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Impaired pericyte-Müller glia interaction via PDGFRβ suppression aggravates photoreceptor loss in a rodent model of light-induced retinal injury
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作者 Wei Xu Li-Jin Cui +3 位作者 Xiao-Ying Yang Xiao-Yuan Cui Jian Guo Guo-Xing Xu 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2024年第10期1800-1808,共9页
AIM:To investigate the involvement of pericyte-Müller glia interaction in retinal damage repair and assess the influence of suppressing the platelet-derived growth factor receptorβ(PDGFRβ)signaling pathway in r... AIM:To investigate the involvement of pericyte-Müller glia interaction in retinal damage repair and assess the influence of suppressing the platelet-derived growth factor receptorβ(PDGFRβ)signaling pathway in retinal pericytes on photoreceptor loss and Müller glial response.METHODS:Sprague-Dawley rats were exposed to intense light to induce retinal injury.Neutralizing antibody against PDGFRβwere deployed to block the signaling pathway in retinal pericytes through intravitreal injection.Retinal histology and Müller glial reaction were assessed following light injury.In vitro,normal and PDGFRβ-blocked retinal pericytes were cocultured with Müller cell line(rMC-1)to examine morphological and protein expression changes upon supplementation with light-injured supernatants of homogenized retinas(SHRs).RESULTS:PDGFRβblockage 24h prior to intense light exposure resulted in a significant exacerbation of photoreceptor loss.The upregulation of GFAP and p-STAT3,observed after intense light exposure,was significantly inhibited in the PDGFRβblockage group.Fur ther upregulation of cytokines monocyte chemoattractant protein 1(MCP-1)and interleukin-1β(IL-1β)was also observed following PDGFRβinhibition.In the in vitro coculture system,the addition of light-injured SHRs induced pericyte deformation and upregulation of proliferating cell nuclear antigen(PCNA)expression,while Müller cells exhibited neuron-like morphology and expressed Nestin.However,PDGFRβblockage in retinal pericytes abolished these cellular responses to light-induced damage,consistent with the in vivo PDGFRβblockage findings.CONCLUSION:Pericyte-Müller glia interaction plays a potential role in the endogenous repair process of retinal injury.Impairment of this interaction exacerbates photoreceptor degeneration in light-induced retinal injury. 展开更多
关键词 PERICYTE Müller glia light-induced retinal injury platelet-derived growth factor receptorβ signal pathway
