[1]白雪,罗艳,刘太祥,等.外源性硫化氢调节视神经萎缩相关蛋白1（OPA1）表达对视网膜缺血-再灌注损伤的保护作用[J].眼科新进展,2020,40(9):811-816.[doi:10.13389/j.cnki.rao.2020.0185]
　BAI Xue,LUO Yan,LIU Tai-Xiang,et al.Exogenous H2S protects against retinal ischemia-reperfusion injury by modulating optic atrophy 1(OPA1) expression[J].Recent Advances in Ophthalmology,2020,40(9):811-816.[doi:10.13389/j.cnki.rao.2020.0185]
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外源性硫化氢调节视神经萎缩相关蛋白1（OPA1）表达对视网膜缺血-再灌注损伤的保护作用/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
40卷
期数:
2020年9期
页码:
811-816
栏目:
实验研究
出版日期:
2020-09-05

文章信息/Info

Title:
Exogenous H2S protects against retinal ischemia-reperfusion injury by modulating optic atrophy 1(OPA1) expression
作者:
白雪; 罗艳; 刘太祥; 罗鑫
563003　贵州省遵义市， 遵义医科大学附属医院眼科
Author(s):
BAI Xue; LUO Yan; LIU Tai-Xiang; LUO Xin
Department of Ophthalmology,the Affiliated Hospital of Zunyi Medical University,Zunyi 563003,Guizhou Province,China
关键词:
硫化氢; OPA1; 视网膜缺血-再灌注损伤; 急性高眼压
Keywords:
hydrogen sulphide;  optic atrophy 1;  retinal ischemia-reperfusion injury;  acute high-eye pressure
分类号:
R775
DOI:
10.13389/j.cnki.rao.2020.0185
文献标志码:
A
摘要:
目的　探讨外源性硫化氢（H2S）对急性高眼压模型诱导视网膜缺血-再灌注损伤（retinal ischemia-reperfusion injury,RIRI)大鼠模型的作用和机制。方法　建立SD雄性大鼠急性高眼压模型（前房加压法）：通过自制的升眼压装置维持120 mmHg（1 kPa=7.5 mmHg）眼压，1 h后解除压力造成RIRI。硫氢化钠（NaHS）作为提供外源性H2S的供体，随机把52只健康无眼疾的SD雄性大鼠分为对照组（4只）、RIRI组（24只）及NaHS干预组（24只；连续5 d大鼠腹腔内注射NaHS液，造模前15 min再次给药）。后两组再分为造模后 1 h、6 h、12 h、24 h、48 h、72 h 6个时间点（n=4）。TUNEL法检测各组大鼠视网膜细胞凋亡指数，qPCR检测视网膜中视神经萎缩相关蛋白1（optic atrophy 1,OPA1）mRNA表达，Western blot检测各组大鼠视网膜细胞胞质以及线粒体中OPA1、CytC蛋白的表达，透射电镜观察各组线粒体形态。结果　与对照组比较：细胞凋亡指数除NaHS干预组造模后1 h（0.226±0.01）差异无统计学意义外（P＞0.05），RIRI组和NaHS干预组均增加（均为P<0.05），两组视网膜中OPA1 mRNA表达均下降（均为P<0.05），线粒体内OPA1和CytC的蛋白表达均下降（均为P<0.05），细胞质内均增加（均为P<0.05）。透射电镜下RIRI组和NaHS干预组线粒体肿胀，可见细胞质空泡及自噬小体。与RIRI组比较：NaHS干预组细胞凋亡减少（P<0.05）；视网膜OPA1 mRNA表达从造模后6 h开始均高于RIRI组（均为P<0.05），线粒体内OPA1和CytC表达从造模后24 h开始均高于RIRI组（均为P<0.05）；在细胞质内OPA1和CytC表达均低于RIRI组（均为P<0.05）；NaHS干预组各时间点线粒体损伤减轻。结论　H2S可能通过调节OPA1的表达与分布来减轻RIRI，且开始作用时间为造模后24 h。
Abstract:
Objective　To explore the role and possible mechanism of exogenous hydrogen sulfide (H2S) on rat models induced retinal ischemic re-perfusion damage (RIRI) in the acute high eye pressure model.Methods　An acute high eye pressure model of SD male rats was established by front room pressure method: 120 mmHg (1 kPa=7.5 mmHg) eye internal pressure maintained by a homemade eye pressure device, and the lifting of pressure after 1 hour causes RIRI. Sodium sulfide (NaHS) as a donor to provide exogenous H2S, 52 healthy, eye-free SD male rats were randomly divided into control group (4 rats), RIRI group (24 rats) and NaHS intervention group (24 rats) (the injection of NaHS solution into the intraperitoneal cavity of rats for 5 consecutive days, and re-administration 15 minutes before modeling). The latter two groups were then divided into six time periods of 1 h,6 h,12 h,24 h,48 h,72 h(4 rats for each time perior). TUNEL methods were used to detect the apoptotic index of rat retinal cells, qPCR to detect the expression of optic atrophy 1(OPA1) mRNA in the retina, Western blot to detect the expression of OPA1 protein and CytC in the cytoplasm and mitochondria of rat retinal cells in each group, and mitochondrial morphology was observed using transmission electron microscopy (TEM).Results　Compared with the control group, there was significantly more apoptosis was significantly increased in both experimental groups (all P< 0.05), except for the 1 h time pint group after NaHS intervention (0.226±0.010，P>0.05). OPA1 mRNA levels in the retinal cells decreased were significantly lower in both experimental groups (all P< 0.05), and OPA1 and CytC proteins expression were significantly decreased lower in the mitochondria (all P<0.05) and increased greater in the cytoplasm (all P< 0.05). Mitochondrial swelling, cytoplasmic vacuoles and autophagosomes were obvious under TEM. Comparison between groups: Apoptosis in the NaHS intervention group was reduced (P<0.05); the expression of OPA1mRNA in the retina was higher than that in the RIRI group 24 h after modeling (both P<0.05). The expressions of OPA1 and CytC proteins in the mitochondria was higher than those in the RIRI group 24 h after modeling (P<0.05). The expressions of OPA1 and CytC in the cytoplasm were all lower than those of the RRI group (both P＜0.05). Mitochondrial damage was significantly less at each time point in the NaHS intervention group.Conclusion　H2S may reduce the damage of acute high-eye pressure RIRI by regulating the expression and distribution of OPA1, and its effect occurs 24 hours after the modeling.

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备注/Memo

备注/Memo:
国家自然科学基金资助（编号：81860181）；贵州省科学技术基金资助(编号：20147588)
更新日期/Last Update: 2020-09-05