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[摘要]
目的: 探讨17-β雌二醇对高糖诱导人视网膜色素上皮(retinal pigment epithelial,RPE)细胞的保护作用和可能机制。
方法:RPE细胞传代培养,随机对照法分为四组:空白对照组:5.5mmol/L葡萄糖常规培养基进行处理; 高糖组:100mmol/L葡萄糖作用12h; 17-β雌二醇低浓度组:10μmol/L 17-β雌二醇孵育24h后,加入100mmol/L葡萄糖作用12h; 17-β雌二醇高浓度组:100μmol/L 17-β雌二醇孵育24h后,加入100mmol/L葡萄糖作用12h。MTT比色法检测细胞活力,Hochest33258染色观察细胞凋亡,H2DCFDA染色检测细胞内活性氧(reactive oxygen species,ROS)水平,比色法检测过氧化氢酶(catalase,CAT)、超氧化物歧化酶(superoxide dismutase,SOD)和丙二醛(malondialdehyde,MDA)的表达。
结果:随着葡萄糖浓度的增加,RPE细胞活性逐渐下降,17-β雌二醇能明显抑制高糖诱导的RPE细胞活力的下降,降低RPE细胞凋亡率,抑制细胞内ROS生成。此外,17-β雌二醇能明显增加RPE细胞内CAT、SOD表达,降低MDA的表达。
结论:17-β雌二醇能有效抑制高糖对RPE细胞的损伤,从而为用于治疗视网膜相关疾病提供可靠的实验依据。
[Key word]
[Abstract]
AIM: To discuss the protective effects and possible mechanisms of 17β-estradiol on human retinal pigment epithelial(RPE)cells induced by high glucose.
METHODS: RPE cells were cultured and divided into four groups according to randomized controlled method: blank control group: the cells were treated with 5.5mmol/L routine glucose medium for processing; high glucose group: cells were treated with 100mmol/L glucose for 12h; 17β-estradiol low concentration group: after treated with 10 μmol/L 17β-estradiol, cells were treated with 100mmol/L glucose for 12h; 17β-estradiol high concentration group: after treated with 100 μmol/L 17β-estradiol, cells were treated with 100mmol/L glucose for 12h. Cell viability were tested by MTT colorimetric detection. Cells apoptosis were detected by Hochest33258 staining. Intracellular reactive oxygen species(ROS)level were detected by H2DCFDA staining. Expression of CAT, SOD and MDA were tested by colorimetric detection.
RESULTS: RPE cell activity decreased with the concentration of glucose increased; 17β-estradiol inhibited high glucose-induced cell viability decrease in RPE cells, decreased the apoptosis rate of RPE cells and intracellular ROS generation; besides, 17β-estradiol significantly increased the expression of CAT, SOD and decreased the expression of MDA in RPE cells.
CONCLUSION: The 17β-estradiol effectively inhibited high glucose -induced RPE cells damage, which provide reliable experimental basis for the treatment of injuries in RPE cells.
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