二项酶诱导剂对高糖环境下大鼠Müller细胞凋亡的影响  

Effects of a two-enzyme inducer on apoptosis in rat Müller cells in the high glucose environment

作　　者：吴晓强[1] 何薇 黄棋 吕红彬 Wu Xiaoqiang;He Wei;Huang Qi;Lyn Hongbin(Department of Ophthalmology,Zigong Third People's Hospital,Zigong 643020,China;Department of Ophthalmology,Chengdu Wenjiang District People's Hospital,Chengdu 6]1130,China;Department of Ophthalmology,Chengdu First People’s Hospital,Chengdu 610071,China;Department of Ophthalmology,Affiliated Hospital of Southwest Medical University,Luzhou 646000,China)

机构地区：[1]四川省自贡市第三人民医院,643020 [2]四川省成都市温江区人民医院眼科,611130 [3]四川省成都市第一人民医院眼科,610071 [4]西南医科大学附属医院眼科,泸州646000

出　　处：《中华眼科医学杂志（电子版）》2019年第6期348-353,共6页Chinese Journal of Ophthalmologic Medicine(Electronic Edition)

基　　金：四川省科技支撑计划项目(2015SZ0086)

摘　　要：目的探讨二项酶诱导剂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Müller细胞凋亡的影响及其与凋亡相关基因B淋巴细胞瘤-2(Bcl-2)和B淋巴细胞瘤-2相关X蛋白(Bax)通路的关系。方法体外培养SD大鼠的Müller细胞,采用数字表法随机分为对照组、高糖组和CPDT干预组。对照组给予25 mM DMEM完全培养基培养;高糖组给予65 mM DMEM完全培养基培养;CPDT干预组给予65 mM DMEM完全培养基和60μM CPDT培养,均培养72 h。采用双染法流式细胞仪检测各组细胞凋亡和细胞周期的情况;采用蛋白免疫印迹杂交技术检测各组细胞Bcl-2和Bax蛋白的表达情况。不同组的组间比较采用单因素方差分析,当差异有统计学意义时,进一步采用LSD法两两比较。结果流式细胞仪检测的结果显示,高糖组的细胞凋亡率为(17.6±3.16)%;对照组的细胞凋亡率为(8.25±0.26)%;CPDT干预组的细胞凋亡率为(13.5±2.25)%,且三组细胞凋亡率间,差异有统计学意义(F=13.48,P<0.05)。蛋白免疫印迹杂交技术检测的结果显示,高糖组Bcl-2蛋白表达量较对照组降低,Bax蛋白表达量较对照组升高,且差异有统计学意义(t=150.38,234.46;P<0.05)。CPDT干预组Bcl-2蛋白表达量较高糖组升高,Bax蛋白表达量较高糖组降低,且差异有统计学意义(t=108.03,36.33;P<0.05)。结论高糖可通过上调Bax,下调Bcl-2表达,降低Bcl-2/Bax的比值,促进细胞凋亡。CPDT可增加高糖影响下的大鼠Müller细胞活性、Bcl-2的表达及Bcl-2/Bax的比值,抑制Bax的表达和高糖引起的大鼠Müller细胞的凋亡,从而起到保护高糖环境中Müller细胞受损伤的作用。Objective This aim of this study was to investigate the effect of 5,6-dihydrocyclopentene-1,2-dithiol-3-thione,a two-enzyme inducer(CPDT)on the apoptosis of rat Müller cells in the high glucose environment and whether it exerts its role through the apoptosis-related genes Bcl-2/Bax pathways,in order to explore the prospect of CPDT in the prevention and treatment of diabetic retinopathy.Methods SD rat Müller cells were cultured in vitro and randomly divided into 3 groups:control group(25 mM DMEM),high glucose group(65 mM DMEM),CPDT intervention group(65 mM DMEM+60 uM CPDT).These cells were cultured for 72 h.Apoptosis and cell cycle changes were detected by the flow cytometry with double staining.The expressions of Bcl-2 and Bax proteins in each group were detected by Western blotting.Data were collected and one-way ANOVA was used to performstatistically analyzed between groups.Results The results of double staining flow cytometry showed that the apoptotic rate of high glucose group was(17.6±3.16%),that of control group was(8.25±0.26%),and that of CPDT intervention group was(13.5%±2.25%).There was statistically significant among them(F=13.48,P<0.05).The results of Western blotting hybridization detection showed that compared with the control group,the expression of Bcl-2 protein in the high glucose group was significantly decreased and the expression of Bax protein was significantly increased(t=150.38,234.46;P<0.05).Compared with the high glucose group,the expression of Bcl-2 protein in the CPDT intervention group increased,Bax protein expression decreased(t=108.03,36.33;P<0.05).Conclusion High glucose could up-regulate Bax,down-regulate Bcl-2 expression,decrease Bcl-2/Bax ratio,and promote apoptosis.CPDT could increase the activity of rat Müller cells,Bcl-2 expression and Bcl-2/Bax ratio,inhibit the expression of Bax from the pressure of high glucose,thereby inhibiting the apoptosis of rat Müller cells induced by high glucose,and had a protective effect on Müller cells in a high glucose environment.

关 键 词：糖尿病性视网膜病变 MULLER细胞 CPDT BCL-2/BAX 凋亡 

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