acuity (BCVA) better than 20/200 and treated with IVB monotherapy were evaluated. Several variables including age, sex, pre-treatment BCVA, CNV type and lesion size on fluorescein angiogram as well as SD-OCT parameters including pre-treatment central macular thickness (CMT), inner-segment/outer-segment (IS/OS) junction integrity, and external limiting membrane (ELM) integrity were analyzed to predict visual outcome.RESULTS:On univariate regression, pretreatment ELM damage was associated with less visual improvement after treatment (P =0.0145). However, ELM damage predicted only 10% of the visual outcome. On multivariate regression, pretreatment BCVA, IS/OS junction, and ELM integrity on SD-OCT were the significant predictors for the treatment effect and together predicted 37% of visual improvement. CONCLUSION:Pretreatment BCVA, ELM and IS/OS junction integrity on SD-OCT are of significant value inpredicting the visual improvement in naive wet AMD patients treated with IVB monotherapy.展开更多
Pigment epithelium derived factor (PEDF) has been proven to be an effective drug for the treatment of choroidal neovascularization (CNV).However,the lack of ideal administration route is the biggest bottleneck prevent...Pigment epithelium derived factor (PEDF) has been proven to be an effective drug for the treatment of choroidal neovascularization (CNV).However,the lack of ideal administration route is the biggest bottleneck preventing PEDF from wider clinical use.In this study,we developed a novel PEDF-carrying system which employed immuno-nano-liposomes (INLs) under ultrasound exposure.PEDF-loaded INLs were prepared by conjugating nanoliposomes to the peptide ATWLPPR specifically targeting the receptor-2 for vascular endothelial growth factor (VEGFR-2) and reversely encapsuling PEDF.RF/6A cells were incubated with PEDF-loaded INLs.CNV models of BN rats were injected with PEDF-loaded INLs.MTT assay was used to evaluate the cytotoxicity of the INLs on RF/6A cells.Flow cytometry was conducted to detect the apoptotic rate of cells.Laser scanning confocal microscopy was employed to observe the binding and transmitting process of PEDF-loaded INLs and to calculate the area of CNV in the rat model.The results showed that the PEDF-loaded INLs could exclusively bind to CNV but not to the normal choroidal vessels.The CNV area was significantly decreased in PEDF treatment groups in comparison with control group (P【0.05).Moreover,PEDF-loaded INLs exposed under ultrasound were more efficient in reducing the CNV area (P【0.05).It was concluded that INLs in combination with ultrasonic exposure can transmit PEDF into cytoplasma with high specificity and efficiency,which strengthens the inhibitory effects of PEDF on CNV and reduces its side effects.PEDF-loaded INLs possibly represent a new treatment paradigm for patients with ocular neovascularization.展开更多
Purpose: Choroidal neovascularization (CNV) plays an important role in pathogenesis of age-related macular degeneration (AMD), ocular histoplasmosis syndrome (OHS) and so on. However, mechanisms of CNV formation are n...Purpose: Choroidal neovascularization (CNV) plays an important role in pathogenesis of age-related macular degeneration (AMD), ocular histoplasmosis syndrome (OHS) and so on. However, mechanisms of CNV formation are not fully understood. The aim of this study is to investigate the correlation between expressions of CD105 and vascular endothelial growth factor (VEGF) in experimental laser-induced CNV in rats. Methods: CNV model was established by 532 nm laser photocoagulation in Brown-Norway rats. The expression of CD105 and VEGF in CNV was observed by immunohistochemistry at 3, 7, 14, 21, 28 and 56 days after laser photocoagulation. The image analysis was performed with the professional software of Image-Pro Plus. Results: Fluorescein angiography showed fluorescein leakage in CNV from days 7 to 56 after photocoagulation. VEGF expression was mainly observed in vascular endothelial cells, ganglion cells, inner nuclear layers and retinal pigment epithelial cells in normal retina and vascular endothelial cells in normal choroid of the rats. On day 3 after photocoagulation, VEGF began to express in laser-induced lesions. VEGF was strongly expressed in CNV after 7 days (P<0.05) and decreased after 14 days (P>0.05). CD105 was initially presented in CNV at 7 days and obviously expressed at 14 days after photocoagulation (P<0.05). Four weeks later, when angiogenesis tended toward inactive status, expression of CD105 was markedly decreased (P>0.05). There was notablely direct correlation between CD105-positive-microvessel density and positively semiquantitative scoring of VEGF in the CNV(r=0.989, P<0.01). Conclusions: There is direct correlation between the expression of CD105 and VEGF in the laser-induced CNV in rat. It suggests that CD105 and VEGF might participate in the new blood vessel formation and promote the growth of CNV.展开更多
Choroidal neovascularization(CNV) is a leading cause of visual loss in age-related macular degeneration(AMD). However, the molecular mechanism for CNV progression is still unclear. This study aimed to identify CNV-rel...Choroidal neovascularization(CNV) is a leading cause of visual loss in age-related macular degeneration(AMD). However, the molecular mechanism for CNV progression is still unclear. This study aimed to identify CNV-related circular RNAs(circRNAs), a novel class of non-coding RNAs with diverse functions. A total of 117 circRNAs were differentially expressed in the murine CNV model by microarrays. Gene ontology(GO)enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) analysis were performed to identify the functions of selected circRNAs. The host genes of these circRNAs were predicted to be targeted to neurogenesis(ontology: biological process), proteinaceous extracellular matrix(ECM)(ontology: cellular component), and binding(ontology: molecular function). Differentially expressed circRNAs-mediated regulatory networks were enriched in ECM receptor interaction. Most of the dysregulated circRNAs could potentially bind to five different mi RNAs by Target Scan and mi Randa. Specifically, circ_15752 was identified in this circRNAs pool which may facilitate vascular endothelial cell proliferation, migration, and tube formation, suggesting a critical role in endothelial angiogenesis. Our work suggests that dysregulated circRNAs may be involved in CNV pathogenesis and serve as potential biomarkers for CNV.展开更多
