Effects of AMD3100 subconjunctival injection on alkali burn induced corneal neovascularization in mice

AIM: To investigate the therapeutic effects of local and systemic administration of AMD3100 for alkali burn induced corneal neovascularization (CNV) in mice. METHODS: CNV was induced in vivo by alkaline burn of cornea in C57BL/6 mice. AMD3100 was administrated topically by subconjunctival injection or systemically by intraperitoneal injection for 7 days; balanced salt solution was administrated topically or systemically as a control respectively. Inflammatory index was evaluated by slit-lamp biomicroscopy and inflammatory cells infiltrated to cornea tissue were detected by histologic analysis at multiple time points. CNV was compared between the local and systemic treated mice 2 weeks after alkali burn, as quantified by CD34 immunostaining. Fluorescence-Activated Cell Sorter Analysis was used to investigate the mobilizing effects of EPC in mice after subconjunctival injected or intraperitoneal injected AMD3100. Immunohistochemistry was used to detect the expression of endothelial progenitor cells (EPC) marker proteins VEGFR2 and CD34. RESULTS: Three days after alkali burn, infiltration of inflammatory cells was found in corneal tissue. At the first 7 days of local injection group, the number of inflammatory cells was significantly lower than that in systemic injection group. CNV could be seen at the 7(th) day, and at the 14(th) day reached the peak, then started to decrease. The number of CNV in the subconjunctival injection group was 7.57 +/- 1.26 per 0.034mm(2), compared to a number of 14.87 +/- 2.21 per 0.034mm(2) in the control group (P<0.05). On the contrary, the number of CNV in the intraperitoneal injection group was a little higher than that in the control group, 16.34 +/- 1.53 per 0.034mm(2) vs 13.26 +/- 1.87 per 0.034mm(2). The research also showed that intraperitoneally, but not subconjunctivally injected AMD3100 could mobilize EPC. On the other hand, subconjunctival, but not intraperitoneally injected AMD3100 could reduce the expression of EPC marker proteins. CONCLUSION: In mice locally administrated AMD3100 can reduce the number of alkali burn induced CNV. The number of inflammatory cells and inflammatory responses in corneal tissue.

Effect of amnion membrane transplantation on corneal neovascularization in 10 patients with alkali burn

By observing clinical cases, we studied the curative effect of amnion membrane transplantation on decreasing corneal neovascularization (CNV). It was a non-randomized retrospective case-control study. Among 17 cases (21 eyes) of third-degree alkali burns from 2007 to 2010, 10 cases (12 eyes) were performed with amnion membrane transplantation operation, and others were not. Amnion membrane transplantation was performed at the 3rd day after burn in the treatment group. Areas of CNV in double groups were measured at the 14th day and 60th day after burn. Area of CNV in the treatment group was (66.207±7.251)mm2 at the 14th day after burn, and was 18.27% lower than that in the control group. Area of CNV in the treatment group was (120.046±13.812)mm2 at the 60th day after burn, and was 11.35% lower than that in the control group. There was both statistical significance (P<0.05). Amnion membrane transplantation operation can inhibit the growth of corneal neovascularization induced by alkali burn.

Inhibitory effect of polysulfated heparin endostatin on alkali burn induced corneal neovascularization in rabbits

AIM: To investigate anti-angiogenic effects of polysulfated heparin endostatin(PSH-ES) on alkali burn induced corneal neovascularization(NV) in rabbits.METHODS: An alkali burn was made on rabbit corneas to induce corneal NV in the right eye of 24 rabbits. One day after burn creation, a 0.2 m L subconjunctival injection of 50 μg/m L PSH-ES, 50 μg/m L recombinant endostatin(ES), or normal saline was administered every other day for a total of 14d(7 injections). Histology and immunohistochemisty were used to examine corneas.Corneal NV growth was evaluated as microvessel quantity and corneal vascular endothelial growth factor(VEGF)expression was measured by immunohistochemical assay.RESULTS: Subconjunctival injection of ES and PSHES resulted in significant corneal NV suppression, but PSH-ES had a more powerful anti-angiogenic effect than ES. Mean VEGF concentration in PSH-ES treated corneas was significantly lower than in ES treated and saline treated corneas. Histological examination showed that corneas treated with either PSH-ES or ES had significantly fewer microvessels than eyes treated with saline. Additionally corneas treated with PSH-ES had significantly fewer microvessels than corneas treated with ES.CONCLUSION: Both PSH-ES and recombinant ES effectively inhibit corneal NV induced by alkali burn.However, PSH-ES is a more powerful anti-angiogenic agent than ES. This research has the potential to provide a new treatment option for preventing and treating corneal NV.

