【摘要】 目的研究P38有丝分裂素激活蛋白激酶(MAPK)在H2O2诱导的人晶状体上皮细胞热休克蛋白质27(HSP27)表达中的作用。方法使用100和200μmol/LH2O2分别以不同时间刺激培养的人晶状体上皮细胞系B3,在阻断实验中加入特异性P38MAPK阻断剂SB203580阻断P38MAPK信号转导通路。采用逆转录聚合酶链反应检测刺激不同时间的人晶状体上皮细胞HSP27mRNA的表达情况,采用免疫印迹法(Westernblotting)检测细胞中HSP27和磷酸化P38MAPK的表达情况。结果(1)H2O2刺激后的人晶状体上皮细胞中HSP27mRNA及其蛋白的表达显著增加;(2)100和200μmol/LH2O2刺激30min,人晶状体上皮细胞应激活化蛋白激酶的活性(平均灰度值)分别增强至800±081和825±050,6h后恢复至基线水平(P<001);磷酸化P38MAPK的表达随H2O2浓度的增高而增加,且在H2O2刺激后15min达到峰值,随后逐渐下降;(3)加用特异性P38MAPK阻断剂SB203580预处理后,200μmol/LH2O2刺激6h的人晶状体上皮细胞HSP27表达(平均灰度值)从3600±082降至875±126,受到显著抑制(P<001)。结论P38MAPK信号转导途径参与H2O2诱导的人晶状体上皮细胞HSP27的表达。(中华眼科杂志,2005,414751)更多还原

【Abstract】 Objective To investigate the role of P38MAPK signaling pathway in hydrogen peroxide （H₂O₂） induced the expression of heat shock protein 27 in human lens epithelial cell line HLE-B3. Methods Human lens epithelial cell line HLE-B3 was cultured and then co-incubated with H₂O₂ （100 μmol/L,200 μmol/L） for different time with or without pretreatment with Specific P38MAPK inhibitor SB203580. RT-PCR and Western blotting were used to detect HSP_<sup>27 mRNA and protein expressions at different time points after H₂O₂ stimulation in human lens epithelial cells. Western blotting was used to detect the expression of phosphorylated P38MAPK. Results The expression of HSP_<sup>27 mRNA and protein was increased significantly after the H₂O₂ stimulation.The activity of stress-activated protein kinase was increased at 30 minutes after treatment with H₂O₂ and decreased to base level at 6 hours （P<0.01）.The expression of phosphorylation of P38MAPK was increased after treatment of H₂O₂ and reached the highest level at 15 minutes after stimulation and then decreased.The expression of HSP_<sup>27 was down-regulated with the pretreatment of SB203580 compared with that in the H₂O₂ group （P<0.01）. Conclusions H₂O₂ induces the expression of heat shock protein 27 in human lens epithelial cell line. The effects are mediated, at least in part, through the activation of P38MAPK signaling pathway.更多还原