AIM:To evaluate whether recombinant complement factor B(CFB)short hairpin RNA(sh RNA)reduces laserinduced choroidal neovascularization(CNV)in rats.METHODS:Laser-induced rat CNV model was established,and then the anima...AIM:To evaluate whether recombinant complement factor B(CFB)short hairpin RNA(sh RNA)reduces laserinduced choroidal neovascularization(CNV)in rats.METHODS:Laser-induced rat CNV model was established,and then the animals underwent fundus fluorescence angiography(FFA)and hematoxylin and eosin(HE)staining.On day 3 and 7 after photocoagulation,the expression of CFB and membrane attack complex(MAC)was detected by immunhischemistry.A recombinant CFBsh RNA plasmid was constructed.CFB and scrambled sh RNA plasmids were intravenous injected into rats via the tail vein on the day of laser treatment,respectively.On day 7,the incidence of CNV was determined by FFA,and the expression of CFB and vascular endothelial growth factor(VEGF)in retinal pigment epithelium(RPE)/choroidal tissues was detected by immunhischemistry,Western blot and/or semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR)in CFB and scrambled sh RNA groups.The possible adverse effects of CFB-sh RNA injection were assessed by transmission electron microscopy and electroretinography.RESULTS:FFA and HE results indicated that a laserinduced rat CNV model was successfully established on day 7 after photocoagulation.The expression of CFB and MAC was extremely weak in normal retina and choroid,and increased on day 3 after photocoagulation.However,it started to reduce on day 7.CFB sh RNA plasmid was successfully constructed and induced CFB knockdown in the retinal and choroidal tissues.FFA showed CFB knockdown significantly inhibited incidence of CNV in rats.Moreover,CFB knockdown significantly inhibited the expression of VEGF in RPE/choroidal tissues.CFB sh RNA caused no obvious side effects in eyes.CONCLUSION:CFB knockdown significantly inhibits the formation and development of CNV in vivo through reducing the expression of VEGF,which is a potential therapy target.The alternative pathway of complement activation plays an important role in CNV formation.展开更多
AIM: To examine the effect of intravitreal adenoviral vector-mediated tristetraprolin(Ad-TTP) on VEGF m RNA expression in a rat model of laser-induced choroidal neovascularization.METHODS: Ad-TTP was prepared using a ...AIM: To examine the effect of intravitreal adenoviral vector-mediated tristetraprolin(Ad-TTP) on VEGF m RNA expression in a rat model of laser-induced choroidal neovascularization.METHODS: Ad-TTP was prepared using a commercial kit. Retinal laser-induced photocoagulation(10 spots per eye) was performed on rats in this experimental choroidal neovascularization(CNV) model. Rats were divided into four groups: control(single intravitreal injection of balanced salt solution, n =10), laser-induced CNV(photocoagulation only, n =20), laser-induced CNV plus Ad-TTP injection(photocoagulation plus a single intravitreal Ad-TTP injection, n =20) and Ad-TTP injection only(n =10). Changes in choroidal morphology were evaluated in ten rats in the laser only and the laser plus Ad-TTP groups. Two weeks after laser injury, the size of CNV was calculated by perfusion with high-molecular-weight fluorescein isothiocyanate(FITC)-dextran. VEGF m RNA expression in retina-choroid tissue from ten rats in each group was measured by reverse transcription polymerase chain reaction(RT-PCR). RESULTS: Two weeks after treatment, the area of laser-induced CNV was reduced by approximately 60% in the rats given the Ad-TTP injection compared with that in the laser-only group. There was a tendency toward decreased VEGF m RNA expression in the Ad-TTP injection groups.CONCLUSION: A single intravitreal injection of Ad-TTP significantly suppressed CNV size in this experimental laser-induced CNV model. Ad-TTP injection also decreased VEGF m RNA expression compared with that inthe laser-induced CNV group. The present study is meaningful as the first study to investigate the effect of tristetraprolin delivered via intravitreal injection.展开更多
AIM:To investigate the roles of integrins in choroidal neovascularization(CNV) and their associations with the stromal cell-derived factor-1(SDF-1)/CXCR4 axis.METHODS:CNV lesions were induced in mice using laser...AIM:To investigate the roles of integrins in choroidal neovascularization(CNV) and their associations with the stromal cell-derived factor-1(SDF-1)/CXCR4 axis.METHODS:CNV lesions were induced in mice using laser photocoagulation.After CNV induction,all animals were randomly assigned to:control,SDF-1,SDF-1+age-related macular degeneration(AMD) 3100(CXCR4 inhibitor),and SDF-1+ATN161(integrin α5β1 inhibitor) groups;their effects on CNV progression were observed using hematoxylin eosin(HE) staining,fundus fluorescein angiography(FFA) grading and optical coherence tomography(OCT),and their effects on CXCR4/integrin α5 expression were evaluated using Western blot and double immunofluorescence staining.Hypoxia-exposed endothelial cells(ECs) were used to simulate CNV in vitro,they were treated with SDF-1,combined with CXCR4 siRNA/AMD3100 or ATN161,and expression of integrin α5,cell migration and tube formation were analyzed.RESULTS:Integrin subunit α5 increased at 3^ rd and 7^ th day and decreased at 14 ^th day in CNV mice,with no significant change of β1-integrin.CXCR4 expression in CNV mice had persistent increase within 14 d after induction.SDF-1 treatment significantly promoted the CNV progression during 3-14 d.The mean CNV length in AMD3100 andATN161 group at day 7 was 270.13 and 264.23 μm in HE images,significantly lower than the mean length in SDF-1(345.70 μm) group.AMD3100 and ATN161 also significantly reduced thickness and leakage of CNV induced by SDF-1.Mean integrin α5 positive area in SDF-1 group reached 2.31×104 μm^2,significantly higher than control(1.25×104 μm^2),which decreased to 1.78×104 μm^2 after AMD3100 treatment.About 61.36% of ECs in CNV lesions expressed α5 in SDF-1 group,which significantly decreased to 43.12% after AMD3100 treatment.In vitro,integrin α5 peaked by 6 folds after 6 h of hypoxia exposure and CXCR4 gradually increased by up to 2.3 folds after 24 h of hypoxia.Approximately 25.12% of ECs expressed integrin α5 after SDF-1 stimulation,which decreased to 7.2%-9.5% after si-CXCR4 or AMD3100 treatment.ATN161 exerted an inhibitory effect comparable to that of si-CXCR4 on EC migration and tube formation in the presence of SDF-1.CONCLUSION:SDF-1/CXCR4 signaling induces integrin α5β1 expression in ECs to promote CNV.展开更多
AIM:To determine the effects of intravitreal resveratrol(RSV)on murine laser-induced choroidal neovascularization(CNV).METHODS:The toxicity of RSV to choroidal endothelial cell(CEC)was measured using thiazolyl blue te...AIM:To determine the effects of intravitreal resveratrol(RSV)on murine laser-induced choroidal neovascularization(CNV).METHODS:The toxicity of RSV to choroidal endothelial cell(CEC)was measured using thiazolyl blue tetrazolium bromide(M一)assay.Effects of RSV on choroidal endothelial cell(CEC)migration were evaluated with a modified Boyden chamber assay,while tube formation was evaluated in a 2-D gel assay.CNV was induced by laser photocoagulation in mice.The effects of intravitreal injection of RSV on CNV development were evaluated by fluorescein angiography(FA),confocal analysis of isolectin B4 labeled choroidal flat mounts,and histologic examination of CNV membranes.Immunostaining was used to analyze the expression and phosphorylation of vascular endothelial growth factor receptor 2(VEGFR2).RESULTS:No significant cell toxicity was observed in CEC if the concentration of RSV was less than 200 pmol/L(P>0.05).RSV inhibited vascular endothelial growth factor(VEGF)-induced CEC migration(P<0.05)and tube formation(P<0.05)invitro.Furthermore,intravitrealinjectionof RSV significantly inhibited laser induced CNV formation in mice.The FA leakage,CNV volume and CNV area analysis revealed that there were 41%,45%,and 58%reduction in RSV-treated eyes(1.691±0.1032,178163±78623μm^3 and 6508±619.0μm^2,respectively)compared with those in control(2.724±0.08447,379676±98382μm3and16576±2646μm^2,respectively;P<0.05).Phospho-VEGFR2expression was much weaker in the sections of CNV lesions in RSV injected mice compared with that in control(P<0.05).CONCLUSION:Intravitreal injection of RSV exerts an inhibitory effect on CNV,which may through suppressing endothelial cell migration,tube formation and VEGFR2 phosphorylation.展开更多
