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目的：探讨表没食子酸酯(EGCG)对体外培养的人晶状体上皮(human lens epithelial, HLE)细胞氧化损伤的保护作用及可能机制。方法：HLE细胞传代培养，分为阴性对照组:以正常培养液培养；氧化损伤组：100 umol·L-1的H2O2作用12 h；EGCG 低浓度组：10 umol·L-1EGCG 孵育24 h后，加入H2O2作用12 h；EGCG高浓度组：100 umol·L-1EGCG孵育24 h后，加入H2O2作用12 h。 MTT 比色法检测细胞活力，流式细胞仪检测细胞凋亡率，Hochest33258 染色观察凋亡细胞形态，比色法检测凋亡相关因子 caspses-3 及caspase-9 的表达。结果：EGCG能明显抑制H2O2诱导的HLE 细胞活力的下降，用不同浓度EGCG处理后，HLE 细胞活性分别提高到51.00%± 2.37%和63.67%± 2.29%，与氧化损伤组(40.33%± 2.86%)比较差异具有统计学意义(P<0.05)；经不同浓度EGCG处理后，HLE细胞凋亡率分别下降至33.33± 3.12%和22.80± 1.67%，与氧化损伤组(43.03± 2.43%)比较差异具有统计学意义(P<0.05)；此外，EGCG 还能明显减少H2O2所致HLE 细胞内caspses-3 及caspase-9 的表达。结论：EGCG 通过抑制 caspses-3 及caspase-9 的表达有效抑制了H2O2对HLE 细胞的损伤，从而为其用于治疗HLE 细胞损伤提供可靠的实验依据。

英文摘要:

Objective:To discuss the protective effects and possible mechanisms of EGCG in vitro cultured human les epithelial (HLE) cells induced by oxidative damage.Methods:HLE cells were subcultured, devided into negative control group: cultured with normal culture medium; oxidative injury group:100 umol·L-1 H2O2 treated for 12 h; EGCG low dose group: 10 umol·L-1 EGCG incubated for 24 h then treated with 100 umol·L-1 H2O2 for 12 h; and EGCG high dose group: 100 umol·L-1EGCG incubated for 24 h then treated with 100 umol·L-1 H2O2 for 12 h. Cell viability were tested by MTT colorimetric detection, apoptosis rate was detected by flow cytometry, apoptotic cell morphology was observed by hochest33258 staining, expression of apoptosis-related factors caspses-3 and caspase-9 were tested by colorimetric detection.Results:EGCG inhibited H2O2-induced cell viability decrease in HLE cells, after treated with different concentrations of EGCG, HLE cells activity increased to 51.00%± 2.37% and 63.67% ± 2.29 %, which has statistical significance difference compared with oxidative damage group (40.33%± 2.86%)(P<0.05); after treated with different concentrations of EGCG, the apoptosis rate of HLE cells decreased to 33.33± 3.12% and 22.80± 1.67%, respectively, which has statistical significance difference compared with oxidative damage group (43.03 ± 2.43%) (P<0.05); in addition, EGCG also significantly reduced the H2O2-induced expression of caspses-3 and caspase-9 in HLE cells.Conclusion:EGCG effective inhibited H2O2-induced HLE cells damage by inhibiting the expression of caspses-3 and caspase-9, which provide reliable experimental basis for the treatment of injuries in HLE cells.

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