PP242对人晶状体上皮细胞增殖的抑制作用及GRIM-19表达的影响  

The Influence of PP242 on the Inhibition of the Human Lens Epithelial Cell Proliferation and the Expression of GRIM-19

作　　者：冯浩[1] 林婷婷[1] 王思敏[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：《中国医科大学学报》2013年第5期416-419,424,共5页Journal of China Medical University

基　　金：辽宁省科学技术计划(2008225016)

摘　　要：目的观察PP242对体外培养的人晶状体上皮细胞(HLEC)增殖和GRIM-19表达的影响。方法对永生系人晶状体上皮细胞SRA01/04进行培养及传代,Cell Counting Kit-8比色法检测PP242(100,250,500,750,1 000 nmol/L)处理细胞株SRA01/04后12,24,48 h细胞生长情况;应用免疫印迹法及实时定量RT-PCR检测使用PP242(100,250,500 nmol/L)培养细胞48 h后GRIM-19及其mRNA的表达情况。结果 100,250,500,750,1 000 nmol/L PP242对人晶状体上皮细胞系SRA01/04作用12,24,48 h后,其抑制率12 h组分别为8.59%,14.53%,19.62%,23.01%,28.80%;24 h组分别为10.22%,19.67%,34.92%,42.90%,46.86%;48 h组分别为24.19%,48.13%,54.54%,61.36%,65.34%,与对照组比较差异均有统计学意义(P<0.05);100,250,500 nmol/L PP242处理人晶状体上皮细胞48 h后,免疫印迹法检测结果显示GRIM-19蛋白的表达水平逐渐增强;实时定量RT-PCR检测结果显示,GRIM-19 mRNA表达增强,绝对定量mRNA结果分别为0.700±0.112,0.994±0.128,1.584±0.173,与正常对照组(0.456±0.177)比较差异有统计学意义(P<0.05)。结论 PP242对体外培养的人晶状体上皮细胞的增殖存在抑制作用,其作用强度呈浓度和时间依赖性;人晶状体上皮细胞中存在GRIM-19的表达;PP242在抑制人晶状体上皮细胞的增殖过程中可上调GRIM-19的表达。Objective To investigate the influence of PP242on the proliferation of human lens epithelial cells （HLEC） and the expression of GRIM-19. Methods hnmortal HLEC SRA01/04 were cultured and divided into five groups and treated with different concentrations of PP242 （ 100,250,500,750, 1 000 nmol/L） respectively. With Cell Counting Kit-8 Calorimetric assay,the growth of ceils in each group was examined at 12 h,24 h and 48 h after treatment. By using real-time quantitative polymerasechain reaction and inmmnoblot assay,the GRIM- 19 expression of the cultured cells SRA01/04were examined 48 h after PP242 treatment at both mRNA and protein levels. Results The in- hibition ratio of different concentration of PP242 on culturing HLEC was as below： 12 h：8.59%, 14.53%, 19.62%, 23.01%, 28.80%;24 h： 10.22%, 19.67%, 34.92%, 42.90%, 46.86% ; 48 h ： 24.19%, 48.13%, 54.54%, 61.36%, 65.34%. Compared with the control group, the dif- ference was of statistical significance （P 〈 0.05 ）. According to the immunoblot assay ,the expressing level of GRIM-19 protein in HLEC was significantly increased after 48 h treatment of PP242;the real-time quantitative PCR result confirmed that the expression of GRIM-19 mRNA was enhanced. The absolute quantitative result of mRNA was 0.700±0.112,0.994±0.128, and 1.584±0.173, respectively. Compared with the control group （0.456±0.177）, the difference was statistically significant （P 〈 0.05 ）. Conclusion PP242 showed an concentration and time dependent inhibitive effect on in vitro cultured HLEC,PP242 up-regulated the expression of GRIM-19 in the process of inhibiting the prolif- eration of HLEC.

关 键 词：PP242 人晶状体上皮细胞 GRIM-19 

分 类 号：R776.1[医药卫生—眼科]