AIM:To reveal whether and how Yes-associated protein(YAP)promotes the occurrence of subretinal fibrosis in agerelated macular degeneration(AMD).METHODS:Cobalt chloride(Co Cl2)was used in primary human umbilical vein e...AIM:To reveal whether and how Yes-associated protein(YAP)promotes the occurrence of subretinal fibrosis in agerelated macular degeneration(AMD).METHODS:Cobalt chloride(Co Cl2)was used in primary human umbilical vein endothelial cells(HUVECs)to induce hypoxia in vitro.Eight-week-old male C57 BL/6 J mice weighing 19-25 g were used for a choroidal neovascularization(CNV)model induced by laser photocoagulation in vivo.Expression levels of YAP,phosphorylated YAP,mesenchymal markers[αsmooth muscle actin(α-SMA),vimentin,and Snail],and endothelial cell markers(CD31 and zonula occludens 1)were measured by Western blotting,quantitative real-time PCR,and immunofluorescence microscopy.Small molecules YC-1(Lificiguat,a specific inhibitor of hypoxia-inducible factor 1α),CA3(CIL56,an inhibitor of YAP),and XMU-MP-1(an inhibitor of Hippo kinase MST1/2,which activates YAP)were used to explore the underlying mechanism.RESULTS:Co Cl2 increased expression of mesenchymal markers,decreased expression of endothelial cell markers,and enhanced the ability of primary HUVECs to proliferate and migrate.YC-1 suppressed hypoxia-induced endothelialto-mesenchymal transition(End MT).Moreover,hypoxia promoted total expression,inhibited phosphorylation,and enhanced the transcriptional activity of YAP.XMU-MP-1 enhanced hypoxia-induced End MT,whereas CA3 elicited the opposite effect.Expression of YAP,α-SMA,and vimentin were upregulated in the laser-induced CNV model.However,silencing of YAP by vitreous injection of small interfering RNA targeting YAP could reverse these changes.CONCLUSION:The findings reveal a critical role of the hypoxia-inducible factor-1α(HIF-1α)/YAP signaling axis in End MT and provide a new therapeutic target for treatment of subretinal fibrosis in AMD.展开更多
Dear Editor,I am Dr.Sabiha Gungor Kobat,from the Department of Ophthalmology,Elazig Health Sciences University,Elazig,Turkey.I am writing to present an exceedingly rare case of two siblings,one of whom developed Biett...Dear Editor,I am Dr.Sabiha Gungor Kobat,from the Department of Ophthalmology,Elazig Health Sciences University,Elazig,Turkey.I am writing to present an exceedingly rare case of two siblings,one of whom developed Bietti crystalline dystrophy(BCD)with choroidal neovascularization at the age of 13 years,and the other has asymptomatic BCD at 8 years old.展开更多
Choroidal neovascularization characterizes wet age-related macular degeneration.Choroidal neovascularization formation involves a primarily angiogenic process that is combined with both inflammation and proteolysis.A ...Choroidal neovascularization characterizes wet age-related macular degeneration.Choroidal neovascularization formation involves a primarily angiogenic process that is combined with both inflammation and proteolysis.A primary cause of choroidal neovascularization pathogenesis is alterations in pro-and anti-angiogenic factors derived from the retinal pigment epithelium,with vascular endothelium growth factor being mainly responsible for both clinical and experimental choroidal neovascularization.MicroRNAs(miRNAs)which are short,non-coding,endogenous RNA molecules have a major role in regulating various pathological processes,including inflammation and angiogenesis.A review of recent studies with the mouse laser-induced choroidal neovascularization model has shown alterations in miRNA expression in choroidal neovascularization tissues and could be potential therapeutic targets for wet age-related macular degeneration.Upregulation of miR-505(days 1 and 3 post-laser),miR-155(day 14)occurred in retina;miR-342-5p(days 3 and 7),miR-126-3p(day 14)in choroid;miR-23a,miR-24,miR-27a(day 7)in retina/choroid;miR-505(days 1 and 3)in retinal pigment epithelium/choroid;downregulation of miR-155(days 1 and 3),miR-29a,miR-29b,miR-29c(day 5),miR-93(day 14),miR-126(day 14)occurred in retinal pigment epithelium/choroid.Therapies using miRNA mimics or inhibitors were found to decrease choroidal neovascularization lesions.Choroidal neovascularization development was reduced by overexpression of miR-155,miR-188-5p,miR-(5,B,7),miR-126-3p,miR-342-5p,miR-93,miR-126,miR-195a-3p,miR-24,miR-21,miR-31,miR-150,and miR-184,or suppression of miR-505,miR-126-3p,miR-155,and miR-23/27.Further studies are warranted to determine miRNA expression in mouse laser-induced choroidal neovascularization models in order to validate and extend the reported findings.Important experimental variables need to be standardized;these include the strain and age of animals,gender,number and position of laser burns to the eye,laser parameters to induce choroidal neovascularization lesions including wavelength,power,spot size,and duration.展开更多
AIM:To evaluate celastrol's effect on choroidal neovascularization(CNV).METHODS:In this study,neovascular formation in vitro(tube formation and aortic ring culture)and in vivo(laser induced neovascular in mice)was...AIM:To evaluate celastrol's effect on choroidal neovascularization(CNV).METHODS:In this study,neovascular formation in vitro(tube formation and aortic ring culture)and in vivo(laser induced neovascular in mice)was treated with celastrol to evaluate this natural compound's impact on CNV.Western blot was applied to explore the possible mechanism for it.For in vitro assay,triplicate for each group was repeated at least three times.For in vivo assay,each group contains 5 mice.RESULTS:Celastrol supressed tube formation and aortic ring sprout neovascularization.In vitro assay exhibited that celastrol inhibiting vascular endothelial growth factor(VEGF)-induced proliferation and migration of human umbilical vein endothelial cells and human choroidal endothelial cells,and by blocking VEGF signaling.Furthermore,intraperitoneal administration of celastrol significantly reduced the area of laser-induced CNV in an in vivo mouse model.By day 14,the area of CNV had decreased by 49.15%and 80.26%in the 0.1 mg/kg celastrol-treated group(n=5)and in the 0.5 mg/kg celastrol treated group(n=5),respectively,compared to the vehicle-treated group(n=5).CONCLUSION:Celastrol inhibits CNV by inhibiting VEGF-induced proliferation and migration of vascular endothelial cells,indicating that celastrol is a potent,natural therapeutic compound for the prevention of CNV.展开更多
基金Supported by the National Natural Science Foundation of China(No.82171076,No.82301221)Shanghai Municipal Education Commission(No.2023ZKZD18).
