文章摘要
杨哲 刘平 王嘉翔 单多 车雯 张丽娟.EGCG对H2O2诱导HLE细胞氧化损伤的保护作用[J].,2016,16(28):5410-5413
EGCG对H2O2诱导HLE细胞氧化损伤的保护作用
Protective Effects of EGCG on H2O2-induced Oxidative Damage in HLECells
  
DOI:
中文关键词: EGCG  HLE细胞  caspses-3  caspase-9
英文关键词: EGCG  HLE cells  Caspses-3  Caspase-9
基金项目:国家自然科学基金项目(30973275)
作者单位
杨哲 刘平 王嘉翔 单多 车雯 张丽娟 哈尔滨医科大学附属第一医院眼科三病房 
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中文摘要:
      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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