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Effect of EGb761 on light-damaged retinal pigment epithelial cells
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作者 Yun-Yun Zhou Chang-Zheng Chen +5 位作者 Yu Su Lu Li Zuo-Hui-Zi Yi Hang Qi Ming Weng Yi-Qiao Xing 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2014年第1期8-13,共6页
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
关键词 EGB761 retinal pigment epithelial cells light damage PROTEIN mass spectrographic analysis
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Editor's Choice——Retinal ganglion cell damage and regeneration
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《Neural Regeneration Research》 SCIE CAS CSCD 2011年第31期2459-2459,共1页
Retinal ganglion cell apoptosis is considered to be the main cause of loss of vision in glaucoma patients. Microglia cells are phagocytic cells present in the retina. In the retina of glaucoma rat models, microglia ce... Retinal ganglion cell apoptosis is considered to be the main cause of loss of vision in glaucoma patients. Microglia cells are phagocytic cells present in the retina. In the retina of glaucoma rat models, microglia cells become activated, which suggests a role for microglia in the pathogenesis of optic nerve injury in glaucoma patients. The retinal ganglion cell is the only cell that can produce action potential in the retina, 展开更多
关键词 CELL Editor’s Choice retinal ganglion cell damage and regeneration
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Experimental model of photo-oxidative damage
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作者 Alessio Canovai 《Annals of Eye Science》 2022年第2期8-15,共8页
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
关键词 Photo-oxidative damage light-induced retinal damage photoreceptor death oxidative stress inflammation
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The Effect of Vaccinium Uliginosum on Rabbit Retinal Structure and Light-Induced Function Damage 被引量:3
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作者 尹澜 皮裕琍 张卯年 《Chinese Journal of Integrative Medicine》 SCIE CAS 2012年第4期299-303,共5页
Objective: To study the effect of Vaccinium uliginosum L., (VU) on the electroretinogram (ERG) and retinal pathological changes in rabbits after light-induced damage. Methods: Twenty-eight Chinchilla rabbits wer... Objective: To study the effect of Vaccinium uliginosum L., (VU) on the electroretinogram (ERG) and retinal pathological changes in rabbits after light-induced damage. Methods: Twenty-eight Chinchilla rabbits were randomly divided into four groups: administration beforehand (A), administration