复方水蛭滴眼液抑制大鼠晶状体上皮细胞凋亡及其对Bcl-2和Bax基因的调控

目的:探讨复方水蛭滴眼液(compound leech eye drops,Co-SZ)对H_2O_2诱导的大鼠晶状体上皮细胞(lens epithelial cell,LEC)凋亡的抑制作用及其对凋亡相关基因Bcl-2和Bax表达的调控,为将Co-SZ作为防治白内障的有效药物提供科学的依据。方法:采用H_2O_2复制SD大鼠LEC凋亡模型,同时采用Co-SZ干预。置二氧化碳培养箱共同孵育24h后,以末端脱氧核苷酸转移酶介导的生物素化脱脲苷三磷酸末端标记法(terminal deoxynucleotidyl transferase mediated biotin-dUTP nick end labeling,TUNEL)检测LEC凋亡率;透射电镜观察LEC超徵结构改变及凋亡形成;免疫组化法检测凋亡相关基因Bcl-2和Bax的蛋白表达并进行比较。结果:TUNEL法检测结果显示Co-SZ组LEC凋亡率显著低于H_2O_2组。Co-SZ组LEC超微结构凋亡变化也显著轻于H_2O_2组。与H_2O_2组比较,Co-SZ可明显上调Bcl-2表达,下调Bax表达。结论:Co-SZ能育效抑制H_2O_2诱导的LEC凋亡。调控凋亡相关基因Bcl-2和Bax的表达可能是其抑制LEC凋亡的分子机制。

Inhibition effects of compound leech eye drops on apoptosis of lens epithelial cells and expressions of Bcl-2 and Bax genes in rats

Objective:To investigate the inhibition effects of compound leech eye drops(Co-SZ)on apoptosis of lens epithelial cells(LECs)induced by hydrogen peroxide(H_2O_2)and the expressions of apoptosis-related genes Bcl-2 and Bax in rats. Methods:All fresh transparent LECs in SD rats were bathed in culture medium with H_2O_2 in vitro,meanwhile Co-SZ were added in the culture medium.All LECs were incubated for 24 hours.The apoptosis rate of LECs was determined by terminal deoxynucleotidyl transferase mediated biotin-dUTP nick end labeling method (TUNEL).The changes of LEC ultrastructure and the formation of apoptotic body were observed by transmission electron microscopy.The expressions of apoptosis-related genes Bcl-2 and Bax were detected by streptavdin-peroxidase-biotin method. Results:The apoptosis rate of LECs in the Co-SZ-treated group was significantly lower than that in the H_2O_2- treated group.The changes of apoptotic LEC ultrastructure in the Co-SZ-treated group were less than those in the H_2O_2-treated group.The expression of Bcl-2 protein was up-regulated and the expression of Bax protein was down-regulated in the Co-SZ-treated group as compared with the H_2O_2-treated group. Conclusion:The LEC apoptosis induced by H_2O_2 can be inhibited by Co-SZ.The molecular mechanisms of Co-SZ in inhibiting LEC apoptosis may be related to regulating the expressions of apoptosis-related genes Bcl-2 and Bax.