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

摘要

Objective To investigate the influence of PP242on the proliferation of human lens epithelial cells (HLEC) and the expression of GRIM-19.Methods Immortal 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 Colorimetric assay,the growth of cells in each group was examined at 12 h,24 h and 48 h after treatment.By using real-time quantitative polymerasechain reaction and immunoblot 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 inhibition 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 difference 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 proliferation of HLEC.%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.