文摘AIM:To investigate the effect ofβ-alanine(BA)on laserinduced choroidal neovascularization(CNV)mice models.METHODS:Laser-induced CNV mice models were established,and BA was administrated for one week and two weeks in advance,separately.Furthermore,retinal pigment epithelium(RPE)-choroid flat mounts were separated,and immunohistochemical staining was performed.The laser-induced CNV lesion areas were measured and compared.In addition,liver and kidney morphologies were observed to identify potential hepatorenal toxicity.RESULTS:Enlarged CNV lesion areas were observed in the BA treated group.No significant differences were observed in the liver and kidney sections between groups.CONCLUSION:BA treatment increase CNV lesion areas,suggesting the detrimental effects of BA as a nutritional supplement in age-related macular degeneration(AMD)population.
文摘AIM:To observe the long-term clinical efficacy of intravitreal injections of conbercept,a novel vascular growth factor inhibitor,for the treatment of pathological myopia choroidal neovascularization(PM-CNV).METHODS:A total of 67 eyes(from 67 patients;mean age,54.90±12.7y)with PM-CNV were retrospectively researched.Based on the different schemes used for the administration of the drug,the patients were divided into two groups:group A(n=35;average age,53.31±13.6y;average diopter,9.25±1.72 D),which received only one injection of pro re nata(PRN;1+PRN regimen),and group B(n=32;average age,56.49±11.8y;average diopter,9.63±2.24 D),which received one injection per month for 3mo(3+PRN regimen).Best-corrected visual acuity(BCVA)analysis,intraocular pressure(IOP)examination,slit-lamp microscopy,fundus examination and optical coherence tomography were per formed at each follow-up.The recurrence and treatment times of CNV were recorded.The patients were followed up for at least 12mo.RESULTS:The BCVA was increased in 29 eyes(82.9%)in group A and 30 eyes(93.75%)in group B;no increase or decrease was observed in 6(17.1%)and 2(6.25%)eyes in groups A and B,respectively.The BCVA(log MAR)values before treatment(0.67±0.48 and 0.71±0.56)were significantly higher than those 12mo after treatment(0.31±0.26 and 0.33±0.17)in groups A and B,respectively(P<0.05).The mean central macular thickness(CMT)values had significantly decreased from 346.49±65.99 and 360.10±82.31μm at baseline to 257.29±40.47 and 251.97±48.26μm in groups A and B,respectively,after 12mo of treatment.A total of 21 eyes in group A needed reinjection(60%;average number of injections,2.51±0.98);the corresponding values in group B were 6 eyes(18.75%;average number of injections,3.74±1.22).There were no adverse ocular and systemic complications during the treatment and follow-up.CONCLUSION:Intravitreal injection of conbercept with 1+PRN or 3+PRN improve the visual acuity,reduce macular edema and reduce the level of CMT in patients with PM-CNV.The 3+PRN regimen demonstrates a lower recurrence rate of CNV than the 1+PRN regimen,but requires more treatment.However,both treatment regimens demonstrate long-term safety and efficacy for the treatment of PM-CNV.
基金Supported by the Shaanxi Key Research and Development Program-General Project in the Field of Social Development (No.2017SF-140)。
文摘AIM:To investigate whether anti-placental growth factor(PGF) can inhibit subretinal fibrosis and whether this effect is mediated by the inhibitory effect of PGF on epithelial-mesenchymal transition(EMT) of retinal pigment epithelial(RPE) cells.METHODS:Subretinal fibrosis model was established in laser induced choroidal neovascularization(CNV) mice on day 21 after laser photocoagulation.Immunofluorescence staining(IFS) of cryosections and enzyme-linked immunosorbent assay(ELISA) were used to detect the expression of PGF.IFS of whole choroidal flat-mounts was used to detect the degree of subretinal fibrosis.IFS of cryosections and ELISA were used to detect the expression of EMT related indicators in subretinal fibrosis lesions.RESULTS:The expression of PGF protein in subretinal fibrosis lesions was significantly up-regulated(P<0.05),and mainly co-stained with pan-cytokeratin labeled RPE cells.Intravitreal injection of anti-PGF neutralizing antibody reduced the area of subretinal fibrosis and the ratio of fibrotic/angiogenic area significantly at the concentrations of 0.25,0.5,1.0,and 2.0 μg/μL(all P<0.05).The expression of E-cadherin in the local RPE cells decreased,while α-SMA increased significantly in subretinal fibrosis lesions,and the application of anti-PGF neutralizing antibody could reverse these changes(P<0.05).CONCLUSION:The expression of PGF is up-regulated in the lesion site of subretinal fibrosis and mainly expressed in RPE cells.Intravitreal injection of anti-PGF neutralizing antibody can significantly inhibit the degree of subretinal fibrosis in CNV mice,and this effect may be mediated by the inhibition of PGF on EMT of RPE cells.
文摘The present study reports a case of a patient with choroidal neovascularization(CNV)associated with pseudoxanthoma elasticum(PXE).We observed the functional and anatomical improvement of the patient treated with intravitreal vascular endothelial growth factor(VEGF)inhibitor bevacizumab.The study also systematically searched the database for similar cases to provide a literature review.Data concerning the clinical features,treatment strategies and outcomes were extracted and analyzed.Retrospective interventional case report and systematic literature review.A 56-year-old healthy Chinese woman with CNV secondary to PXE was reported.Examinations included best corrected visual acuity(BCVA),biomicroscopy,optical coherence tomography(OCT),fluorescein and indocyanine green angiography and digital fundus photography.The patient managed with intravitreal anti-vascular endothelial growth factor(anti-VEGF)injections(bevacizumab 1.25 mg/0.05 m L).The Cochrane Library,Pub Med,OVID,and Up To Date databases were searched using the term pseudoxanthoma elasticum or Gr?nblad-Strandberg syndrome with the limits English.Articles that predated the databases were gathered from current references.Fundus examination revealed angioid streaks bilaterally and CNV in left eye(LE).After the patient underwent three intravitreal injections of bevacizumab,the LE showed absorption of the subretinal fluid and shrinkage of the CNV.Visual acuity(VA)was improved in her treated LE.Bevacizumab treatment was well tolerated with no adverse events reported.Approximately ten articles about 45 patients(49 eyes)describing CNV secondary to angioid streaks in PXE treated with anti-VEGF were found in the literature search.In the present case,bevacizumab of an initial three injection loading dose,achieved maintenance of visual function in the treatment of CNV associated with angioid streaks in PXE.Literature articles concluded that the intravitreal application of anti-VEGF is highly efficient for improving and stabilizing the lesion as well as the eyesight.So we believe that anti-VEGF therapy can be a great choice of treatment for CNV secondary to angioid streaks related PXE.
基金Supported by FCT/MCTES UIDB/05608/2020(https://doi.org/10.54499/UIDB/05608/2020)UIDP/05608/2020(https://doi.org/10.54499/UIDP/05608/2020)+1 种基金IDI&CA grant IPL/2022/MetAllAMD_ESTeSL by H&TRC-Health&Technology Research Center,ESTeSL-Escola Superior de Tecnologia da Saúde,Instituto Politécnico de Lisboaby Retina Institute of Lisbon(IRL).
