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目的探讨17β-雌二醇(17β-E2)对高同型半胱氨酸(HHcy)诱导人成骨肉瘤MG63细胞损伤的保护作用及其分子机制。方法体外培养的人成骨肉瘤MG63细胞经17β-E2干预后,再以不同浓度和(或)不同时间的HHcy处理后,采用2’,7’-二氯二氢荧光素二乙酸盐(H2DCFDA)探针检测细胞内活性氧(ROS)水平变化;通过Hoechst 33342/碘化丙啶(PI)核双染法观察计数凋亡和坏死细胞;采用免疫印迹法检测促凋亡相关蛋白Caspase-3和Bax的表达变化。结果 1.17β-E2可明显抑制HHcy诱导的MG63细胞内ROS的增加,抑制效果随剂量和时间而增强;2.17β-E2可明显下调促凋亡相关蛋白Caspase-3和Bax的表达,对抗HHcy诱导的MG63细胞的凋亡和(或)坏死。结论 17β-E2可通过抑制ROS的产生,下调凋亡相关蛋白Caspase-3和Bax的表达而增强MG63细胞抗HHcy所诱导的氧化损伤作用。
Objective To investigate the protective effect of 17β-estradiol (17β-E2) on human osteosarcoma MG63 cells induced by high homocysteine (HHcy) and its molecular mechanism. Methods Human osteosarcoma MG63 cells cultured in vitro were treated with 17β-E2 and treated with HHcy at different concentrations and / or different times. The cells were treated with 2 ’, 7’-dichlorodihydrofluorescein diacetate H2DCFDA) probe was used to detect the level of intracellular reactive oxygen species (ROS). Apoptotic and necrotic cells were counted by Hoechst 33342 / propidium iodide (PI) staining. Western blotting was used to detect the expression of Caspase- 3 and Bax expression changes. 17β-E2 significantly inhibited the increase of ROS in MG63 cells induced by HHcy, and the inhibitory effect was enhanced with dose and time. 2.17β-E2 significantly down-regulated the expression of apoptosis-related proteins Caspase-3 and Bax, Of MG63 cells in apoptosis and / or necrosis. Conclusion 17β-E2 can enhance the anti-HHcy-induced oxidative damage in MG63 cells by inhibiting the production of ROS and down-regulating the expression of apoptosis-related proteins Caspase-3 and Bax.