Research on mouse model of grade Ⅱ corneal alkali burn

AIM： To choose appropriate concentration of sodium hydroxide（Na OH） solution to establish a stable and consistent corneal alkali burn mouse model in grade II.·METHODS： The mice（n =60） were randomly divided into four groups and 15 mice each group. Corneal alkali burns were induced by placing circle filter paper soaked with Na OH solutions on the right central cornea for 30 s.The concentrations of Na OH solutions of groups A, B, C,and D were 0.1 mol/L, 0.15 mol/L, 0.2 mol/L, and 1.0 mol/L respectively. Then these corneas were irrigated with 20 m L physiological saline（0.9% Na Cl）. On day 7 postburn, slit lamp microscope was used to observe corneal opacity,corneal epithelial sodium fluorescein staining positive rate, incidence of corneal ulcer and corneal neovascularization, meanwhile pictures of the anterior eyes were taken. Cirrus spectral domain optical coherence tomography was used to scan cornea to observe corneal epithelial defect and corneal ulcer.·RESULTS： Corneal opacity scores（ x ±s） were not significantly different between the group A and group B（P =0.097）. Incidence of corneal ulcer in group B was significantly higher than that in group A（P =0.035）.Incidence of corneal ulcer and perforation rate in group B was lower than that in group C. Groups C and D had corneal neovascularization, and incidence of corneal neovascularization in group D was significantly higher than that in group C（P =0.000）.·CONCLUSION： Using 0.15 mol/L Na OH can establish grade II mouse model of corneal alkali burns.

Experimental study on the treatment of rabbit corneal melting after alkali burn with Collagen cross-linking

AIM: To evaluate the effect of Collagen cross-linking on the prevention of melting in rabbit corneas after alkali burn. METHODS: Twenty New Zealand white rabbits were randomly divided into model control group and collagen cross-linking treatment group. The second group of rabbits received collagen cross linked treatment. Both groups were applied with antibiotic eye drops to prevent infection. The corneas were evaluated for melting, opacity, pathological and immunohistochemistry, record the changes when 28 days after the animals were killed. RESULTS: In the control group, 6 out of 8 rabbits showed corneal melting after injury (14 +/- 4) days, while two corneal perforated. In collagen cross-linking treatment group, one rabbit showed corneal melting after injury 23 days, without corneal perforation; corneal dissolution rate between the two groups was significantly different (P <0.05). Pathological examination suggested that in the treatment group, mild corneal edema, mild damage to collagen fibers, inflammatory cell infiltration was significantly less than the control group. Immunohistochemistry showed that corneal collagen fibers arranged in neat rows in the control group. CONCLUSION: Collagen cross-linking treatment not only can prevent and delay the corneal melting after alkali burn, but also can reduce the destruction of corneal collagen fibers and infiltration of inflammatory cells in the corneal tissue.

Efficacy of the nucleotide-binding oligomerization domain 1 inhibitor Nodinhibit-1 on corneal alkali burns in rats