文摘AIM:To quantify and compare longitudinal thickness changes of the ganglion cell complex(GCC)and the choroid in patients with different patterns of age-related macular degeneration(AMD)progression.METHODS:Retrospective cohort analysis of anonymized data from participants aged 50y or more and diagnosed with early/intermediate AMD in at least one eye(with no evidence of advanced AMD).A total of 64 participants were included from the Instituto de Retina de Lisboa(IRL)study(IPL/2022/MetAllAMD_ESTeSL)and divided into 4 groups according to the Rotterdam classification for AMD.Spectral domain optical coherence tomography(SD-OCT)was used to assess and quantify GCC and choroid thickness at two time points(first visit vs last visit)with a minimum interval of 3y.RESULTS:In the GCC inner ring,a thinner thickness(P=0.001)was observed in the atrophic AMD group(51.3±21.4μm)compared to the early AMD(84.3±11.5μm),intermediate AMD(77.6±16.1μm)and neovascular AMD(88.9±16.3μm)groups.Choroidal thickness quantification showed a generalized reduction in the central circle(P=0.002)and inner ring(P=0.001).Slight reductions in retinal thickness were more accentuated in the inner ring in the atrophic AMD(-13%;P<0.01).CONCLUSION:The variation of the analyzed structures could be an indicator of risk of progression with neurodegenerative(GCC)or vascular(choroid)pattern in the intermediate and atrophic AMD.The quantification of both structures can provide important information about the risk of disease progression in the early and intermediate stages but also for the evolution pattern into late stages(atrophic or neovascular).
基金Supported by the National Natural Science Foundation of China(No.81070735)
文摘AIM: To evaluate the effects of lentivirus-mediated pigment epithelium-derived factor (PEDF) gene transfer performed in treatment of rats with established choroidal neovascularization (CNV), and investigates the mechanism by which PEDF inhibits CNV in rats. METHODS: Brown Norway (BN) rats (n=204) were induced by exposure to a laser, and then randomly assigned to 3 groups: no treatment; treatments with intravitreal injection of lentivirus-PEDF-green fluorescent protein (GFP) or lentivirus-control GFP (free fluorescent protein). Following induction and treatment, the CNV tissue was assessed for form, size and vessel leakage by fluorescein fundus angiography (FFA), optical coherence tomography (OCT), histopathology, and examination of choroidal flat mounts. VEGF, Flk-1, and PEDF expression were evaluated by real-time polymerase chain reaction (PCR) and Western blot. RESULTS: A stable laser-induced rat model of CNV was successfully established, and used to demonstrate lentivirus-mediated REDO gene transfer by intravitreal injection. Expression of green fluorescence labelled PEDF was observed in the retina up to 28d after injection. An intravitreal injection of lentivirus-PEDF-GFP at 7d led to a significant reduction in the size, thickness and area of CNV showed by FFA, OCT and choroidal flat mounts. PEDF was up-regulated while VEGF and Flk-1 were down-regulated in the lentivirus-PEDF-GFP group. The differences in VEGF and Flk-1 expression in the control and lentivirus-PEDF groups at 7, 14, 21 and 28d after laser induction were all statistically significant. CONCLUSION: Lentivirus-mediated PEDF gene transfer is effective for use in treatment of laser-induced CNV, and PEDF exerts its therapeutic effects by inhibiting expression of VEGF and Flk-1.
基金Supported by the National Natural Science Foundation for Young Scholar of China(No.81600739)the Shanghai Hospital Development Center(No.SHDC12016116)the Science and Technology Commission of Shanghai Municipality(No.16411953700)
文摘AIM: To investigate the clinical and optical coherence tomography(OCT) features of focal choroidal excavation(FCE) complicated with choroidal neovascularization(CNV) in young and middle aged patients. METHODS: We performed a retrospective review of 26 patients with FCE accompanied by CNV. All patients underwent a complete ophthalmic examination. We analyzed the clinical characteristics of patients, focusing on the spectral-domain OCT features. All patients received intravitreal injection of anti-vascular endothelial growth factor(anti-VEGF) agents. And we assessed the changes of central retinal thickness and best-corrected visual acuity(BCVA) after anti-VEGF therapy. RESULTS: The mean age of 26 patients was 35.5±7.3 y(range, 21-48 y). Of the 26 FCE lesions, 11 were located subfoveal, 6 were parafoveal, and 9 were extrafoveal. The mean FCE depth was 129.8±50.3 μm, and the mean width was 901.3±306.0 μm. The FCE depth was correlated positively with the width, but not correlated with age or refractive error. CNV was located within the excavation(19 eyes) or adjacent to the excavation(7 eyes). After anti-VEGF therapy, the central retinal thickness was significantly reduced and the BCVA was significantly improved. In the absorption process of subretinal fluid, we found that the fluid in the excavations needed to be absorbed at the last. A small amount of residual fluid could still be seen in a few deep excavations even after a longterm follow-up.CONCLUSION: FCE may be an important reason to cause CNV. Especially in young patients with idiopathic CNV, we should pay attention to the use of OCT to check the presence of FCE. Anti-VEGF therapy is generally effective for CNV associated with FCE.
基金supported by grants from the National Natural Sciences Foundation of China (No.30901640)Natural Sciences Foundation of Hubei Provincial Government (No.2008CDB155)Chenguang Program of Wuhan Science and Technology Bureau (No.200950431170)
文摘Sonic hedgehog (Shh) signaling has recently been shown to be involved in the pathological angiogenesis in response to tissue hypoxia and ischemic injury.Hypoxia/ischemia is considered to play an important role in the development of choroidal neovascularization (CNV).This study was aimed to examine the effect of blockade of the Shh signaling pathway on CNV and the underlying mechanism.A total of 64 male Brown-Norway (BN) rats were used in this study.One eye of each rat underwent laser photocoagulation.The other eye served as normal control.After the laser treatment, the 64 rats were divided into four groups (n=16 in each group):Blank control group, in which no intravitreal administration was given; cyclopamine group, recombinant Shh N-terminals protein (rShh) group and phosphate-buffered saline (PBS) group, in which cyclopamine (a Shh inhibitor), rShh (a Shh activator) and PBS were intravitreally injected into the laser-treated eyes respectively every other day for a total of four intravitreal injections immediately after the laser treatment.Fourteen days after the intravitreal administration, the changes of CNV-related variables, including positive CNV lesion percentage, CNV membrane area and CNV membrane thickness, were evaluated by fluorescein anqiography, indocyanine green angiography and pathological examinations.The mRNA and protein expression of PTCH1, Gli1, HIF-1α, VEGF and DLL4 in each group on 14 days of CNV model was detected by real-time quantitative PCR and western blot analysis, and the relationship between the Shh cascade and the HIF-1α-VEGF-DLL4 cascade in CNV was analyzed.The results showed that the CNV membrane area and the CNV membrane thickness were decreased by 62.5% and 41.9% in the cyclopamine group and increased by 85.7% and 64.3% in the rShh group in comparison to those in the blank control group (P【0.01 for each).There was no significant difference in the CNV membrane area and thickness between the blank control group and PBS group (P=0.102 and P=0.063, respectively).Real-time quantitative PCR revealed a 5.23-, 4.14-, 2.97-, 2.78-and 2.39-fold up-regulation of the mRNA expression of PTCH1, Gli1, HIF-1α, VEGF and DLL4 genes in the laser-treated eyes compared with the normal control eyes in the control group.In the cyclopamine group, the mRNA and protein expression of Gli1, HIF-1α, VEGF and DLL4 was significantly down-regulated (P【0.05 for each) while the expression of PTCH1 showed no significant changes at the mRNA (P=0.293) and protein level (P=0.304).The mRNA expression and protein expression (P=0.001 and P=0.021, respectively) of PTCH1, Gli1, HIF-1α, VEGF and DLL4 was significantly increased in the rShh group when compared with the control group.The expression level of these genes was related to the severity of the CNV.It was concluded that intravitreal administration of cyclopamine can effectively inhibit the formation of laser-induced experimental CNV by down-regulating the expression of the HIF-1α-VEGF-DLL4 cascade in CNV.The Shh signaling pathway as an upstream signaling pathway of HIF-1α-VEGF-DLL4 cascade is implicated in the development of experimental CNV.