after injury (B), light injury without administration (C), and blank (D) groups. After a 4-week administration of VU homogenate at 4.8 g/(kg.d) once a day in group A, ERG in groups A, B and C were recorded according to the standards set by the International Society for Clinical Electrophysiology of Vision (ISCEV). Except for group D, the groups were then exposed to strong light. Just after that, group A stopped receiving VU treatment and group B started to receive it. Then ERGs in all groups were recorded after 1 day, 1 week, and 2 weeks. Throughout the whole process groups which were not fed with VU were fed with normal saline. Finally, the tissues and structures of all the groups were observed and the thickness of the outer nuclear layers (ONL) was measured. Results: (1) After 4-week feeding with VU, the latency time of ERG in group A became shorter than those in the other groups and the amplitude increased. After being exposed to strong light, the latency time lengthened and amplitude decreased in all the injury groups, but comparing at each time point, the measured values in group A were better than those in group C. With the accumulation of VU, the ERG in group B improved, and finally, all of the detected values became better than those in group C. (2) Retinae in group D were normal in histology and the layers were in order but those in group C became disarranged. The injudes in groups A and B were minor compared with those in group C. The thickness of the ONL in group C was significantly thinner than in the other groups (all P=0.000), and that in groups A and B was thicker than that in group C, although thinner than in group D. That in group A was thicker than in group B. Conclusions: VU can relieve the injury to rabbit retinae exposed to normal day and night rhythm, alleviate the harm caused by light when used beforehand, and repair the light damage to the retina. 展开更多
关键词 Vaccinium uliginosum L. light-induced damage ELECTROretinOGRAM HISTOLOGY
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Characteristics of Optic Nerve Damage Induced by Chronic Intraocular Hypertension in Rat 被引量:2
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作者 JiantaoWang JianGe +1 位作者 A.A.Sadun T.T.Lam 《Eye Science》 CAS 2004年第1期25-29,共5页
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
关键词 眼神经损害 慢性眼内高压 老鼠 RGCS 动物模型 视网膜神经节
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枸杞提取物对过氧化氢诱导的人视网膜色素上皮细胞氧化损伤的保护作用
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作者 黄冰林 魏含蕾 +4 位作者 黄琴 梁凌燕 唐秋雨 谢雨燕 袁灵梅 《江西中医药大学学报》 2024年第3期88-93,共6页