AIM: To evaluate the therapeutic effect of Nodinhibit-1 on alkali-burn-induced corneal neovascularization (CNV) and inflammation. The nucleotide-binding oligomerzation domain 1 (NOD1) is a potent angiogenic gene. METHODS: The alkali -burned rat corneas (32 right eyes) were treated with eye drops containing Nodinhibit-1 or phosphate buffered solution (PBS, pH 7.4) only, four times per day. CNV and inflammation were monitored using slit lamp microscopy, and the area of CNV was measured by formula. Vascular endothelial growth factor (VEGF) and pigment epithelium -derived factor (PEDF) was determined by Western blot analysis. The TUNEL assay was used to assess the corneal apoptosis cells. RESULTS: Alkali-burn-induced progressive CNV and inflammation in the cornea. After treatment for 7d and 14d, there were statistically significant differences in the CNV areas and inflammatory index on that between two group(P<0.05, respectively). Epithelial defect quantification showed a significant difference between the two groups at days 4 and 7 after the alkali burns (P<0.05). The apoptotic. cells on days 1, 4, and 7 between the two groups showed significant differences at all time points (P<0.05, respectively). Compared to that in control group, the protein level of VEGF expression was significantly reduced whereas the PEDF expression was increase in the Nodinhibit-1 groups on day 14(P<0.05, respectively). CONCLUSION: Topical application of 10.0 mu g/mL Nodinhibit -1 may have potential effect for the alkali burn -induced CNV and inflammation. The effect of Nodinhibit -1 on CNV may be by regulation the equilibrium of VEGF and PEDF in the wounded cornea.

Effect of nintedanib thermo-sensitive hydrogel on neovascularization in alkali burn rat model

AIM:To investigate the effects of nintedanib thermo-sensitive hydrogel(NTH)on neovascularization and related markers in corneal alkali burns of Wistar rats.METHODS:NTH was prepared by grinding,and its phase-transition temperature was determined.Thirty specific-pathogen-free Wistar rats served as a model of corneal alkali burn in the right eye were randomly divided into 3 groups(n=10,each):model group treated with 0.9%saline once a day,NTH group with 0.2%nintedanib b.i.d,and dexamethasone group with dexamethasone ointment once a day.The left eye of rats served as the controls.The corneal transparency was observed under a slit-lamp microscope,and the area of neovascularization was calculated.On day 7,the rats were sacrificed,and the cornea was removed and embedded with paraffin,then stained with hematoxylin一eosin,and the expression of vascular endothelial growth factor receptor 2(VEGFR-2)and CD31 in the corneal tissues of each group was detected by immunofluorescence.RESULTS:The phase-transition temperature ofnintedanib obtained by grinding was 37℃after adding artificial tears.The results of the alkali burn model indicated that the growth rate of neovascularization in the NTH group was slower than that in the model group,and the neovascularization area was significantly smaller than that in the model group(P<0.05).Moreover,CD31 and VEGFR-2 expression levels in the NTH group were significantly lower than those in the model group.CONCLUSION:NTH becomes colloidal at body temperature,which is beneficial for releasing the drug slowly and can significantly inhibit the neovascularization of corneal induced by alkali burn in rats.

Effect of doxycycline intervention on the apoptosis as well as the IL-1, TNF-α and HIF-1α expression in cornea and aqueous humor in rats with corneal alkali burn

Objective:To study the effect of doxycycline intervention on the apoptosis as well as the IL-1, TNF-α and HIF-1α expression in cornea and aqueous humor in rats with corneal alkali burn.Methods: Male SD rats were selected as experimental animals and randomly divided into control group, alkali burn group and doxycycline group, corneal alkali burn models were established by sodium hydroxide eye drop and then they received doxycycline eye drops intervention. The expression of apoptosis molecules, inflammatory response cytokines and angiogenesis moleculesin the cornea as well as the expression of inflammatory response cytokines and angiogenesis molecules in aqueous humor were detected 14 and 28 d after model establishment.Results: 14 and 28 d after model establishment, Bcl-2 and PEDF protein expression in cornea tissue of alkali burn group were significantly lower than those of control group while Caspase-3, Caspase-8, Caspase-9, IL-1, TNF-α, HIF-1α and VEGF protein expression were significantly higher than those of control group;Bcl-2 and PEDF protein expression in cornea tissue of doxycycline group were significantly higher than those of alkali burn group while Caspase-3, Caspase-8, Caspase-9, IL-1, TNF-α, HIF-1α and VEGF protein expression were significantly lower than those of alkali burn group.Conclusion: Doxycycline for corneal alkali burn intervention can inhibit the apoptosis, inflammatory response and angiogenesis.

