Mitochondria-targeted antioxidant peptide SS31 attenuates high glucose-induced injury on human retinal endothelial cells
Mitochondria-targeted antioxidant peptide SS31 attenuates high glucose-induced injury on human retinal endothelial cells
复制标题
线粒体靶向抗氧化肽 SS31 减轻高糖诱导的人视网膜内皮细胞损伤
DOI：
10.1016/j.bbrc.2010.11.122
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发表时间：
2011-01-07
影响因子：
3.1
通讯作者：
Luo, Yan
中科院分区：
文献类型：
--
作者：
Li, Jianqiao;Chen, Xiaoyun;Luo, Yan
目的：探讨线粒体靶向抗氧化肽SS31对预防高糖诱导的人视网膜内皮细胞（HRECs）损伤的作用。方法：将培养的P3-P5 HRECs分为三组：5 mM葡萄糖组、30 mM葡萄糖组和30 mM葡萄糖与100 nM SS31联合处理组。治疗后24和48小时，使用Annexin V-FITC/PI染色评估HRECs的存活率。在共聚焦显微镜下通过 MitoSOX 染色评估 ROS 的过量产生。 JC-1荧光探针染色后，通过流式细胞术测量HRECs线粒体电位（Delta Psi（m））的变化。通过共聚焦显微镜和蛋白质印迹评估细胞色素c的释放。通过蛋白质印迹和实时PCR检测caspase-3和硫氧还蛋白-2（Trx-2）的表达。结果：与高糖组相比，100 nM 5531联合处理显着保护HRECs免受高糖诱导的损伤，减少线粒体中ROS的产生，稳定Delta Psi（m），减少细胞色素c从线粒体到细胞质的释放，降低caspase-3的表达并增加高糖处理的 HREC 中的 Trx-2。结论：SS31 通过稳定 Delta Psi(m)、减少 ROS 产生、阻止线粒体释放细胞色素 c、降低 caspase-3 的表达和增加 Trx-2 的表达，减轻高糖诱导的 HREC 损伤。我们的研究表明 SS31 可能成为糖尿病视网膜病变和其他氧化应激相关疾病的潜在新疗法。 (C) 2010 Elsevier Inc. 保留所有权利。
Purpose: To investigate the effect of mitochondria-targeted antioxidant peptide SS31 on prevention of high glucose-induced injury on human retinal endothelial cells (HRECs).Methods: Cultured P3-P5 HRECs were divided into three groups: 5 mM glucose group, 30 mM glucose group and 30 mM glucose co-treated with 100 nM SS31 group. 24 and 48 h after treatment, Annexin V-FITC/PI staining was used to evaluate the survival of HRECs. Overproduction of ROS was assessed by MitoSOX staining under confocal microscope. Change of mitochondrial potential (Delta Psi(m)) of HRECs was measured by flow cytometry after JC-1 fluorescent probe staining. Release of cytochrome c was assessed by confocal microscopy and western blot. Expression of caspase-3 and thioredoxin-2 (Trx-2) were measured by western blot and real-time PCR.Results: Compared to the high glucose group, co-treatment with 100 nM 5531 significantly protected HRECs from high glucose-induced injury, reduced the production of ROS in mitochondria, stabilized Delta Psi(m), decreased the release of cytochrome c from mitochondria to cytoplasm, decreased the expression of caspase-3 and increased the expression of Trx-2 in high glucose-treated HRECs.Conclusions: SS31 attenuates the high glucose-induced injuries on HRECs by stabilizing Delta Psi(m), decreasing ROS production, preventing the release of cytochrome c from mitochondria, decreasing the expression of caspase-3 and increasing the expression of Trx-2. Our study suggests that SS31 may be as a potential new treatment for diabetic retinopathy and other oxidative stress-related diseases. (C) 2010 Elsevier Inc. All rights reserved.