目的:体外通过过氧化氢(H2O2)诱导视网膜色素上皮(RPE)细胞造成氧化损伤,观察枸杞提取物对人视网膜色素上皮(ARPE-19)细胞活性的影响,探讨枸杞提取物通过抗氧化对ARPE-19细胞的保护作用。方法:采用不同浓度H2O2诱导体外ARPE-19细胞的氧... 目的:体外通过过氧化氢(H2O2)诱导视网膜色素上皮(RPE)细胞造成氧化损伤,观察枸杞提取物对人视网膜色素上皮(ARPE-19)细胞活性的影响,探讨枸杞提取物通过抗氧化对ARPE-19细胞的保护作用。方法:采用不同浓度H2O2诱导体外ARPE-19细胞的氧化损伤,确定最佳浓度;采用MTT法检测枸杞含药血清对ARPE-19细胞活力的影响,确定安全有效的药物浓度;观察各组细胞的形态,划痕试验检测各组ARPE-19细胞的活力;Western Blot法检测各组细胞Nrf2/ARE信号通路相关因子Nrf2的质蛋白和核蛋白的表达水平;试剂盒检测氧化应激水平的相关指标活性氧(ROS)含量、还原型谷胱甘肽(GSH)含量及过氧化氢酶(CAT)活力。结果:当H2O2浓度为200μmol/L,作用2 h时,ARPE-19细胞出现明显的氧化损伤(P<0.01);不同浓度的枸杞含药血清对ARPE-19细胞的干预效果不同,在2.5%、5%、10%的浓度下对损伤的ARPE-19细胞具有明显的促进增殖的作用(P<0.05),故选择2.5%、5%、10%浓度的枸杞含药血清进行下一步实验,与模型组相比,枸杞含药血清组细胞形态良好,长势均匀;细胞内的ROS含量显著降低(P<0.01),GSH含量明显上升(P<0.05),CAT活力显著升高(P<0.01);Nrf2质蛋白表达增强(P<0.05),核蛋白表达减弱(P<0.05)。结论:枸杞含药血清能够提高ARPE-19细胞自身的抗氧化能力,从而对ARPE-19细胞产生保护作用。 展开更多
关键词 年龄相关性黄斑变性 氧化损伤 视网膜色素上皮细胞 枸杞提取物 Nrf2/ARE信号通路
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视网膜血管几何参数与肾损害相关性的研究进展
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作者 刘芳 陈小鸟 +5 位作者 董哲毅 王倩 林雯文 黄惠 张睿敏 陈香美 《解放军医学院学报》 CAS 2024年第2期189-193,共5页
肾病严重影响患者生活质量,其确诊依赖于有创检查。视网膜血管和肾小球同属于微循环系统,视网膜血管参数的异常改变是微循环障碍的表现,可能提示肾损害。研究表明,视网膜血管直径、分形维数、弯曲度与提示肾损害的生物标志物有关,可成... 肾病严重影响患者生活质量,其确诊依赖于有创检查。视网膜血管和肾小球同属于微循环系统,视网膜血管参数的异常改变是微循环障碍的表现,可能提示肾损害。研究表明,视网膜血管直径、分形维数、弯曲度与提示肾损害的生物标志物有关,可成为预测和评估肾病进展的非侵入性指标,本文就视网膜血管几何参数与肾损害相关性的研究进行综述,为未来开发新的诊断工具和治疗方法提供依据。 展开更多
关键词 视网膜血管 几何参数 肾损害 慢性肾病 微循环
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蓝莓花青素调控Notch1/Hes-1通路对糖尿病大鼠视网膜神经节细胞氧化损伤的影响及机制研究
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作者 邢爽 赵伟 +2 位作者 刘丽 郭莹 郭硕 《生物医学工程与临床》 CAS 2024年第1期96-102,共7页
目的探讨蓝莓花青素(BA)对糖尿病大鼠视网膜神经节细胞(RGC)氧化损伤的影响及可能的作用机制。方法将50只雄性SD大鼠随机分为正常组(NC组,n=10)和造模组,造模组一次性腹腔注射链脲佐菌素(STZ)50 mg/kg,构建糖尿病模型,造模成功后再随机... 目的探讨蓝莓花青素(BA)对糖尿病大鼠视网膜神经节细胞(RGC)氧化损伤的影响及可能的作用机制。方法将50只雄性SD大鼠随机分为正常组(NC组,n=10)和造模组,造模组一次性腹腔注射链脲佐菌素(STZ)50 mg/kg,构建糖尿病模型,造模成功后再随机分为糖尿病模型组(DM组)、蓝莓花青素组(BA组)、DAPT组(Notch1通路抑制剂)和蓝莓花青素+DAPT组(BA+DAPT组),每组10只。BA组每天给予20 mg/kg的BA灌胃,DAPT组给予10μmol/L的DAPT干预,BA+DAPT组给予20 mg/kg的BA灌胃+10μmol/L的DAPT干预,NC组和DM组用等量0.9%氯化钠溶液(生理盐水)灌胃,连续8周。于末次给药24 h后,摘取大鼠眼球,用酶联免疫吸附分析(ELISA)试剂盒检测各组大鼠视网膜组织中活性氧(ROS)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)和丙二醛(MDA)含量;苏木精-伊红(HE)染色观察大鼠视网膜组织病理变化,并进行RGC计数;用荧光定量聚合酶链式反应(qRT-PCR)测量HO-1和iNOS mRNA表达;Western blot法检测Notch1和Hes-1蛋白的表达。结果建模后,DM组、BA组、DAPT组和BA+DAPT组空腹血糖(FBG)值均大于16.7 mmol/L;经BA干预治疗后,BA组和BA+DAPT组FBG含量明显降低,BA组低于BA+DAPT组(P<0.05)。ELISA结果表明,与NC组比较,DM组视网膜组织中ROS和MDA含量升高,CAT和SOD含量降低(P<0.05);与DM组比较,BA组和BA+DAPT组视网膜组织中ROS和MDA水平下降,CAT和SOD水平显著上升(P<0.05)。光学显微镜观察显示,NC组视网膜结构清晰,细胞形态正常,排列整齐;DM组大鼠RGC层排列紊乱,内、外核层伴随有水肿,出现了空泡现象,视网膜厚度变薄;BA组大鼠视网膜层结构排列较整齐,水肿程度和视网膜厚度较DM组有一定的改善;BA+DAPT组大鼠视网膜组织较DM组有所改善,但视网膜厚度相比BA组还是较薄。