Inhibited experimental corneal neovascularization by neutralizing anti-SDF-1α antibody

AIM: To explore the effect of SDF-1α on the development of experimental corneal neovascularization (CRNV).METHODS: CRNV was induced by alkali injury in mice. The expression of SDF-1α and CXCR4 in burned corneas was examined by Flow Cytometry. Neutralizing anti-mouse SDF-1α antibody was locally administrated after alkali injury and the formation of CRNV 2 weeks after injury was assessed by Immunohistochemistry. The expression of VEGF and C-Kit in burned corneas was detected by RT-PCR.RESULTS: The number of CRNV peaks at 2 weeks after alkali injury. Compared to control group, SDF-1α neutralizing antibody treatment significantly decreased the number of CRNV. RT-PCR confirmed that SDF-1α neutralizing antibody treatment resulted in decreased intracorneal VEGF and C-Kit expression.CONCLUSION: SDF-1α neutralizing antibody treated mice exhibited impaired experimental CRNV through down regulated VEGF and C-Kit expression.

The Effect of Fibronectin on Re-epithelialization of Rabbits Cornea after Alkali Burn

The authors found experimentally that (1) fibronectin enhanced the healing of rabbit corneal epithelium after alkali burn and prevented the secondary breakdown; (2) it rapidly deposited on the denuded basement membrane to disappear as epithelial cells slided over, and (3) ultrastructurally, the neighbouring epithelial cells became flattened, with filopodia at the advancing edge, and extended to the wounded areas at 24 hours after the burn. However, the epithelial defects recurred 72 hours after the burn...

Emodin suppresses alkali burn-induced corneal inflammation and neovascularization by the vascular endothelial growth factor receptor 2 signaling pathway

OBJECTIVE:To investigate the effects of emodin on alkali burn-induced corneal inflammation and neovascularization.METHODS:The ability of emodin to target vascular endothelial growth factor receptor 2(VEGFR2)was predicted by molecular docking.The effects of emodin on the invasion,migration,and proliferation of human umbilical vein endothelial cells(HUVEC)were determined by cell counting kit-8,Transwell,and tube formation assays.Analysis of apoptosis was performed by flow cytometry.CD31 levels were examined by immunofluorescence.The abundance and phosphorylation state of VEGFR2,protein kinase B(Akt),signal transducer and activator of transcription 3(STAT3),and P38 were examined by immunoblot analysis.Corneal alkali burn was performed on 40 mice.Animals were divided randomly into two groups,and the alkali-burned eyes were then treated with drops of either 10μM emodin or phosphate buffered saline(PBS)four times a day.Slitlamp microscopy was used to evaluate inflammation and corneal neovascularization(CNV)in all eyes on Days 0,7,10,and 14.The mice were killed humanely 14 d after the alkali burn,and their corneas were removed and preserved at-80℃ until histological study or protein extraction.RESULTS:Molecular docking confirmed that emodin was able to target VEGFR2.The findings revealed that emodin decreased the invasion,migration,angiogenesis,and proliferation of HUVEC in a dose-dependent manner.In mice,emodin suppressed corneal inflammatory cell infiltration and inhibited the development of corneal neovascularization induced by alkali burn.Compared to those of the PBS-treated group,lower VEGFR2 expression and CD31 levels were found in the emodintreated group.Emodin dramatically decreased the expression of VEGFR2,p-VEGFR2,p-Akt,p-STAT3,and p-P38 in VEGF-treated HUVEC.CONCLUSION:This study provides a new avenue for evaluating the molecular mechanisms underlying corneal inflammation and neovascularization.Emodin might be a promising new therapeutic option for corneal alkali burns.

