AIM: To investigate the protective mechanism of Gingko Biloba extract(EGb761) on the ability of retinal pigment epithelial(RPE) cells to resist light-induced damage in a comparative proteomics study. · METHODS: H...AIM: To investigate the protective mechanism of Gingko Biloba extract(EGb761) on the ability of retinal pigment epithelial(RPE) cells to resist light-induced damage in a comparative proteomics study. · METHODS: Human RPE cells(ARPE-19) were randomly distributed to one of three groups: normal control(NC group) and light-damaged model without or with EGb761 group(M and ME groups,respectively). The light-damaged model was formed by exposing to white light(2 200 ±300)lx for 6h. The RPE cells in ME group were conducted with EGb761(100μg/mL) before light exposure. The soluble cellular proteins extracting from each groups were separated by two-dimensional electrophoresis and stained by silver staining. Different proteins in the profiles of the gels were analyzed by Image Master Software. Two-fold expressing protein spots were identified by Matrix-assisted laser desorption/ ionization tandem time-of-flight(MALDI-TOF/TOF) mass spectrometry. ·RESULTS: NC,M and ME groups displayed 1 892±71,2 145 ±23 and 2 216 ±85 protein spots,respectively. We identified 33 proteins with different expression levels between the NC and M groups,25 proteins between the M and ME groups,and 11 proteins between the NC and ME groups. MALDI-TOF/TOF mass spectrometry successfully identified 16 proteins,including metabolic enzymes,cytoskeletal proteins,anti-oxidation proteins,and others. ·CONCLUSION: Differences in some important proteins,such as cathepsin B,heat shock protein,and cytochrome C reductase,indicated that multiple pathways may be induced in light-damaged RPE cells and the protective effect of EGb761.展开更多
Background:Retinal degeneration is a common feature of several retinal diseases,such as retinitis pigmentosa and age-related macular degeneration(AMD).In this respect,experimental models of photo-oxidative damage repr...Background:Retinal degeneration is a common feature of several retinal diseases,such as retinitis pigmentosa and age-related macular degeneration(AMD).In this respect,experimental models of photo-oxidative damage reproduce faithfully photoreceptor loss and many pathophysiological events involved in the activation of retinal cell degeneration.Therefore,such models represent a useful tool to study the mechanisms related to cell death.Their advantage consists in the possibility of modulating the severity of damage according to the needs of the experimenter.Indeed,bright light exposure could be regulated in both time and intensity to trigger a burst of apoptosis in photoreceptors,allowing the study of degenerative mechanisms in a controlled fashion,compared to the progressive and slower rate of death in other genetic models of photoreceptor degeneration.Methods:Here,an exemplificative protocol of bright light exposure in albino rat is described,as well as the main outcomes in retinal function,photoreceptor death,oxidative stress,and inflammation,which characterize this model and reproduce the main features of retinal degeneration diseases.Discussion:Models of photo-oxidative damage represent a useful tool to study the mechanisms responsible for photoreceptor degeneration.In this respect,it is important to adapt the exposure paradigm to the experimental needs,and the wide range of variables and limitations influencing the final outcomes should be considered to achieve proper results.Trial Registration:None.展开更多