基金Supported by an Unrestricted Research Fund to Jacobs Retina Center at Shiley Eye Center,University of California, San Diego (LC)NIH EY 020617(LC)NIH EYO 7366 (WRF)
文摘AIM:To evaluate the predictors of visual improvement in eyes with naive choroidal neovascularization secondary to age-related macular degeneration (CNV -AMD) treated with intravitreal bevacizumab (IVB) monotherapy. METHODS:Fifty eyes with naive CNV-AMD with pretreatment best-corrected visual acuity (BCVA) better than 20/200 and treated with IVB monotherapy were evaluated. Several variables including age, sex, pre-treatment BCVA, CNV type and lesion size on fluorescein angiogram as well as SD-OCT parameters including pre-treatment central macular thickness (CMT), inner-segment/outer-segment (IS/OS) junction integrity, and external limiting membrane (ELM) integrity were analyzed to predict visual outcome.RESULTS:On univariate regression, pretreatment ELM damage was associated with less visual improvement after treatment (P =0.0145). However, ELM damage predicted only 10% of the visual outcome. On multivariate regression, pretreatment BCVA, IS/OS junction, and ELM integrity on SD-OCT were the significant predictors for the treatment effect and together predicted 37% of visual improvement. CONCLUSION:Pretreatment BCVA, ELM and IS/OS junction integrity on SD-OCT are of significant value inpredicting the visual improvement in naive wet AMD patients treated with IVB monotherapy.
基金supported by a grant from National Natural Sciences Foundation of China(No.30600691,30811260207)
文摘Pigment epithelium derived factor (PEDF) has been proven to be an effective drug for the treatment of choroidal neovascularization (CNV).However,the lack of ideal administration route is the biggest bottleneck preventing PEDF from wider clinical use.In this study,we developed a novel PEDF-carrying system which employed immuno-nano-liposomes (INLs) under ultrasound exposure.PEDF-loaded INLs were prepared by conjugating nanoliposomes to the peptide ATWLPPR specifically targeting the receptor-2 for vascular endothelial growth factor (VEGFR-2) and reversely encapsuling PEDF.RF/6A cells were incubated with PEDF-loaded INLs.CNV models of BN rats were injected with PEDF-loaded INLs.MTT assay was used to evaluate the cytotoxicity of the INLs on RF/6A cells.Flow cytometry was conducted to detect the apoptotic rate of cells.Laser scanning confocal microscopy was employed to observe the binding and transmitting process of PEDF-loaded INLs and to calculate the area of CNV in the rat model.The results showed that the PEDF-loaded INLs could exclusively bind to CNV but not to the normal choroidal vessels.The CNV area was significantly decreased in PEDF treatment groups in comparison with control group (P【0.05).Moreover,PEDF-loaded INLs exposed under ultrasound were more efficient in reducing the CNV area (P【0.05).It was concluded that INLs in combination with ultrasonic exposure can transmit PEDF into cytoplasma with high specificity and efficiency,which strengthens the inhibitory effects of PEDF on CNV and reduces its side effects.PEDF-loaded INLs possibly represent a new treatment paradigm for patients with ocular neovascularization.
基金Supported by National Natural Science Foundation ( No.30371516) , the Scientific Research Foundation for the ReturnedOverseas Chinese Scholars, State Education Ministry, China ( 2004) , Natural Science Foundation of Shanxi Province ( No.2004C246) and Scientific and Technological Innovation Foundation of Xijing Hospital ( No.XJCX04M003)
文摘Purpose: Choroidal neovascularization (CNV) plays an important role in pathogenesis of age-related macular degeneration (AMD), ocular histoplasmosis syndrome (OHS) and so on. However, mechanisms of CNV formation are not fully understood. The aim of this study is to investigate the correlation between expressions of CD105 and vascular endothelial growth factor (VEGF) in experimental laser-induced CNV in rats. Methods: CNV model was established by 532 nm laser photocoagulation in Brown-Norway rats. The expression of CD105 and VEGF in CNV was observed by immunohistochemistry at 3, 7, 14, 21, 28 and 56 days after laser photocoagulation. The image analysis was performed with the professional software of Image-Pro Plus. Results: Fluorescein angiography showed fluorescein leakage in CNV from days 7 to 56 after photocoagulation. VEGF expression was mainly observed in vascular endothelial cells, ganglion cells, inner nuclear layers and retinal pigment epithelial cells in normal retina and vascular endothelial cells in normal choroid of the rats. On day 3 after photocoagulation, VEGF began to express in laser-induced lesions. VEGF was strongly expressed in CNV after 7 days (P<0.05) and decreased after 14 days (P>0.05). CD105 was initially presented in CNV at 7 days and obviously expressed at 14 days after photocoagulation (P<0.05). Four weeks later, when angiogenesis tended toward inactive status, expression of CD105 was markedly decreased (P>0.05). There was notablely direct correlation between CD105-positive-microvessel density and positively semiquantitative scoring of VEGF in the CNV(r=0.989, P<0.01). Conclusions: There is direct correlation between the expression of CD105 and VEGF in the laser-induced CNV in rat. It suggests that CD105 and VEGF might participate in the new blood vessel formation and promote the growth of CNV.
基金supported by the National Natural Science Foundation of China(No.81525006,81670864,and 81730025 to C.Z.)the Non-profit Central Research Institute Fund of Chinese Academy of Medical Sciences(No.2018PT32019 to C.Z.)。
文摘Choroidal neovascularization(CNV) is a leading cause of visual loss in age-related macular degeneration(AMD). However, the molecular mechanism for CNV progression is still unclear. This study aimed to identify CNV-related circular RNAs(circRNAs), a novel class of non-coding RNAs with diverse functions. A total of 117 circRNAs were differentially expressed in the murine CNV model by microarrays. Gene ontology(GO)enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) analysis were performed to identify the functions of selected circRNAs. The host genes of these circRNAs were predicted to be targeted to neurogenesis(ontology: biological process), proteinaceous extracellular matrix(ECM)(ontology: cellular component), and binding(ontology: molecular function). Differentially expressed circRNAs-mediated regulatory networks were enriched in ECM receptor interaction. Most of the dysregulated circRNAs could potentially bind to five different mi RNAs by Target Scan and mi Randa. Specifically, circ_15752 was identified in this circRNAs pool which may facilitate vascular endothelial cell proliferation, migration, and tube formation, suggesting a critical role in endothelial angiogenesis. Our work suggests that dysregulated circRNAs may be involved in CNV pathogenesis and serve as potential biomarkers for CNV.