托珠单抗调控角膜碱烧伤后修复的研究

目的:评估外用白介素(Interleukin,IL)-6特异性抑制剂托珠单抗滴眼液在调控角膜碱烧伤后修复的安全性和有效性。方法:6只角膜假烧伤小鼠局部使用托珠单抗滴眼(2.5 mg/mL)和6只角膜假烧伤小鼠局部使用生理盐水滴眼,分别作为实验组和空白组以评估托珠单抗滴眼液的安全性。30只碱烧伤小鼠,按照1∶1随机分配到治疗组和对照组,治疗组使用托珠单抗滴眼液滴眼,对照组使用生理盐水,每日6次,连续用14 d。通过前段光学相干断层扫描观察虹膜前粘连、角膜后弹力层脱离及角膜水肿,在体视显微镜下检查角膜瘢痕形成及上皮伤口愈合。在角膜切片上评估IL-6定位、肌成纤维细胞、免疫细胞浸润和角膜上皮化生。在角膜铺片上评估角膜新生血管和新生淋巴管面积。通过实时荧光定量聚合酶链式反应(Quantitative Real-time Polymerase Chain Reaction,qRT-PCR)方法检测小鼠角膜IL-6的表达水平。结果:对未进行碱烧伤的角膜使用托珠单抗治疗未观察到明显的角膜结构的损伤。角膜碱烧伤后可见角膜结构的破坏,角膜瘢痕形成并伴有角膜上皮伤口的延迟愈合。使用托珠单抗治疗后,虹膜前粘连的发生率从86.67%下降至20%(P<0.01),角膜后弹力层脱离的发生率从93.33%下降至53.33%(P<0.05),角膜厚度小于对照组[(100.03±15.73)μm vs.(207.02±56.30)μm,P<0.001],角膜混浊评分从对照组的3.76±0.44下降到治疗组的1.94±0.83(P<0.001),治疗组在第5天(P<0.05)、第10天(P<0.001)和第14天(P<0.001)的上皮愈合率高于对照组。角膜碱烧伤后可见IL-6大量分布于角膜全层,且可见大量肌成纤维细胞形成及免疫细胞浸润,托珠单抗治疗后抑制了IL-6的表达(下降77.5%,P<0.05),肌成纤维细胞数量从每视野(91.44±65.60)个减少至(12.89±10.51)个(P<0.01),免疫细胞的数量从每视野(60.30±28.71)个细胞减少至每视野(6.80±3.82)个细胞(P<0.001)。此外,托珠单抗还减少角膜切片中每视野的杯状细胞数目,由(11.30±5.29)个减少至(2.00±1.90个)(P<0.01),并减少角膜新生血管和新生淋巴管的形成(分别减少了76.86%和71.16%,均P<0.001)。结论:局部使用托珠单抗抑制IL-6未见明显角膜毒性,且可以调控角膜碱烧伤后的修复。

枸杞糖肽对角膜碱烧伤小鼠角膜炎症反应及愈合的作用及其机制研究

目的探讨枸杞糖肽(LbGp)在角膜碱烧伤损伤愈合中的作用及其机制。方法根据治疗方式不同将45只健康、SPF级、6~8周、雄性C57BL/6J小鼠随机分为正常对照组(N组)、损伤对照组(PBS组)、LbGp治疗组(LbGp组),每组15只。PBS组和LbGp组小鼠右眼复制碱烧伤模型,采用点眼联合结膜下注射的方式分别给予PBS溶液或LbGp溶液(10 mg/mL)治疗。治疗后第3天采用裂隙灯显微镜及HE染色观察各组小鼠角膜上皮修复情况和角膜组织结构;实时荧光定量聚合酶链反应、Western blotting及免疫组织化学染色技术检测各组小鼠角膜组织NOD样受体热蛋白结构域相关蛋白3(NLRP3)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、髓过氧化物酶(MPO)的表达及核转录因子κB(NF-κB)的磷酸化水平。治疗后的第14天采用裂隙灯显微镜对角膜混浊程度进行评分,HE染色观察角膜组织结构。结果治疗后第3天LbGp组小鼠角膜上皮愈合速度快于PBS组(P<0.05)。N组、PBS组及LbGp组的NF-κB-p65、NLRP3、IL-1β及IL-6基因和蛋白相对表达量比较,差异均有统计学意义(P<0.05),PBS组均高于N组(P<0.05),LbGp组均低于PBS组(P<0.05)。LbGp组角膜基质中MPO染色阳性的中性粒细胞的浸润较PBS组减轻。治疗后第14天LbGp组小鼠角膜混浊评分低于PBS组(P<0.05)。结论LbGp治疗可以抑制小鼠角膜碱烧伤后NF-κB的激活及NLRP3、IL-1β的表达,从而减轻过度的炎症反应,促进小鼠角膜上皮再生及角膜结构恢复,有助于恢复角膜的透明性和完整性。