Purpose:To set up the Sharma’s chronic intraocular hypertension model and investigate the intraocular pressure (IOP) as well as the optic nerve damage of this model in rat. Methods:The operations of the chronic intra...Purpose:To set up the Sharma’s chronic intraocular hypertension model and investigate the intraocular pressure (IOP) as well as the optic nerve damage of this model in rat. Methods:The operations of the chronic intraocular hypertension model were performed as described by Sharma in 60 male Lewis albino rats. IOP was measured using the Tono-Pen XL immediately after surgery and then at 5 day, 2 week or 4 week intervals. Cresyl violet staining of whole-mounted retinas was used to label retinal ganglion cells (RGCs), then RGCs were counted. Paraphenylenediamine (PPD) staining was performed in the semi-thin cross sections of optic nerve of rat, in order to know whether the axons of optic nerve were degenerated or not. Results:There were 47 rats with higher IOP after the episcleral veins cauterized in 60 rats. The ratio of elevated IOP was 78.3%. The IOPs were stable in 4 weeks. After cresyl violet staining, the RGCs loss was 11.0% and 11.3% was found in the central and peripheral retina respectively after 2 weeks of increased IOP. After 4 weeks of increased IOP, the loss of RGCs was 17% for the central retina and 24.6% for the peripheral retina. In the retinas without higher IOP, there was no loss of RGCs. PPD staining showed that optic nerve of rat with about 5.3% damage of axons located at the superior temporal region. Region of affected optic nerve 1 mm posterior to the globe by light microscope showed evidence of damaged axons with axonal swelling and myelin debris. Conclusion:Sharma’s chronic intraocular hypertension model is a reproducible and effective glaucoma model, which mimics human glaucoma with chronically elevation IOP and induced RGCs loss and damage of optic nerve. Eye Science 2004;20:25-29.展开更多
目的探讨LED光源照射对人视网膜色素上皮(RPE)细胞自噬和凋亡的影响,以及磷脂酰肌醇3激酶(PI3K)抑制剂LY294002在光照下对RPE细胞自噬和凋亡的影响。方法体外培养人ARPE-19细胞,随机分为6 h对照组、6 h模型组、6 h LY294002组,12 h对照...目的探讨LED光源照射对人视网膜色素上皮(RPE)细胞自噬和凋亡的影响,以及磷脂酰肌醇3激酶(PI3K)抑制剂LY294002在光照下对RPE细胞自噬和凋亡的影响。方法体外培养人ARPE-19细胞,随机分为6 h对照组、6 h模型组、6 h LY294002组,12 h对照组、12 h模型组、12 h LY294002组,24 h对照组、24 h模型组、24 h LY294002组,分别给于设定时间的LED冷光照射或加入LY294002后的光照。采用噻唑蓝法检测各组细胞存活率;流式细胞术检测各组细胞凋亡率;透射电子显微镜观察各组细胞超微结构;构建稳定表达红色荧光蛋白-绿色荧光蛋白-微管相关蛋白1轻链3(LC3)的RPE细胞株,激光共聚焦合倒置荧光显微镜分析细胞自噬流变化;Western blot法检测苄氯素1(Beclin 1)、LC3和p62自噬相关蛋白表达水平。结果与对照组比较,6、12、24 h模型组细胞存活率均下降(均P<0.05),12、24 h LY294002组细胞存活率均下降(均P<0.01);与模型组比较,12、24 h LY294002组细胞存活率均升高(均P<0.05)。与对照组比较,6、12、24 h模型组和LY294002组细胞凋亡率均升高(均P<0.05);与模型组比较,6、12 h LY294002组细胞凋亡率均下降(均P<0.05)。模型组RPE细胞在光照24 h后,胞内可见较多的自噬泡,LY294002干预后也可见聚集性分布的自噬囊泡。观察自噬流发现,对照组少见红色亮点,模型组红色亮点荧光随光照时间延长数量逐渐增多,Merge图中黄色亮点数量增多明显,LY294002干预后,红色亮点与黄色亮点呈增多趋势。与对照组比较,6、12、24 h模型组和LY294002组细胞中Beclin 1、LC3Ⅱ/LC3Ⅰ蛋白表达水平均升高,p62蛋白表达水平均下降,差异均有统计学意义(均P<0.05);与模型组比较,6、12、24 h LY294002组细胞中Beclin 1、LC3Ⅱ/LC3Ⅰ蛋白表达均升高,12、24 h LY294002组细胞中p62蛋白表达水平均下降,差异均有统计学意义(均P<0.05)。结论LED光源照射可刺激ARPE-19细胞发生自噬,增加细胞凋亡率;PI3K抑制剂LY294002能上调细胞的自噬活性,减缓凋亡。展开更多
