蛋白激酶C_(-α)在晶状体上皮细胞中的表达及其与白内障关系的研究

The expression of protein kinase C-a in lens epithelial cells and its role in cataract formation

目的 研究蛋白激酶C-α(PKC-α)在晶状体上皮细胞中的表达及其在皮质性白内障形成过程中的变化及作用。方法 离体培养的大鼠晶状体,应用Western-blot检测蛋白激酶C在晶状体上皮细胞中的表达和分布;H_2O_2诱导大鼠皮质性白内障形成,Western-blot检测晶状体上皮细胞内蛋白激酶C-α的蛋白表达变化;RT-PCR检测其mRNA含量改变。同时,流式细胞仪检测晶状体上皮细胞调亡率。结果 在正常晶状体上皮细胞中PKC-α高表达,经H_2O_2处理产生凋亡的晶状体上皮细胞中PKC-α的蛋白表达和mRNA含量均降低,与对照组比较,差异有显著意义(P<0.05)。结论 H_2O_2可抑制大鼠晶状体上皮细胞中PKC-α的活性,提示PKC-α与白内障的发生发展有关。

Objective To investigate the expression and role of protein kinase C in rat lens epithelial cells (LECs) with hydrogen peroxide ( H2 02 ) -induced cataract. Methods Rat lenses were cultured in vitro and cataract was induced by 2 mmol/L H2 02 exposure. The expression of protein kinase C in rat LECs during cataract formation was analyzed by Western-blot analysis. RT-PCR was used to detected the changes in mRNA expression. The apoptosis percentage was measured by flow cytometer. Results High protein level of PKC-a was observed in normal lens epithelial cells, but its protein and mRNA concentration were significantly decreased after the H202 treatment in comparison with control group (P <0. 05). Conclusion The expression of protein kinase-a protein and mRNA in lens epithelial cells is regulated by H2 02, suggesting an interaction between protein kinase-a and the formation of cataract.