石斛酚抑制碱烧伤大鼠角膜新生血管的实验研究

目的 探讨石斛酚抑制碱烧伤大鼠角膜新生血管(corneal neovascularization, CNV)的作用。方法 构建SD大鼠角膜碱烧伤模型,随机分成正常对照组、模型对照组、低浓度石斛酚组、高浓度石斛酚组、阿柏西普组,每组各10只。低浓度石斛酚组、高浓度石斛酚组、阿柏西普组分别于伤后第1天结膜下注射2.5 mg/0.05 mL、5 mg/0.05 mL石斛酚,2 mg/0.05 mL阿柏西普。伤后第3、7、14天,观察并计算角膜新生血管占全角膜面积的百分比、角膜混浊评分并测量角膜厚度。碱烧伤第14天,处死全部大鼠,通过苏木素-伊红(HE)染色和免疫组化观察角膜组织中VEGF和CD34蛋白表达水平,以及通过ELISA检测各组VEGF、IL-1β、TNF-α的蛋白含量。结果 碱烧伤后第7、14天,低浓度石斛酚组、高浓度石斛酚组、阿柏西普组角膜新生血管面积占全角膜面积百分比均显著低于模型对照组(所有P<0.05)。碱烧伤后第14天,高浓度石斛酚组角膜混浊评分显著小于模型对照组(P<0.05),模型对照组、低浓度石斛酚组角膜厚度均显著高于正常对照组(所有P<0.001)。但是,高浓度石斛酚组、阿柏西普组角膜厚度与正常对照组相比较,均无显著性差异(所有P>0.05)。此外,低浓度石斛酚组、高浓度石斛酚组、阿柏西普组角膜组织中VEGF、IL-1β、TNF-α蛋白表达均显著低于模型对照组(所有P<0.01)。结论 结膜下注射石斛酚对碱烧伤大鼠角膜新生血管有抑制作用,并能促进角膜水肿的吸收。

塞来昔布固体脂质纳米粒对兔角膜重度热烧伤行穿透性角膜移植术后CNV、MMP-2的影响

目的观察塞来昔布(celecoxib,CXB)固体脂质纳米粒(solid lipid nanoparticles,SLN)对兔角膜重度热烧伤(severe corneal thermal burn,SCTB)行穿透性角膜移植术(penetrating keratoplasty,PKP)后角膜新生血管(corneal neovascularization,CNV)及基质金属蛋白酶2(matrix metalloproteinase-2,MMP-2)浓度的影响,探讨CXB-SLN对兔SCTB的作用机制。方法取36只新西兰白兔,恒温烧灼器制作兔SCTB模型(左眼),随机分为对照组、PKP组、CXB-PKP组,每组12只。对照组、PKP组球结膜下注射9 g/L的生理盐水0.1 ml,CXB-PKP组球结膜下注射1 g/L的CXB-SLN 0.1 ml。PKP组、CXB-PKP组于造模后第3天行PKP治疗。观察3组白兔术后角膜一般情况并记录术后10、20、40天CNV面积。在术后20、40天各组随机选择6只兔处死。采用酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)检测角膜上清液中MMP-2浓度,并分析CNV面积与MMP-2的相关性。结果术前3组白兔角膜均可见角膜缘充血明显,角膜烧伤区水肿,混浊,伴角膜上皮层坏死、剥脱。术后对照组可见角膜混浊逐渐加重呈瓷白色混浊,PKP组与CXB-PKP组可见角膜植片均成活,角膜透明度尚可。术后10、20、40天,CXB-PKP组CNV面积小于PKP组(P均<0.05),PKP组CNV面积小于对照组(P均<0.05)。术后20和40天角膜上清液MMP-2的浓度,CXB-PKP组低于PKP组,PKP组低于对照组(P均<0.05)。术后20、40天,CNV面积与MMP-2浓度呈正相关(r=0.742,P<0.001;r=0.827,P<0.001)。结论CXB-SLN对兔SCTB后的CNV具有明显抑制作用,可减轻角膜混浊,其机理可能与MMP-2的下调有关,能够为SCTB后行PKP创造有利条件。