AIM: To investigate protective effects of a novel recombinant decoy receptor drug RC28-E on retinal damage in early diabetic rats.METHODS: The streptozotocin(STZ)-induced diabetic rats were randomly divided into 6 gro...AIM: To investigate protective effects of a novel recombinant decoy receptor drug RC28-E on retinal damage in early diabetic rats.METHODS: The streptozotocin(STZ)-induced diabetic rats were randomly divided into 6 groups: diabetes mellitus(DM) group(saline, 3 μL/eye); RC28-E at low(0.33 μg/μL, 3 μL), medium(1 μg/μL, 3 μL), and high(3 μg/μL,3 μL) dose groups; vascular endothelial growth factor(VEGF) Trap group(1 μg/μL, 3 μL); fibroblast growth factor(FGF) Trap group(1 μg/μL, 3 μL). Normal control group was included. At week 1 and 4 following diabetic induction,the rats were intravitreally injected with the corresponding solutions. At week 6 following the induction, apoptosis in retinal vessels was detected by TUNEL staining.Glial fibrillary acidic protein(GFAP) expression was examined by immunofluorescence. Blood-retinal barrier(BRB) breakdown was assessed by Evans blue assay.Ultrastructural changes in choroidal and retinal vessels were analyzed by transmission electron microscopy(TEM).Content of VEGF and FGF proteins in retina was measured by enzyme linked immunosorbent assay(ELISA). The retinal expression of intercellular cell adhesion molecule-1(ICAM-1), tumor necrosis factor-α(TNF-α), VEGF and FGF genes was examined by quantitative polymerase chain reaction(qPCR).RESULTS: TUNEL staining showed that the aberrantly increased apoptotic cells death in diabetic retinal vascular network was significantly reduced by treatments of medium and high dose RC28-E, VEGF Trap, and FGF Trap(all P<0.05), the effects of medium and high dose RC28-E or FGF Trap were greater than VEGF Trap(P<0.01). GFAP staining suggested that reactive gliosis was substantially inhibited in all RC28-E and VEGF Trap groups, but the inhibition in FGF Trap group was not as prominent.Evans blue assay demonstrated that only high dose RC28-E could significantly reduce vascular leakage in early diabetic retina(P<0.01). TEM revealed that the ultrastructures in choroidal and retinal vessels were damaged in early diabetic retina, which was ameliorated to differential extents by each drug. The expression of VEGF and FGF2 proteins was significantly upregulated in early diabetic retina, and normalized by RC28-E at all dosages and by the corresponding Traps. The upregulation of ICAM-1 and TNF-α in diabetic retina was substantially suppressed by RC28-E and positive control drugs.CONCLUSION: Dual blockade of VEGF and FGF2 by RC28-E generates remarkable protective effects, including anti-apoptosis, anti-gliosis, anti-leakage, and improving ultrastructures and proinflammatory microenvironment,in early diabetic retina, thereby supporting further development of RC28-E into a novel and effective drug to diabetic retinopathy(DR).展开更多
AIM: To investigate changes in the rabbit retina after shortterm and small amounts tamponade of perfluorooctane(PFO).METHODS: New Zealand rabbits were used, and 48 eyes were randomly and evenly assigned into four diff...AIM: To investigate changes in the rabbit retina after shortterm and small amounts tamponade of perfluorooctane(PFO).METHODS: New Zealand rabbits were used, and 48 eyes were randomly and evenly assigned into four different groups. The PFO groups received a residue of 0.1 mL of PFO for ophthalmic surgery or 0.1 mL of F-Octane at the end of surgery; eyes from the pars plana vitrectomy(PPV) group were filled with balanced salt solution and those having not received surgical intervention served as controls. Eyes were collected at 1, 4 and 12 wk and studied.RESULTS: Under a microscope, nuclear counts of the inner nuclear layer(INL) and outer nuclear layer(ONL) did not differ among the four groups at all time points; however, slight