5,6-二氢环戊烯1，2-二硫杂环戊烯-3-硫酮对高糖环境下大鼠视网膜Muller细胞的影响  

Effect of 5,6-dihydrocyclopenta-1, 2-dithiole-3-thione on Müller cells under the high glucose

作　　者：黄棋 田敏 周琦 吕红彬 

机构地区：[1]西南医科大学附属医院眼科,泸州646000

出　　处：《中华眼底病杂志》2017年第3期290-294,共5页Chinese Journal of Ocular Fundus Diseases

摘　　要：目的 观察5,6-二氢环戊烯1,2-二硫杂环戊烯-3-硫酮（CPDT）对高糖环境下大鼠视网膜Müller细胞活性及其线粒体活性氧（ROS）生成量的影响，初步探讨CPDT对高糖环境下Müller细胞的保护作用及其机制。 方法 Sprague Dawley大鼠Müller细胞分为25 mmol/L对照组（A组）、65 mmol/L高糖（B糖）组、高糖＋45 μmol/L CPDT组（C组）、高糖＋60 μmol/L CPDT组（D组）、高糖＋70 μmol/L CPDT组（E组）。培养72 h后，水溶性四唑盐（WST）-8法检测各组Müller细胞的细胞相对增生率；流式细胞仪测定各组Müller细胞ROS生成量及细胞凋亡率；蛋白免疫印迹法（Western blot）测定Müller细胞中核因子-E2相关因子2（Nrf2）、血红素合酶-1（HO-1）、B细胞淋巴瘤/白血病-2（Bcl-2）和Bcl相关X蛋白（Bax蛋白）表达的变化。 结果 WST-8法检测结果显示，与A组比较，B组Müller细胞活性明显下降，差异有统计学意义（t=39.59，P＜0.05）。与B组比较，C组Müller细胞活性无明显提高，差异无统计学意义（t=0.97，P＞0.05）；D、E组Müller细胞活性恢复，差异有统计学意义（t=？4.67、？7.52，P＜0.05）。流式细胞仪检测结果显示，与A组比较，B组Müller细胞中ROS生成量增加，差异有统计学意义（t=–30.99，P＜0.05）；与B组比较，D组Müller细胞中ROS含量明显下降，差异有统计学意义（t=27.68，P＜0.05）。Western blot检测结果显示，与A组比较，B组Müller细胞Bax、Nrf2、HO-1蛋白表达显著上调，差异有统计学意义（t=？11.03、？63.17、？11.44，P＜0.05） ；Bcl-2蛋白表达明显下调，差异有统计学意义（t=7.861，P＜0.05）。与B组比较，D组Müller细胞Nrf2、HO-1、Bax蛋白表达均下调，差异有统计学意义（t=15.11、26.59、6.27，均P＜0.05）； Bcl-2蛋白表达上调，差异有统计学意义（t=？6.53，P＜0.05）。 结论 CPDT降低高糖环境下Müller细胞中ROS含量，下调Nrf2Objective To investigate the cellular viability and mitochondrial reactive oxygen species （ROS） production of the Müller cells under high glucose condition, and explore the protection role of the 5,6-dihydrocyclopenta-1, 2-dithiole-3-thione （CPDT） on Müller cells. Methods Müller cells from Sprague Dawley rats were divided into 5 groups randomly, including 25 mmol/L normal glucose group （group A） and 65 mmol/L high glucose group （group B）. High glucose group with 45, 60, 70 μmol/L CPDT and cultured them 72 hour was set as group C, D and E. Water soluble tetrazolium salt （WST）-8 was used to measure the cellular viability. Flow cytometry was used to measure the active oxygen and apoptosis index. The expression of nuclear factor erythroid 2-related factor 2 （Nrf2）, hemeoxygenase-1 （HO-1）, Bcl-2 and Bax protein were measured by Western blot. Results Compared with group A, the WST-8 showed that the viability of Müller cells apparently decreased in group B （t=39.59,P〈0.05）. Compared with the group B, the viability of Müller cells had changes in group C （t=0.97,P〉0.05）, but recovered in group D and E （t=？4.17, ？7.52;P〈0.05）. Compared with group A, the FCM showed that the mitochondrial ROS levels was higher in group B （t=？30.99,P〈0.05）. Compared with group B, the mitochondrial ROS levels were decreased in group D （t=27.68,P〈0.05）. Compared with group A, Bax, Nrf2 and HO-1 increased （t=–11.03, –63.17, –11.44;P〈0.05）, while the bcl-2 decreased in group B （t=7.861,P〈0.05）. Compared with the group B, Nrf2, HO-1 and Bax decreased （t=15.11, 26.59, 6.27;P〈0.05）, while the bcl-2 increased in group D （t=？6.53,P〈0.05）. Conclusions Under the high glucose, CPDT may reduce the mitochondrial ROS levels and the expression of Nrf2, HO-1 and Bax protein of Müller cells. It may inhibit apoptosis through activating the Nrf2/HO-1 pathway and balancing of level of Bcl-2 protein and mitochondrial ROS.

关 键 词：糖尿病视网膜病变/病理生理学 小神经胶质细胞 NF-E2相关因子2 血红素氧化酶 (脱环) BCL-2相关X蛋白质 

分 类 号：R587.2[医药卫生—内分泌] R774.1[医药卫生—内科学]