诺帝滴眼液对碱烧伤诱导角膜新生血管的抑制作用

目的 观察自制诺帝滴眼液（Nordy eye drop）对Long．Evans大鼠角膜碱烧伤后新生血管（CRNV）有无抑制作用及其对正常角膜有无毒副作用。方法 采用0．025％、0．1％两个不同浓度诺帝滴眼液滴眼21d，通过裂隙灯、光镜、电镜观察检测诺帝滴眼液的毒副作用。建立角膜碱烧伤后角膜新生血管模型，0．025％、0．1％两个不同浓度诺帝滴眼液4次／d滴眼治疗，0．1％地塞米松磷酸钠滴眼液作为阳性对照、空白溶液作为阴性对照，观察诺帝治疗效果。结果 诺帝滴眼液滴眼21d后观察正常大鼠，无畏光、结膜充血等刺激症状，组织学检查显示结构正常，未见炎性细胞浸润。大鼠角膜碱烧伤后1d时仅见角膜缘血管网扩张，3d时新生血管长入，7～14d达高峰，14d时治疗组和地塞米松组新生血管开始退化；比较各组CRNV面积与角膜面积的比值，治疗组和阳性对照组均小于阴性对照组并有非常显著性差异（P〈0．01）。治疗组与阳性对照组间、0．025％与0．1％治疗组间均无显著性差异（P〉0．05）。结论 0．025％、0．1％的诺帝滴眼液局部滴眼无明显毒副作用，并且对CRNV有显著抑制作用。

角膜缘干细胞移植治疗兔眼表碱烧伤的实验研究

目的 :探讨眼表碱烧伤后行角膜缘干细胞移植术的疗效 ,并与单纯行角膜移植术组作比较。方法 :在 16只兔双眼上制作碱烧伤模型 ,1d后 ,右眼行异体板层角膜缘干细胞移植术 ,左眼行异体板层角膜移植术 ,术后观察 6月 ,根据术眼的角膜新生血管分级、植片的混浊度、水肿度来计算移植排斥反应指数 (RI) ,以RI值判断植片的存活情况。结果 :行异体板层角膜缘干细胞移植术的 16眼中有 14眼RI <9,治愈率达 87.5 % ;行单纯异体板层角膜移植的 16眼中 6眼RI值 <9,有效率 3 7.5 % ;两者相比有显著性差异 (P <0 .0 0 1)。两组的新生血管数、植片的混浊度、水肿度及RI值均有显著性差异。结论 :含干细胞的异体角膜缘移植治疗眼表碱烧伤的疗效明显优于单纯异体角膜移植 ,是一种较有效的方法。

重组人内皮抑素滴眼液抑制小鼠角膜新生血管形成的研究

目的 检验重组人内皮抑素对角膜新生血管形成的抑制效果,探索其临床应用价值。 方法 用MTT方法检验重组内皮抑素对血管内皮细胞的增殖抑制活性;用重组人内皮抑素滴眼液,治疗小鼠角膜碱烧伤导致的角膜新生血管形成。 结果 重组人内皮抑素 10μg/mL时可抑制bFGF刺激下HUVEC的增殖,抑制率达到 42 1% (P<0 05); 100μg/mL的内皮抑素可抑制碱烧伤诱导的小鼠角膜新生血管形成,治疗 7d时新生血管长度较未治疗组减少 30 9%(P<0 01),新生血管化面积减少了 13 7% (P<0 01)。 结论 内皮抑素特异地抑制血管内皮细胞增殖,抑制碱烧伤诱导的小鼠角膜新生血管形成,可能成为临床上预防、治疗相关眼科疾患的药物。

