[1]边思林,刘华.α-硫辛酸对青光眼模型大鼠视网膜神经节细胞的保护作用及其机制[J].眼科新进展,2020,40(4):307-312.[doi:10.13389/j.cnki.rao.2020.0071]
　BIAN Silin,LIU Hua.Protective effect of α-lipoic acid on retinal ganglion cells in glaucoma model rats and its mechanisms[J].Recent Advances in Ophthalmology,2020,40(4):307-312.[doi:10.13389/j.cnki.rao.2020.0071]
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α-硫辛酸对青光眼模型大鼠视网膜神经节细胞的保护作用及其机制/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
40卷
期数:
2020年4期
页码:
307-312
栏目:
实验研究
出版日期:
2020-04-05

文章信息/Info

Title:
Protective effect of α-lipoic acid on retinal ganglion cells in glaucoma model rats and its mechanisms
作者:
边思林; 刘华
121001　辽宁省锦州市，锦州医科大学附属第三医院眼科
Author(s):
BIAN Silin; LIU Hua
Department of Ophthalmology,the Third Affiliated Hospital,Jinzhou Medical University,Jinzhou 121001,Liaoning Province,China
关键词:
α-硫辛酸; 青光眼; 视网膜神经节细胞; 氧化应激; 凋亡
Keywords:
α-lipoic acid;  glaucoma;  retinal ganglion cells;  oxidative stress;  apoptosis
分类号:
R775
DOI:
10.13389/j.cnki.rao.2020.0071
文献标志码:
A
摘要:
目的　探讨α-硫辛酸对青光眼模型大鼠视网膜神经节细胞（RGC）的保护作用及其机制。方法　选取SPF 级雄性SD大鼠 60只，取30只大鼠，选右眼为实验眼，采用烙闭表层巩膜静脉的方法制作大鼠青光眼模型。将造模成功的大鼠随机分为两组：青光眼对照组和青光眼硫辛酸组，每组各15眼。另取30只大鼠，右眼行假手术操作，术后随机分为两组：正常眼压对照组和正常眼压硫辛酸组，每组各15眼。于造模第3周末，正常眼压硫辛酸组和青光眼硫辛酸组大鼠腹腔注射α-硫辛酸150 mg·kg-1。分别于造模前、造模后第1周末、第2周末、第3周末、第4周末、第5周末、第6周末应用手持式眼压计测量并记录大鼠双眼眼压。分别用相应测试盒检测丙二醛(MDA)和活性氧簇(ROS)含量，超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶（GSH-Px）活性。通过TUNEL法检测RGC凋亡情况，通过免疫组织化学法检测视网膜Caspase-3 蛋白表达情况。应用GraphPadPrism6软件作图，采用SPSS 20.0统计软件分析所有数据。结果　造模后3周末开始腹腔注射α-硫辛酸，青光眼硫辛酸组大鼠右眼眼压在造模后4周、5周、6周与青光眼对照组比较，差异均无统计学意义（均为P＞0.05）。MDA和ROS含量：四组间比较差异均有统计学意义（均为P＜0.05），均为正常眼压硫辛酸组＜正常眼压对照组＜青光眼硫辛酸组＜青光眼对照组（均为P＜0.05）。GSH-Px及SOD酶活性：四组间比较差异均有统计学意义（均为P＜0.05），均为正常眼压硫辛酸组＞正常眼压对照组＞青光眼硫辛酸组＞青光眼对照组（均为P＜0.05）。青光眼硫辛酸组和青光眼对照组的凋亡细胞数明显高于正常眼压硫辛酸组和正常眼压对照组。正常眼压硫辛酸组RGC 凋亡指数和正常眼压对照组差异无统计学意义（P>0.05），但均小于青光眼硫辛酸组和青光眼对照组（均为P＜0.05）；青光眼硫辛酸组RGC 凋亡指数小于青光眼对照组（P＜0.05）。青光眼硫辛酸组和青光眼对照组Caspase-3蛋白表达明显高于正常眼压硫辛酸组和正常眼压对照组。青光眼硫辛酸组Caspase-3蛋白平均光密度值小于青光眼对照组（P＜0.05）。结论　α-硫辛酸通过抗氧化作用降低Caspase-3的表达，抑制RGC凋亡，对青光眼模型大鼠RGC具有一定的保护作用。
Abstract:
Objective　To investigate the protective effect of α-lipoic acid on retinal ganglion cells (RGC) in glaucoma model rats and its mechanisms.Methods　Sixty SPF male SD rats were selected. Right eyes of 30 rats were selected as the experimental group, and used to make glaucoma models by sealing the superficial scleral vein. The successful glaucoma rat models were randomly divided into two groups: glaucoma control group and glaucoma lipoic acid group, 15 eyes in each group. Right eyes of another 30 rats received sham operation, and were randomly divided into two groups after operation: normal intraocular pressure (IOP) control group and normal IOP lipoic acid group, 15 eyes in each group. At the end of the third week after modeling, rats in normal IOP lipoic acid group and glaucoma lipoic acid group were intraperitoneally injected with 150 mg·kg-1 of α-lipoic acid. The handheld tonometer was used to measure and record IOP of rats before modeling, at the end of the first, second, third, fourth, fifth and sixth weeks. The contents of malondialdehyde (MDA) and reactive oxygen species (ROS), and activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were measured by corresponding test boxes. The apoptosis of RGC was detected by TUNEL method, and the expression of Caspase-3 protein was detected by immunohistochemistry. GraphPadPrism6 software was used for mapping, and all data was analyzed using SPSS 20.0 statistical software.Results　Intraperitoneal injection of α-lipoic acid was started at the end of the third week after modeling, and IOP of the right eyes of rats in glaucoma lipoic acid group were not significantly different from those of rats in glaucoma control group at 4 weeks, 5 weeks and 6 weeks after modeling (all P>0.05). Levels of MDA and ROS: Statistical differences were observed for pairwise comparison (all P<0.05), and normal IOP lipoic acid group normal IOP control group > glaucoma lipoic acid group > glaucoma control group (all P<0.05). The number of apoptotic cells in the glaucoma lipoic acid group and the glaucoma control group were significantly higher than those in the normal IOP lipoic acid group and the normal IOP control group. There was no significant difference in the RGC apoptosis index between normal IOP lipoic acid group and normal IOP control group (P> 0.05), but both were lower than those in glaucoma lipoic acid group and glaucoma control group (all P<0.05). The apoptosis index of RGC in the glaucoma lipoic acid group was lower than that in the glaucoma control group (P<0.05). The expression of Caspase-3 protein in the glaucoma lipoic acid group and glaucoma control group were significantly higher than those in normal IOP lipoic acid group and normal IOP control group. The average optical density of Caspase-3 protein in the glaucoma lipoic acid group was lower than that in the glaucoma control group (P<0.05).Conclusion　α-lipoic acid reduces the expression of Caspase-3 and inhibits RGC apoptosis through anti-oxidation, which has a certain protective effect on RGC in glaucoma model rats.

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备注/Memo

备注/Memo:
国家自然科学基金资助(编号：31571184)；辽宁省科技厅锦州医科大学联合基金项目(编号：20170540382)
更新日期/Last Update: 2020-04-05