GRIM-19在H_2O_2诱导的晶状体上皮细胞氧化应激中的表达

Research of the Expression of Controlling Genes of Cell Apoptosis GRIM-19 in Human Lens Epithelial Cells Induced by Hydrogen Peroxide

目的观察干扰素/维甲酸联合应用诱导细胞凋亡相关基因19(GRIM-19)在不同浓度H2O2诱导的人晶状体上皮细胞(HLEC)氧化应激中的表达变化。方法免疫荧光法检测GRIM-19在正常HLEC中的表达及定位;建立HLEC氧化损伤模型:分别用浓度为0、20、100、300、500和800μmol/L的H2O2处理HLEC 1 h后换液,继续孵育6、24 h;MTT法检测各浓度H2O2对HLEC的生长抑制作用;Western blot法检测6 h组HLEC中GRIM-19表达的变化。结果免疫荧光结果示:GRIM-19主要表达于HLEC细胞质中;MTT结果示:100~800μmol/L H2O2对HLEC有增殖抑制作用,且呈剂量依赖关系,与对照组相比差异有统计学意义(P均<0.01),20μmol/L组与空白对照组未见明显差异(P>0.05);Western blot结果显示:GRIM-19在各浓度组中相对表达量分别为1.466±0.018,1.509±0.038,1.599±0.014,1.709±0.058,1.813±0.010,1.583±0.021,实验组均显著高于空白对照组(P<0.05),正常对照组与空白对照组未见明显差异。结论正常HLEC中存在GRIM-19表达。在一定浓度H2O2诱导的氧化应激中,GRIM-19的表达显著增加。

Objective To investigate the effects of hydrogen peroxide （H202） at various concentrations on the proliferation and GRIM-19 expres- sion of human lens epithelial cells （HLEC）. Methods The expression and position of GRIM-19 was detected by immunofluorescence. The oxida- tion stress models of HLEC were established by different dose of H202 （0,20,100,300,500,800 b^mol/L） on HLEC, The survival and proliferation of the cells was quantified by MTF assay before treatment and at 6,24 hours after one hour treatment of H202. The concentration of GRIM- 19 in the HLEC was measured by Western blot at 6 hours after the treatment of H202. The immunofluorescence results shows the expression of GRIM- 19 in LEC. It mainly exists in cytoplasm. Results The immunofluorescence showed the expression of GRIM- 19 in LEC. It mainly existed in cytoplasm, Results of MTF show that a certain concentration range of H202 has a proliferation inhibition on HLEC. There is a dose-response relationship between 100 and 800 txmol/L. Compared with the control, difference have statistical significance （F6h =146.708, P 〈0.01 ; Fah =623.859, P 〈 0.01 ）. We did not see an obvious difference between the group of 20μmol/L and the blank control （P 〉 0.05 ） .The relative expression quantity of GRIM- 19 detect- ed by Western blot under 0, 20,100, 300, 500, 800 p^mol/L treatment separately, was 1.466±0.018,1.509±0.038, 1.599±0.014, 1.709±0.058, 1.813±0.010, 1.583±0.021, which were significantly higher than the control group, excepted the normal group （F =53.732, P 〈 0.05 ）. Conclusion It was confirmed that the GRIM- 19 expressed in HLEC. There was a significantly increase of GRIM- 19 in HLEC under the oxidative stress of a cer- tain concentration of H202.