RGC计数结果显示,DM组、BA组、DAPT组和BA+DAPT组RGC数量分别为7.35±1.28、12.05±1.52、5.61±1.72、8.72±1.19,明显低于NC组的16.63±1.54,组间差异有统计学意义(F=88.905,P<0.05)。qRT-PCR结果表明,NC组、DM组、BA组、DAPT组和BA+DAPT组大鼠视网膜组织中HO-1水平分别为1.28±0.32、0.68±0.15、1.03±0.19、0.50±0.13和0.73±0.11,iNOS水平分别为0.48±0.08、1.17±0.21、0.89±0.17、1.35±0.15和1.10±0.23,差异均有统计学意义(F=25.192、35.843,P<0.05)。Western blot结果显示,与NC组相比,DM组视网膜组织中Notch1和Hes-1蛋白表达均下调;与DM组比较,BA组Notch1和Hes-1蛋白表达水平均上调,DAPT组Notch1和Hes-1蛋白表达水平均下调,BA+DAPT组两蛋白表达低于BA组,组间差异有统计学意义(F=47.626、54.709,P<0.05)。结论BA可以降低糖尿病大鼠FBG,增强其抗氧化应激能力,改善视网膜组织病理学损伤,从而对神经节细胞起到保护作用,其作用机制可能与激活Notch1/Hes-1信号通路有关。 展开更多
关键词 蓝莓花青素 糖尿病视网膜神经节细胞 氧化损伤 Notch1/Hes-1信号通路
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基于“伏毒损络”论治视网膜血管病
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作者 王丹 郝晓凤 +4 位作者 谢立科 孙梅 张孟姣 关小多 陈子扬 《世界中西医结合杂志》 2024年第8期1684-1688,共5页
视网膜血管病(Retinal vascular diseases,RVD)是包括糖尿病视网膜病变、视网膜静脉阻塞、视网膜动脉阻塞等在内的眼底血管性疾病的统称,常并发黄斑水肿和新生血管生成。RVD乃人体正气不足,脏腑功能失调,瘀、痰、湿、热等病理产物渐生,... 视网膜血管病(Retinal vascular diseases,RVD)是包括糖尿病视网膜病变、视网膜静脉阻塞、视网膜动脉阻塞等在内的眼底血管性疾病的统称,常并发黄斑水肿和新生血管生成。RVD乃人体正气不足,脏腑功能失调,瘀、痰、湿、热等病理产物渐生,随气血游弋至目络并深伏于此,日久蕴结化毒,毒损目络所致。RVD之起病隐匿、发病严重、病因复杂、容易反复、顽固难愈的临床特点与中医“伏毒”致病特点相一致,“正虚毒伏”为发病之始,“伏毒损络”乃发病关键,“伏毒留恋”致后期反复发作。因此,治疗RVD当以“扶正祛邪、解毒通络”为治疗总则,并在疾病发展的不同阶段予以不同的祛毒之法。此外,可适当佐以虫类药、藤类药等引药入络,直导毒巢。 展开更多
关键词 视网膜血管病 伏毒损络 分期论治 引药入络
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黄芪甲苷对人视网膜色素上皮细胞线粒体氧化损伤的调控作用
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作者 张艺曦 张茜癑 +1 位作者 白一辰 陆晓和 《世界中西医结合杂志》 2024年第10期1982-1986,共5页
目的探究黄芪甲苷对H_(2)O_(2)诱导人视网膜上皮细胞(Retinal pigment epithelium,RPE)线粒体氧化损伤的调控作用。方法建立H_(2)O_(2)诱导人RPEARPE-19和hTERT RPE-1氧化损伤模型,并加入不同剂量黄芪甲苷,用CCK-8法检测ARPE-19和hTERT ... 目的探究黄芪甲苷对H_(2)O_(2)诱导人视网膜上皮细胞(Retinal pigment epithelium,RPE)线粒体氧化损伤的调控作用。方法建立H_(2)O_(2)诱导人RPEARPE-19和hTERT RPE-1氧化损伤模型,并加入不同剂量黄芪甲苷,用CCK-8法检测ARPE-19和hTERT RPE-1细胞增殖活性,用荧光探针标记法检测ARPE-19细胞活性氧(Reactive oxygen,ROS)和脂质过氧化水平,用化学显色法检测ARPE-19细胞内SOD、LDH和MDA水平,用JC-1标记法检测ARPE-19细胞线粒体膜电势,用Calcein AM荧光染色法检测ARPE-19线粒体通透性,用蛋白质免疫印迹实验检测ARPE-19细胞Nrf2表达及磷酸化水平。结果与对照组比较,H_(2)O_(2)模型组ARPE-19和hTERT RPE-1细胞增殖活性明显下降(P<0.05),细胞内ROS积累增加(P<0.05),脂质过氧化水平升高(P<0.05),SOD和LDH表达水平下降(P<0.05),MDA水平升高(P<0.05),线粒体膜电势下降(P<0.05),通透性增加(P<0.05),Nrf2表达水平升高(P<0.05),磷酸化水平增加(P<0.05)。与H_(2)O_(2)模型组比较,低剂量、中剂量和高剂量黄芪甲苷处理的ARPE-19细胞增殖活性明显升高(P<0.05),中剂量和高剂量黄芪甲苷处理的hTERT RPE-1细胞增殖活性明显升高(P<0.05)。ARPE-19细胞内ROS和脂质过氧化水平下降(均P<0.05),SOD和LDH表达水平升高(P<0.05),MDA水平下降(P<0.05),线粒体膜电势升高(P<0.05),通透性降低(P<0.05),Nrf2表达无明显变化(P>0.05),磷酸化水平增加(P<0.05)。结论黄芪甲苷缓解H_(2)O_(2)诱导的人RPE氧化应激和线粒体功能损伤。 展开更多
关键词 黄芪甲苷 视网膜上皮细胞 氧化损伤 线粒体
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LED光源照射及PI3K抑制剂对人视网膜色素上皮细胞自噬和凋亡的影响