文摘AIM:To evaluate whether recombinant complement factor B(CFB)short hairpin RNA(sh RNA)reduces laserinduced choroidal neovascularization(CNV)in rats.METHODS:Laser-induced rat CNV model was established,and then the animals underwent fundus fluorescence angiography(FFA)and hematoxylin and eosin(HE)staining.On day 3 and 7 after photocoagulation,the expression of CFB and membrane attack complex(MAC)was detected by immunhischemistry.A recombinant CFBsh RNA plasmid was constructed.CFB and scrambled sh RNA plasmids were intravenous injected into rats via the tail vein on the day of laser treatment,respectively.On day 7,the incidence of CNV was determined by FFA,and the expression of CFB and vascular endothelial growth factor(VEGF)in retinal pigment epithelium(RPE)/choroidal tissues was detected by immunhischemistry,Western blot and/or semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR)in CFB and scrambled sh RNA groups.The possible adverse effects of CFB-sh RNA injection were assessed by transmission electron microscopy and electroretinography.RESULTS:FFA and HE results indicated that a laserinduced rat CNV model was successfully established on day 7 after photocoagulation.The expression of CFB and MAC was extremely weak in normal retina and choroid,and increased on day 3 after photocoagulation.However,it started to reduce on day 7.CFB sh RNA plasmid was successfully constructed and induced CFB knockdown in the retinal and choroidal tissues.FFA showed CFB knockdown significantly inhibited incidence of CNV in rats.Moreover,CFB knockdown significantly inhibited the expression of VEGF in RPE/choroidal tissues.CFB sh RNA caused no obvious side effects in eyes.CONCLUSION:CFB knockdown significantly inhibits the formation and development of CNV in vivo through reducing the expression of VEGF,which is a potential therapy target.The alternative pathway of complement activation plays an important role in CNV formation.
基金Supported by Biomedical Research Institute Fund (GNUHBRIF-2013-0002) from the Gyeongsang National University Hospital
文摘AIM: To examine the effect of intravitreal adenoviral vector-mediated tristetraprolin(Ad-TTP) on VEGF m RNA expression in a rat model of laser-induced choroidal neovascularization.METHODS: Ad-TTP was prepared using a commercial kit. Retinal laser-induced photocoagulation(10 spots per eye) was performed on rats in this experimental choroidal neovascularization(CNV) model. Rats were divided into four groups: control(single intravitreal injection of balanced salt solution, n =10), laser-induced CNV(photocoagulation only, n =20), laser-induced CNV plus Ad-TTP injection(photocoagulation plus a single intravitreal Ad-TTP injection, n =20) and Ad-TTP injection only(n =10). Changes in choroidal morphology were evaluated in ten rats in the laser only and the laser plus Ad-TTP groups. Two weeks after laser injury, the size of CNV was calculated by perfusion with high-molecular-weight fluorescein isothiocyanate(FITC)-dextran. VEGF m RNA expression in retina-choroid tissue from ten rats in each group was measured by reverse transcription polymerase chain reaction(RT-PCR). RESULTS: Two weeks after treatment, the area of laser-induced CNV was reduced by approximately 60% in the rats given the Ad-TTP injection compared with that in the laser-only group. There was a tendency toward decreased VEGF m RNA expression in the Ad-TTP injection groups.CONCLUSION: A single intravitreal injection of Ad-TTP significantly suppressed CNV size in this experimental laser-induced CNV model. Ad-TTP injection also decreased VEGF m RNA expression compared with that inthe laser-induced CNV group. The present study is meaningful as the first study to investigate the effect of tristetraprolin delivered via intravitreal injection.
基金Supported by the National Natural Science Foundation of China(No.81770936No.81570856+2 种基金No.81670863No.81500748No.81370020)
文摘AIM:To investigate the roles of integrins in choroidal neovascularization(CNV) and their associations with the stromal cell-derived factor-1(SDF-1)/CXCR4 axis.METHODS:CNV lesions were induced in mice using laser photocoagulation.After CNV induction,all animals were randomly assigned to:control,SDF-1,SDF-1+age-related macular degeneration(AMD) 3100(CXCR4 inhibitor),and SDF-1+ATN161(integrin α5β1 inhibitor) groups;their effects on CNV progression were observed using hematoxylin eosin(HE) staining,fundus fluorescein angiography(FFA) grading and optical coherence tomography(OCT),and their effects on CXCR4/integrin α5 expression were evaluated using Western blot and double immunofluorescence staining.Hypoxia-exposed endothelial cells(ECs) were used to simulate CNV in vitro,they were treated with SDF-1,combined with CXCR4 siRNA/AMD3100 or ATN161,and expression of integrin α5,cell migration and tube formation were analyzed.RESULTS:Integrin subunit α5 increased at 3^ rd and 7^ th day and decreased at 14 ^th day in CNV mice,with no significant change of β1-integrin.CXCR4 expression in CNV mice had persistent increase within 14 d after induction.SDF-1 treatment significantly promoted the CNV progression during 3-14 d.The mean CNV length in AMD3100 andATN161 group at day 7 was 270.13 and 264.23 μm in HE images,significantly lower than the mean length in SDF-1(345.70 μm) group.AMD3100 and ATN161 also significantly reduced thickness and leakage of CNV induced by SDF-1.Mean integrin α5 positive area in SDF-1 group reached 2.31×104 μm^2,significantly higher than control(1.25×104 μm^2),which decreased to 1.78×104 μm^2 after AMD3100 treatment.About 61.36% of ECs in CNV lesions expressed α5 in SDF-1 group,which significantly decreased to 43.12% after AMD3100 treatment.In vitro,integrin α5 peaked by 6 folds after 6 h of hypoxia exposure and CXCR4 gradually increased by up to 2.3 folds after 24 h of hypoxia.Approximately 25.12% of ECs expressed integrin α5 after SDF-1 stimulation,which decreased to 7.2%-9.5% after si-CXCR4 or AMD3100 treatment.ATN161 exerted an inhibitory effect comparable to that of si-CXCR4 on EC migration and tube formation in the presence of SDF-1.CONCLUSION:SDF-1/CXCR4 signaling induces integrin α5β1 expression in ECs to promote CNV.