disarrangement of the ONL and occasional vacuolization of the INL were found in the inferior retina only at 12 wk in two PFO groups. Four of the groups had similar results of Caspase-3 and TNF-α staining at all time points. Alternatively, IL-8 was increased in PFOa and PPV control groups at 4 wk and in all three PPV groups at 12 wk; also, the apoptotic index(%) was similarly increased in all three PPV groups at 4 and 12 wk.CONCLUSION: Both PFOs are well tolerated in rabbit eyes for up to 12 wk, which suggests that they can be used safely as intraoperative tools or for short-term and small amounts tamponade after surgery.展开更多
目的研究miRNA-21-5p对光诱导的人视网膜色素上皮细胞氧化应激损伤的影响。方法将体外培养的人视网膜色素上皮细胞系ARPE-19细胞随机分为对照组(TransIntro EL Transfection Reagent转染液培养)、损伤组(TransIntro EL Transfection Rea...目的研究miRNA-21-5p对光诱导的人视网膜色素上皮细胞氧化应激损伤的影响。方法将体外培养的人视网膜色素上皮细胞系ARPE-19细胞随机分为对照组(TransIntro EL Transfection Reagent转染液培养)、损伤组(TransIntro EL Transfection Reagent转染液+光损伤)、过表达组(TransIntro EL Transfection Reagent转染液+miRNA-21-5p mimics+光损伤)、阴性组(TransIntro EL Transfection Reagent转染液+miRNA-21-5p mimics NC+光损伤)、PI3K/Akt阻断剂组(TransIntro EL Transfection Reagent转染液+miRNA-21-5p mimics+LY294002+光损伤)。使用光照强度为(16500±200)lx的LED冷光灯建立ARPE-19细胞光损伤模型,利用TransIntro EL Transfection Reagent转染液行细胞转染。采用qRT-PCR法检测各组ARPE-19细胞miRNA-21-5p表达水平,采用CCK-8法检测各组ARPE-19细胞活力,流式细胞仪检测各组ARPE-19细胞活性氧(ROS)含量变化,ELISA法检测各组ARPE-19细胞超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量。结果与对照组相比,损伤组ARPE-19细胞miRNA-21-5p表达明显下降,细胞存活率明显下降,ROS含量显著升高,SOD活性明显降低,MDA含量明显增加(均为P<0.001);与损伤组相比,过表达组ARPE-19细胞miRNA-21-5p表达明显升高,细胞存活率明显上升,ROS含量明显降低,SOD活性升高,MDA含量减少(均为P<0.001),而阴性组ARPE-19细胞miRNA-21-5p表达、细胞存活率、ROS含量、SOD活性、MDA含量均无明显差异(均为P>0.05);与过表达组相比,PI3K/Akt阻断剂组ARPE-19细胞miRNA-21-5p表达明显降低,细胞存活率明显下降,ROS含量明显升高,SOD活性明显降低,MDA含量明显增加(均为P<0.01)。结论miRNA-21-5p能显著降低光诱导的ARPE-19细胞氧化应激水平,提高光诱导的ARPE-19细胞抗氧化能力。展开更多
基金Supported by National Natural Science Foundation of China(No.30500676)
文摘AIM: To investigate the protective mechanism of Gingko Biloba extract(EGb761) on the ability of retinal pigment epithelial(RPE) cells to resist light-induced damage in a comparative proteomics study. · METHODS: Human RPE cells(ARPE-19) were randomly distributed to one of three groups: normal control(NC group) and light-damaged model without or with EGb761 group(M and ME groups,respectively). The light-damaged model was formed by exposing to white light(2 200 ±300)lx for 6h. The RPE cells in ME group were conducted with EGb761(100μg/mL) before light exposure. The soluble cellular proteins extracting from each groups were separated by two-dimensional electrophoresis and stained by silver staining. Different proteins in the profiles of the gels were analyzed by Image Master Software. Two-fold expressing protein spots were identified by Matrix-assisted laser desorption/ ionization tandem time-of-flight(MALDI-TOF/TOF) mass spectrometry. ·RESULTS: NC,M and ME groups displayed 1 892±71,2 145 ±23 and 2 216 ±85 protein spots,respectively. We identified 33 proteins with different expression levels between the NC and M groups,25 proteins between the M and ME groups,and 11 proteins between the NC and ME groups. MALDI-TOF/TOF mass spectrometry successfully identified 16 proteins,including metabolic enzymes,cytoskeletal proteins,anti-oxidation proteins,and others. ·CONCLUSION: Differences in some important proteins,such as cathepsin B,heat shock protein,and cytochrome C reductase,indicated that multiple pathways may be induced in light-damaged RPE cells and the protective effect of EGb761.
