干扰素／维甲酸诱导细胞凋亡相关基因19表达与紫外线诱导的人晶状体上皮细胞凋亡的关系  

Effects of gene associated with retinoid-interferon-induced mortality-19 on ultraviolet A radiation-induced human lens epithelial cell apoptosis in vitro

作　　者：王思敏[1] 冯浩[1] 林婷婷[1] 刘加勇[1] 宁宏[1] 

机构地区：[1]中国医科大学附属第一医院眼科,沈阳110001

出　　处：《中华实验眼科杂志》2013年第4期358-361,共4页Chinese Journal Of Experimental Ophthalmology

摘　　要：背景长期暴露于紫外线照射的环境中可通过破坏细胞线粒体的跨膜电位等机制促进相应组织中的细胞凋亡，包括人晶状体上皮细胞（LECs）。干扰素／维甲酸诱导细胞凋亡相关基因-19（GRIM-19）位于线粒体呼吸链Ⅰ上，与紫外线照射诱导的LECs凋亡过程是否有关尚不明确。目的探讨GRIM-19与长波紫外线照射诱导的人LECs损伤的关系。方法将人LECs系SRA01／04用常规培养方法在含质量分数10％胎牛血清的α-MEM培养液中进行培养，当细胞生长至对数生长期并达到80％融合时用长波紫外线照射，时间分别为5、10、15、20、25min，长波紫外线剂量分别为30、60、90、120、150mJ／cm^2，照射后继续培养1h，对照组细胞与实验组培养方法相同，但不用长波紫外线照射。采用流式细胞技术测定各组培养的凋亡率，应用Westernblot法检测各组培养上清液中GRIM-19蛋白的表达强度和相对表达量，采用单因素方差分析法对不同组间LECs凋亡率和GRIM-19蛋白相对表达量的差异进行比较，用Pearson积矩线性相关分析法评估LECs凋亡率与GRIM-19蛋白相对表达量的关系。结果不同剂量的紫外线照射培养的LECs后各组细胞凋亡率的总体比较差异有统计学意义（F=149．32，P〈0．01），其中60、90、120及150mJ／em。紫外线照射组与对照组比较，细胞凋亡率均明显增加，差异均有统计学意义（q=-17．02、-25．20、-29．41、-8．61，P〈0．01）。不同剂量的紫外线照射培养的LECs后细胞上清液中GRIM·19的表达明显增强，各组LECs上清液中GRIM-19的相对表达量（GRIM-19／β-actin）的总体差异有统计学意义（F=6．87，P〈0．05），其中60、90、120、150mJ／cm。紫外线照射组细胞上清液中GRIM-19的相对表达量分别为2．01±O．76、2．98±1．80、3．97±1．61和2．42±1．28，均明显高于对照组的0．56±0．23，差异均有统计学意义（q=-4．12�Background Ultraviolet irradiation promotes cellular apoptosis by affecting the mitochondrial transmembrane potential, including human lens epithelial cells （LECs）. Gene associated with retinoid-interferon- induced mortality-19 （GRIM-19）, a cell death regulatory protein, is essential for the assembly and function of mitochondrial complex I. However,whether LECs apoptosis induced by ultraviolet irradiation is related to GRIM-19 is still unclear. Objective The purpose of this study was to investigate the relationship between the apoptosis of human LECs caused by ultraviolet with GRIM-19 expression in vitro. Methods Human LEC line（SRA01/04）was cultured in α-MEM containing 10% fetal bovine serum. The cells were exposed to ultraviolet ray at doses of 0,30,60, 90,120 or 150 mJ/cm^2 when cell growth reached the logarithmic phase and 80% confluency. The rate of apoptosis of the ceils was assayed using flow cytometry, and the level of expression and relative amount of GRIM-19 protein （GRIM-19/β-actin） were detected by Western blot. The relationship between apoptosis and the GRIM-19/β-actinvalue among the different treatment groups was compared using One-way ANOVA, and the correlation of LECs apoptosis rate and GRIM-19 expression level was assessed by Pearson linear analysis. Results A significant difference was found in the apoptosis rate among the different treatment groups（ F= 149.32 ,P〈O. 01 ）. Compared with the 0 mJ/cm2 ultraviolet irradiation group, the apoptosis rate of LECs was significantly increased in the 60,90,120 and 150 mJ/cm2 ultraviolet irradiation groups （ q = 17.02, - 25.20, - 29.41, - 8.61, P 〈 0.01 ）. The expression of the GRIM-19 protein in the LECs suspension was enhanced by ultraviolet irradiation at 60,90,120 and 150 mJ/cm2. The relative expression of the GRIM-19 protein （GRIM-19/^-actin）was significantly different among the various groups （F= 6.87 ,P〈0. 05 ） , and the GRIM-19/^-actin values in the 60,90,120,150 mJ/cm~ ultraviolet irradiation groups were elev

关 键 词：干扰素 维甲酸诱导细胞凋亡相关基因19 人晶状体上皮细胞 长波紫外线 细胞凋亡 

分 类 号：R965[医药卫生—药理学]