基金Supported by the National Natural Science Foundation of China(No.81703134,No.81770952)Henan Province Nature Science Foundation of China(No.162300410296)Hunan Province Nature Science Foundation of China(No.2018JJ3772)。
文摘AIM:To determine the effects of intravitreal resveratrol(RSV)on murine laser-induced choroidal neovascularization(CNV).METHODS:The toxicity of RSV to choroidal endothelial cell(CEC)was measured using thiazolyl blue tetrazolium bromide(M一)assay.Effects of RSV on choroidal endothelial cell(CEC)migration were evaluated with a modified Boyden chamber assay,while tube formation was evaluated in a 2-D gel assay.CNV was induced by laser photocoagulation in mice.The effects of intravitreal injection of RSV on CNV development were evaluated by fluorescein angiography(FA),confocal analysis of isolectin B4 labeled choroidal flat mounts,and histologic examination of CNV membranes.Immunostaining was used to analyze the expression and phosphorylation of vascular endothelial growth factor receptor 2(VEGFR2).RESULTS:No significant cell toxicity was observed in CEC if the concentration of RSV was less than 200 pmol/L(P>0.05).RSV inhibited vascular endothelial growth factor(VEGF)-induced CEC migration(P<0.05)and tube formation(P<0.05)invitro.Furthermore,intravitrealinjectionof RSV significantly inhibited laser induced CNV formation in mice.The FA leakage,CNV volume and CNV area analysis revealed that there were 41%,45%,and 58%reduction in RSV-treated eyes(1.691±0.1032,178163±78623μm^3 and 6508±619.0μm^2,respectively)compared with those in control(2.724±0.08447,379676±98382μm3and16576±2646μm^2,respectively;P<0.05).Phospho-VEGFR2expression was much weaker in the sections of CNV lesions in RSV injected mice compared with that in control(P<0.05).CONCLUSION:Intravitreal injection of RSV exerts an inhibitory effect on CNV,which may through suppressing endothelial cell migration,tube formation and VEGFR2 phosphorylation.
基金National Natural Science Foundation of China(No.81970817,No.81873680)。
文摘AIM:To reveal whether and how Yes-associated protein(YAP)promotes the occurrence of subretinal fibrosis in agerelated macular degeneration(AMD).METHODS:Cobalt chloride(Co Cl2)was used in primary human umbilical vein endothelial cells(HUVECs)to induce hypoxia in vitro.Eight-week-old male C57 BL/6 J mice weighing 19-25 g were used for a choroidal neovascularization(CNV)model induced by laser photocoagulation in vivo.Expression levels of YAP,phosphorylated YAP,mesenchymal markers[αsmooth muscle actin(α-SMA),vimentin,and Snail],and endothelial cell markers(CD31 and zonula occludens 1)were measured by Western blotting,quantitative real-time PCR,and immunofluorescence microscopy.Small molecules YC-1(Lificiguat,a specific inhibitor of hypoxia-inducible factor 1α),CA3(CIL56,an inhibitor of YAP),and XMU-MP-1(an inhibitor of Hippo kinase MST1/2,which activates YAP)were used to explore the underlying mechanism.RESULTS:Co Cl2 increased expression of mesenchymal markers,decreased expression of endothelial cell markers,and enhanced the ability of primary HUVECs to proliferate and migrate.YC-1 suppressed hypoxia-induced endothelialto-mesenchymal transition(End MT).Moreover,hypoxia promoted total expression,inhibited phosphorylation,and enhanced the transcriptional activity of YAP.XMU-MP-1 enhanced hypoxia-induced End MT,whereas CA3 elicited the opposite effect.Expression of YAP,α-SMA,and vimentin were upregulated in the laser-induced CNV model.However,silencing of YAP by vitreous injection of small interfering RNA targeting YAP could reverse these changes.CONCLUSION:The findings reveal a critical role of the hypoxia-inducible factor-1α(HIF-1α)/YAP signaling axis in End MT and provide a new therapeutic target for treatment of subretinal fibrosis in AMD.
文摘Dear Editor,I am Dr.Sabiha Gungor Kobat,from the Department of Ophthalmology,Elazig Health Sciences University,Elazig,Turkey.I am writing to present an exceedingly rare case of two siblings,one of whom developed Bietti crystalline dystrophy(BCD)with choroidal neovascularization at the age of 13 years,and the other has asymptomatic BCD at 8 years old.
文摘Choroidal neovascularization characterizes wet age-related macular degeneration.Choroidal neovascularization formation involves a primarily angiogenic process that is combined with both inflammation and proteolysis.A primary cause of choroidal neovascularization pathogenesis is alterations in pro-and anti-angiogenic factors derived from the retinal pigment epithelium,with vascular endothelium growth factor being mainly responsible for both clinical and experimental choroidal neovascularization.MicroRNAs(miRNAs)which are short,non-coding,endogenous RNA molecules have a major role in regulating various pathological processes,including inflammation and angiogenesis.A review of recent studies with the mouse laser-induced choroidal neovascularization model has shown alterations in miRNA expression in choroidal neovascularization tissues and could be potential therapeutic targets for wet age-related macular degeneration.Upregulation of miR-505(days 1 and 3 post-laser),miR-155(day 14)occurred in retina;miR-342-5p(days 3 and 7),miR-126-3p(day 14)in choroid;miR-23a,miR-24,miR-27a(day 7)in retina/choroid;miR-505(days 1 and 3)in retinal pigment epithelium/choroid;downregulation of miR-155(days 1 and 3),miR-29a,miR-29b,miR-29c(day 5),miR-93(day 14),miR-126(day 14)occurred in retinal pigment epithelium/choroid.Therapies using miRNA mimics or inhibitors were found to decrease choroidal neovascularization lesions.Choroidal neovascularization development was reduced by overexpression of miR-155,miR-188-5p,miR-(5,B,7),miR-126-3p,miR-342-5p,miR-93,miR-126,miR-195a-3p,miR-24,miR-21,miR-31,miR-150,and miR-184,or suppression of miR-505,miR-126-3p,miR-155,and miR-23/27.Further studies are warranted to determine miRNA expression in mouse laser-induced choroidal neovascularization models in order to validate and extend the reported findings.Important experimental variables need to be standardized;these include the strain and age of animals,gender,number and position of laser burns to the eye,laser parameters to induce choroidal neovascularization lesions including wavelength,power,spot size,and duration.
基金Supported by National Natural Science Foundation of China(No.81570826)。
文摘AIM:To evaluate celastrol's effect on choroidal neovascularization(CNV).METHODS:In this study,neovascular formation in vitro(tube formation and aortic ring culture)and in vivo(laser induced neovascular in mice)was treated with celastrol to evaluate this natural compound's impact on CNV.Western blot was applied to explore the possible mechanism for it.For in vitro assay,triplicate for each group was repeated at least three times.For in vivo assay,each group contains 5 mice.RESULTS:Celastrol supressed tube formation and aortic ring sprout neovascularization.In vitro assay exhibited that celastrol inhibiting vascular endothelial growth factor(VEGF)-induced proliferation and migration of human umbilical vein endothelial cells and human choroidal endothelial cells,and by blocking VEGF signaling.Furthermore,intraperitoneal administration of celastrol significantly reduced the area of laser-induced CNV in an in vivo mouse model.By day 14,the area of CNV had decreased by 49.15%and 80.26%in the 0.1 mg/kg celastrol-treated group(n=5)and in the 0.5 mg/kg celastrol treated group(n=5),respectively,compared to the vehicle-treated group(n=5).CONCLUSION:Celastrol inhibits CNV by inhibiting VEGF-induced proliferation and migration of vascular endothelial cells,indicating that celastrol is a potent,natural therapeutic compound for the prevention of CNV.