文摘Background:Retinal degeneration is a common feature of several retinal diseases,such as retinitis pigmentosa and age-related macular degeneration(AMD).In this respect,experimental models of photo-oxidative damage reproduce faithfully photoreceptor loss and many pathophysiological events involved in the activation of retinal cell degeneration.Therefore,such models represent a useful tool to study the mechanisms related to cell death.Their advantage consists in the possibility of modulating the severity of damage according to the needs of the experimenter.Indeed,bright light exposure could be regulated in both time and intensity to trigger a burst of apoptosis in photoreceptors,allowing the study of degenerative mechanisms in a controlled fashion,compared to the progressive and slower rate of death in other genetic models of photoreceptor degeneration.Methods:Here,an exemplificative protocol of bright light exposure in albino rat is described,as well as the main outcomes in retinal function,photoreceptor death,oxidative stress,and inflammation,which characterize this model and reproduce the main features of retinal degeneration diseases.Discussion:Models of photo-oxidative damage represent a useful tool to study the mechanisms responsible for photoreceptor degeneration.In this respect,it is important to adapt the exposure paradigm to the experimental needs,and the wide range of variables and limitations influencing the final outcomes should be considered to achieve proper results.Trial Registration:None.
文摘Purpose:To set up the Sharma’s chronic intraocular hypertension model and investigate the intraocular pressure (IOP) as well as the optic nerve damage of this model in rat. Methods:The operations of the chronic intraocular hypertension model were performed as described by Sharma in 60 male Lewis albino rats. IOP was measured using the Tono-Pen XL immediately after surgery and then at 5 day, 2 week or 4 week intervals. Cresyl violet staining of whole-mounted retinas was used to label retinal ganglion cells (RGCs), then RGCs were counted. Paraphenylenediamine (PPD) staining was performed in the semi-thin cross sections of optic nerve of rat, in order to know whether the axons of optic nerve were degenerated or not. Results:There were 47 rats with higher IOP after the episcleral veins cauterized in 60 rats. The ratio of elevated IOP was 78.3%. The IOPs were stable in 4 weeks. After cresyl violet staining, the RGCs loss was 11.0% and 11.3% was found in the central and peripheral retina respectively after 2 weeks of increased IOP. After 4 weeks of increased IOP, the loss of RGCs was 17% for the central retina and 24.6% for the peripheral retina. In the retinas without higher IOP, there was no loss of RGCs. PPD staining showed that optic nerve of rat with about 5.3% damage of axons located at the superior temporal region. Region of affected optic nerve 1 mm posterior to the globe by light microscope showed evidence of damaged axons with axonal swelling and myelin debris. Conclusion:Sharma’s chronic intraocular hypertension model is a reproducible and effective glaucoma model, which mimics human glaucoma with chronically elevation IOP and induced RGCs loss and damage of optic nerve. Eye Science 2004;20:25-29.