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作者 师若迪 徐晨 +1 位作者 李娟 俞洋 《浙江医学》 CAS 2024年第2期124-130,I0003,共8页
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
关键词 LED光源照射 PI3K抑制剂 视网膜色素上皮细胞 年龄相关性黄斑变性 光损伤 自噬 凋亡
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S100A8/S100A9在视网膜退行性疾病中的作用及机制研究进展
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作者 黄伟迪 陆才洋 +6 位作者 陈树明 唐子淳 李勰 郑淑燕 黄茜璇 刘骁 李卓 《国际眼科杂志》 CAS 2024年第10期1610-1614,共5页
S100蛋白家族属于损伤相关分子模式(DAMP),其在机体先天免疫反应中发挥着重要的炎症调节作用。其中S100A8/S100A9蛋白在众多疾病中发挥着广泛的抗菌、抗感染功能,并促进机体免疫及炎症反应的发生发展。在各类视网膜退行性疾病中,S100A8/... S100蛋白家族属于损伤相关分子模式(DAMP),其在机体先天免疫反应中发挥着重要的炎症调节作用。其中S100A8/S100A9蛋白在众多疾病中发挥着广泛的抗菌、抗感染功能,并促进机体免疫及炎症反应的发生发展。在各类视网膜退行性疾病中,S100A8/S100A9蛋白在转录及翻译阶段均明显上调,可促进眼部组织炎症因子的激活、巨噬细胞和中性粒细胞等免疫细胞的激活与募集,促进眼部炎症发生发展。文章旨在阐述S100A8/S100A9蛋白的生物学功能及其在视网膜退行性疾病如糖尿病视网膜病变、年龄相关性黄斑变性和缺血性视网膜病变中的作用及可能的机制。 展开更多
关键词 损伤相关分子模式(DAMP) S100A8/S100A9蛋白 糖尿病视网膜病变 炎症 视网膜退行性疾病
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抑制GSK-3β活性对糖尿病小鼠视网膜神经细胞损伤的影响
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作者 唐德荣 石蕊 +1 位作者 张阳炼 姜明玖 《临床眼科杂志》 2024年第4期350-354,共5页
目的探讨抑制糖原合成酶激酶-3β(GSK-3β)对糖尿病小鼠视网膜神经细胞损伤的影响,并探讨其潜在机制。方法前瞻性研究。健康雄性C57BL/6J小鼠,采用链脲佐菌素(STZ)腹腔注射构建糖尿病模型,视网膜HE染色及超微结构观察视网膜神经变性及... 目的探讨抑制糖原合成酶激酶-3β(GSK-3β)对糖尿病小鼠视网膜神经细胞损伤的影响,并探讨其潜在机制。方法前瞻性研究。健康雄性C57BL/6J小鼠,采用链脲佐菌素(STZ)腹腔注射构建糖尿病模型,视网膜HE染色及超微结构观察视网膜神经变性及神经细胞中线粒体损伤情况,Realtime PCR及免疫荧光检测视网膜中GSK-3βmRNA及蛋白的表达,并ELISA法检测视网膜中IL-6及IL-1β的水平变化。糖尿病小鼠玻璃体腔注射GSK-3β拮抗剂,观察其下游炎性因子的表达及视网膜神经元线粒体损伤情况。结果与糖尿病相比,GSK-3β阻断组糖尿病小鼠视网膜神经纤维层萎缩及神经元线粒体凋亡明显减轻,视网膜中炎性因子水平下降,组间比较差异有统计学意义(P<0.05)。结论阻断GSK-3β通过抑制炎症反应,减少神经元线粒体损伤,发挥糖尿病视网膜神经保护作用。 展开更多
关键词 糖尿病视网膜神经损伤 GSK-3Β 炎症 线粒体损伤
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糖尿病视网膜病变发病机制及潜在治疗靶点研究最新进展
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作者 刘秋平 刘昭麟 李晶明 《眼科新进展》 CAS 北大核心 2024年第10期757-768,共12页
糖尿病(DM)的患病率在全球范围内不断增加,DM的高发促使糖尿病视网膜病变(DR)患者数量激增,DR目前已成为工作年龄人群视力损害和失明的主要原因。DR的进行性发展,涉及许多分子、生物化学机制,而且各种机制之间相互作用,共同影响着视网... 糖尿病(DM)的患病率在全球范围内不断增加,DM的高发促使糖尿病视网膜病变(DR)患者数量激增,DR目前已成为工作年龄人群视力损害和失明的主要原因。DR的进行性发展,涉及许多分子、生物化学机制,而且各种机制之间相互作用,共同影响着视网膜血管及细胞的内环境稳态。随着对DR发病机制的深入研究,越来越多的分子靶点被发现。通过研究DR的发病机制,从中寻找潜在的治疗靶点,对于DR的早期预防、治疗以及防止疾病进一步进展,具有重大临床意义。本文对近年来DR的发病机制及潜在治疗靶点的研究进展进行简要综述。 展开更多
关键词 糖尿病视网膜病变 炎症 表观遗传修饰 线粒体损伤 视网膜神经变性 氧化应激及硝基化应激 微血管功能障碍 遗传多态性
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石斛碱对过氧化氢诱导的RGC-5细胞氧化损伤的保护作用
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作者 毛黔华 许凤青 +2 位作者 岳芳 张玮 石京山 《遵义医科大学学报》 2024年第8期768-774,782,共8页