兔角膜碱烧伤后角膜基质注射脐带间充质干细胞的疗效

目的观察角膜基质注射脐带间充质干细胞(umbilicalcord mesenchymal stem cells,UMSCs)治疗兔角膜碱烧伤的效果。方法健康新西兰大白兔随机分为3组,每组12只,每只兔左眼为角膜碱烧伤模型眼。A组兔左眼在碱烧伤2 d后角膜基质内注射含有2×106个UMSCs的磷酸盐缓冲液2μL,B组注射相同剂量的磷酸盐缓冲液,C组为角膜碱烧伤后未进行处理组,在注射后不同时间对角膜混浊程度、新生血管生长情况、角膜上皮缺损情况等进行临床观察并评分,同时进行综合评分。结果注射后14 d A组新生血管生长较B组明显缓慢,A组、B组注射隧道周围角膜新生血管生长评分分别为0分、2分,差异有统计学意义(P<0.05)。注射后2 d和6 d A组角膜混浊程度较B组明显轻,注射后2 d A组、B组角膜混浊程度评分分别为2分、4分,注射后6 d评分与2 d相同,差异均具有统计学意义(均为P<0.05)。注射后2 d、7 d角膜上皮荧光素染色评分各组之间差异均无统学意义(均为P>0.05)。结论碱烧伤后角膜基质注射UMSCs可减少新生血管的形成,为抑制碱烧伤后角膜血管化提供参考,碱烧伤后角膜基质注射UMSCs可在一定程度上恢复角膜透明度。

电离辐射对角膜新生血管形成的抑制作用

背景角膜新生血管（CNV）可发生在多种眼表疾病中，常可加重病情，但有效的I临床治疗方法仍在探索中。目的探讨电离辐射对角膜碱烧伤后新生血管的抑制作用。方法76只清洁级Wistar纯系大鼠中70只用角膜碱烧伤的方法建立大鼠右眼CNV模型，然后按照随机数字表法将造模动物分为B射线10Gy一次性照射组2只、β射线7Gy分次照射组17只、β射线10Gy分次照射组17只、质量分数1％环孢素A（CsA）点眼组17只和角膜碱烧伤模型组17只，6只正常兔6只眼（均取右眼）作为正常对照组。用90Sr-90Y眼科敷贴器于右眼角膜碱烧伤后第1天开始沿角膜缘进行B射线照射，1％CsA点眼组用药时间与照射时间相同。实验后每日行裂隙灯检查并计算CNV长度和面积。于角膜碱烧伤后3、5、7d制备角膜组织石蜡切片和匀浆，采用免疫组织化学法检测各组大鼠角膜组织中bcl-2、bax、血管内皮生长因子（VEGF）的表达，分别采用Westernblot和逆转录聚合酶链反应（RT—PCR）检测各组大鼠角膜组织中VEGF蛋白及mRNA表达的变化。结果角膜碱烧伤后7d，裂隙灯下可见β射线10Gy一次性照射组、B射线10Gy分次照射组均出现角膜溃疡，角膜碱烧伤模型组可见大量CNV生成，而β射线7Gy分次照射组和1％CsA点眼组CNV明显较少。角膜碱烧伤后7d，β射线7cy分次照射组、1％CsA点眼组CNV长度和面积均明显小于角膜碱烧伤模型组，差异均有统计学意义（长度：q=14．40、24．20，P〈0．01；面积：q=17．80、14．00，P〈0．01）。免疫组织化学检测表明，与角膜碱烧伤模型组比较，β射线7Gy分次照射组、1％CsA点眼组大鼠角膜中bcl-2和VEGF蛋白表达均减弱，而bax蛋白表达均增强。RT—PCR检测表明，β射线7Gy分次照射组、β射线10Gy分次照射组、1％CsA点跟组VEGFmRNA表达强度明显低于角膜碱烧伤模型组，Westernblot检测发现VEGF蛋白的表达与VEGFmRNA的表达遵循同样的规律。结论90Sr-90Y眼科敷贴器小剂量分次放射治疗可明显抑制角膜碱烧伤后CNV的生长，并且以β射线7cy分次照射作用效果最佳。