文摘目的探讨LED光源照射对人视网膜色素上皮(RPE)细胞自噬和凋亡的影响,以及磷脂酰肌醇3激酶(PI3K)抑制剂LY294002在光照下对RPE细胞自噬和凋亡的影响。方法体外培养人ARPE-19细胞,随机分为6 h对照组、6 h模型组、6 h LY294002组,12 h对照组、12 h模型组、12 h LY294002组,24 h对照组、24 h模型组、24 h LY294002组,分别给于设定时间的LED冷光照射或加入LY294002后的光照。采用噻唑蓝法检测各组细胞存活率;流式细胞术检测各组细胞凋亡率;透射电子显微镜观察各组细胞超微结构;构建稳定表达红色荧光蛋白-绿色荧光蛋白-微管相关蛋白1轻链3(LC3)的RPE细胞株,激光共聚焦合倒置荧光显微镜分析细胞自噬流变化;Western blot法检测苄氯素1(Beclin 1)、LC3和p62自噬相关蛋白表达水平。结果与对照组比较,6、12、24 h模型组细胞存活率均下降(均P<0.05),12、24 h LY294002组细胞存活率均下降(均P<0.01);与模型组比较,12、24 h LY294002组细胞存活率均升高(均P<0.05)。与对照组比较,6、12、24 h模型组和LY294002组细胞凋亡率均升高(均P<0.05);与模型组比较,6、12 h LY294002组细胞凋亡率均下降(均P<0.05)。模型组RPE细胞在光照24 h后,胞内可见较多的自噬泡,LY294002干预后也可见聚集性分布的自噬囊泡。观察自噬流发现,对照组少见红色亮点,模型组红色亮点荧光随光照时间延长数量逐渐增多,Merge图中黄色亮点数量增多明显,LY294002干预后,红色亮点与黄色亮点呈增多趋势。与对照组比较,6、12、24 h模型组和LY294002组细胞中Beclin 1、LC3Ⅱ/LC3Ⅰ蛋白表达水平均升高,p62蛋白表达水平均下降,差异均有统计学意义(均P<0.05);与模型组比较,6、12、24 h LY294002组细胞中Beclin 1、LC3Ⅱ/LC3Ⅰ蛋白表达均升高,12、24 h LY294002组细胞中p62蛋白表达水平均下降,差异均有统计学意义(均P<0.05)。结论LED光源照射可刺激ARPE-19细胞发生自噬,增加细胞凋亡率;PI3K抑制剂LY294002能上调细胞的自噬活性,减缓凋亡。
文摘AIM: To investigate protective effects of a novel recombinant decoy receptor drug RC28-E on retinal damage in early diabetic rats.METHODS: The streptozotocin(STZ)-induced diabetic rats were randomly divided into 6 groups: diabetes mellitus(DM) group(saline, 3 μL/eye); RC28-E at low(0.33 μg/μL, 3 μL), medium(1 μg/μL, 3 μL), and high(3 μg/μL,3 μL) dose groups; vascular endothelial growth factor(VEGF) Trap group(1 μg/μL, 3 μL); fibroblast growth factor(FGF) Trap group(1 μg/μL, 3 μL). Normal control group was included. At week 1 and 4 following diabetic induction,the rats were intravitreally injected with the corresponding solutions. At week 6 following the induction, apoptosis in retinal vessels was detected by TUNEL staining.Glial fibrillary acidic protein(GFAP) expression was examined by immunofluorescence. Blood-retinal barrier(BRB) breakdown was assessed by Evans blue assay.Ultrastructural changes in choroidal and retinal vessels were analyzed by transmission electron microscopy(TEM).Content of VEGF and FGF proteins in retina was measured by enzyme linked immunosorbent assay(ELISA). The retinal expression of intercellular cell adhesion molecule-1(ICAM-1), tumor necrosis factor-α(TNF-α), VEGF and FGF genes was examined by quantitative polymerase chain reaction(qPCR).RESULTS: TUNEL staining showed that the aberrantly increased apoptotic cells death in diabetic retinal vascular network was significantly reduced by treatments of medium and high dose RC28-E, VEGF Trap, and FGF Trap(all P<0.05), the effects of medium and high dose RC28-E or FGF Trap were greater than VEGF Trap(P<0.01). GFAP staining suggested that reactive gliosis was substantially inhibited in all RC28-E and VEGF Trap groups, but the inhibition in FGF Trap group was not as prominent.Evans blue assay demonstrated that only high dose RC28-E could significantly reduce vascular leakage in early diabetic retina(P<0.01). TEM revealed that the ultrastructures in choroidal and retinal vessels were damaged in early diabetic retina, which was ameliorated to differential extents by each drug. The expression of VEGF and FGF2 proteins was significantly upregulated in early diabetic retina, and normalized by RC28-E at all dosages and by the corresponding Traps. The upregulation of ICAM-1 and TNF-α in diabetic retina was substantially suppressed by RC28-E and positive control drugs.CONCLUSION: Dual blockade of VEGF and FGF2 by RC28-E generates remarkable protective effects, including anti-apoptosis, anti-gliosis, anti-leakage, and improving ultrastructures and proinflammatory microenvironment,in early diabetic retina, thereby supporting further development of RC28-E into a novel and effective drug to diabetic retinopathy(DR).
