【摘要】 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。更多还原

【Abstract】 Objective To explore the protection effects and molecular mechanisms of 17β-estradiol(17β-E2) in high homocysteine(HHcy) induced injury of human osteosarcoma MG63 cells in vitro.Methods Human osteosarcoma MG63 cells cultured in vitro were treated with HHcy in different concentrations at different time after 17β-estradiol intervention treatment.The levels of reactive oxygen species(ROS) were detected with a 2′,7′-dichloro-fluorescin diacetate probe(H2DCFDA).The apoptotic and necrotic cells were determined by double nuclear staining with Hoechst 33342/PI.The expression levels of Caspase-3 and Bax were measured with Western blotting.Results 1.17β-E2significantly suppressed the HHcy induced production of ROS in MG63 cells and the extent of suppression was positively associated with the dose and time.2.17β-E2 significantly prevented MG63 cells from apoptosis or necrosis induced by HHcy and down-regulated the expression of Caspase-3 and Bax.Conclusion 17β-E2 can protect MG63 cells from HHcy induced oxidative injury by suppressing the production of ROS and the down-regulation of Caspase-3 and Bax.更多还原