目的研究石斛碱对过氧化氢(H_(2)O_(2))诱导的视神经节细胞氧化损伤的保护作用。方法将培养的RGC-5细胞分为正常对照组、H_(2)O_(2)模型组、石斛碱1 nmol/L组、石斛碱10 nmol/L组及石斛碱100 nmol/L组。采用细胞技术试剂盒-8(CCK-8)检测... 目的研究石斛碱对过氧化氢(H_(2)O_(2))诱导的视神经节细胞氧化损伤的保护作用。方法将培养的RGC-5细胞分为正常对照组、H_(2)O_(2)模型组、石斛碱1 nmol/L组、石斛碱10 nmol/L组及石斛碱100 nmol/L组。采用细胞技术试剂盒-8(CCK-8)检测RGC-5细胞活力;试剂盒法检测细胞中氧化和抗氧化物质的含量、细胞中ROS水平;蛋白免疫印迹法(Western Blot)检测细胞中核因子E2相关因子2(Nrf2)以及下游靶蛋白表达。结果石斛碱在1、10、100 nmol/L浓度对RGC-5细胞活力无毒性作用。H_(2)O_(2)从200μmol/L浓度开始显著降低RGC-5细胞活力。100 nmol/L石斛碱可显著降低H_(2)O_(2)诱导的RGC-5细胞中LDH外漏率、活性氧和丙二醛含量,显著升高超氧化物歧化酶和谷胱甘肽含量。石斛碱可诱导RGC-5细胞中Nrf2从胞浆向胞核转移,同时正向调控其下游抗氧化蛋白GPx1、GCLC、NQO 1和HO-1的蛋白表达。结论石斛碱对H_(2)O_(2)诱导的RGC-5细胞氧化损伤有明显的保护作用,其机制可能是石斛碱促进Nrf2核转位,正向调控Nrf2下游抗氧化蛋白表达发挥抗氧化作用。 展开更多
关键词 石斛碱 视神经节细胞 过氧化氢 氧化损伤
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Protective effects of a novel drug RC28-E blocking both VEGF and FGF2 on early diabetic rat retina 被引量:13
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作者 Qian-Hui Yang Yan Zhang +11 位作者 Jing Jiang Mian-Mian Wu Qian Han Qi-Yu Bo Guang-Wei Yu Yu-Sha Ru Xun Liu Min Huang Ling Wang Xiao-Min Zhang Jian-Min Fang Xiao-Rong Li 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2018年第6期935-944,共10页
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 ... 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
关键词 diabetic retinopathy vascular endothelialgrowth factor fibroblast growth factor 2 recombinant decoy receptor retinal damage diabetes
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Effects of perfluorooctane on the retina as a short-term and small amounts remnant in rabbits 被引量:1
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作者 Qing-Chen Li Jian Yu +3 位作者 Chun-Hui Jiang Hao-Hao Zhu Kun Liu Jin-Chuan Zhao 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2019年第3期381-386,共6页
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
关键词 perfluorooctane retinal damage TAMPONADE
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光诱导视网膜损伤的研究进展
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作者 刘佳 赵天祺 +1 位作者 牛金博 赵海霞 《内蒙古医科大学学报》 2023年第5期535-538,共4页
视网膜是眼组织中最容易受到光损伤的部位,过度暴露在强光下会造成不可逆的损伤。目前对光损伤视网膜的机制并没有一个明确的说明,可能是由于一系列的反应所堆积与微环境变化有关联。本文在视网膜内细胞层次,从视锥与视杆细胞、视网膜... 视网膜是眼组织中最容易受到光损伤的部位,过度暴露在强光下会造成不可逆的损伤。目前对光损伤视网膜的机制并没有一个明确的说明,可能是由于一系列的反应所堆积与微环境变化有关联。本文在视网膜内细胞层次,从视锥与视杆细胞、视网膜色素细胞以及神经节细胞三方面对光诱导视网膜的损伤进行综述。 展开更多
关键词 视网膜光损伤 光感受器 视网膜色素上皮细胞 视网膜神经节细胞
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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
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作者 李娟 陈雨婷 +2 位作者 师若迪 高园园 俞洋 《眼科新进展》 CAS 北大核心 2023年第8期594-598,共5页
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
关键词 视网膜色素上皮细胞 光损伤 细胞凋亡 氧化应激 miRNA-21-5p
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