基金Supported by the National Key Research&Development Plan(No.2017YFC0108200)the Shanghai Committee of Science and Technology(No.16140901000+1 种基金 No.13430710500 No.15DZ1942204)
文摘AIM: To investigate changes in the rabbit retina after shortterm and small amounts tamponade of perfluorooctane(PFO).METHODS: New Zealand rabbits were used, and 48 eyes were randomly and evenly assigned into four different groups. The PFO groups received a residue of 0.1 mL of PFO for ophthalmic surgery or 0.1 mL of F-Octane at the end of surgery; eyes from the pars plana vitrectomy(PPV) group were filled with balanced salt solution and those having not received surgical intervention served as controls. Eyes were collected at 1, 4 and 12 wk and studied.RESULTS: Under a microscope, nuclear counts of the inner nuclear layer(INL) and outer nuclear layer(ONL) did not differ among the four groups at all time points; however, slight disarrangement of the ONL and occasional vacuolization of the INL were found in the inferior retina only at 12 wk in two PFO groups. Four of the groups had similar results of Caspase-3 and TNF-α staining at all time points. Alternatively, IL-8 was increased in PFOa and PPV control groups at 4 wk and in all three PPV groups at 12 wk; also, the apoptotic index(%) was similarly increased in all three PPV groups at 4 and 12 wk.CONCLUSION: Both PFOs are well tolerated in rabbit eyes for up to 12 wk, which suggests that they can be used safely as intraoperative tools or for short-term and small amounts tamponade after surgery.
文摘目的研究miRNA-21-5p对光诱导的人视网膜色素上皮细胞氧化应激损伤的影响。方法将体外培养的人视网膜色素上皮细胞系ARPE-19细胞随机分为对照组(TransIntro EL Transfection Reagent转染液培养)、损伤组(TransIntro EL Transfection Reagent转染液+光损伤)、过表达组(TransIntro EL Transfection Reagent转染液+miRNA-21-5p mimics+光损伤)、阴性组(TransIntro EL Transfection Reagent转染液+miRNA-21-5p mimics NC+光损伤)、PI3K/Akt阻断剂组(TransIntro EL Transfection Reagent转染液+miRNA-21-5p mimics+LY294002+光损伤)。使用光照强度为(16500±200)lx的LED冷光灯建立ARPE-19细胞光损伤模型,利用TransIntro EL Transfection Reagent转染液行细胞转染。采用qRT-PCR法检测各组ARPE-19细胞miRNA-21-5p表达水平,采用CCK-8法检测各组ARPE-19细胞活力,流式细胞仪检测各组ARPE-19细胞活性氧(ROS)含量变化,ELISA法检测各组ARPE-19细胞超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量。结果与对照组相比,损伤组ARPE-19细胞miRNA-21-5p表达明显下降,细胞存活率明显下降,ROS含量显著升高,SOD活性明显降低,MDA含量明显增加(均为P<0.001);与损伤组相比,过表达组ARPE-19细胞miRNA-21-5p表达明显升高,细胞存活率明显上升,ROS含量明显降低,SOD活性升高,MDA含量减少(均为P<0.001),而阴性组ARPE-19细胞miRNA-21-5p表达、细胞存活率、ROS含量、SOD活性、MDA含量均无明显差异(均为P>0.05);与过表达组相比,PI3K/Akt阻断剂组ARPE-19细胞miRNA-21-5p表达明显降低,细胞存活率明显下降,ROS含量明显升高,SOD活性明显降低,MDA含量明显增加(均为P<0.01)。结论miRNA-21-5p能显著降低光诱导的ARPE-19细胞氧化应激水平,提高光诱导的ARPE-19细胞抗氧化能力。
