17β-雌二醇对高同型半胱氨酸诱导人成骨肉瘤MG63细胞损伤的保护作用及机制  

Protection effects and mechanism of 17β-estradiol in high homocysteine induced injury of human osteosarcoma MG63 cells in vitro

作　　者：陈学秋[1,2] 戴纪男[1] 刘承杏[1] 边立功[1] 郭家智[1] 孙俊[1] 桂莉 陆地[1] 

机构地区：[1]昆明医学院人体解剖学教研室,昆明650500 [2]云南省中医学院人体解剖学教研室,昆明650500 [3]云南省第三人民医院内分泌科,昆明650011

出　　处：《解剖学报》2011年第5期635-639,共5页Acta Anatomica Sinica

基　　金：云南省应用基础研究重点资助项目(2008CC007);云南省中青年学术和技术带头人后备人才培养资助项目(2009CI033)

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。Objective To explore the protection effects and molecular mechanisms of 17βestradiol （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-E2 significantly 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 HHey and down-regulated the expression of Caspase-3 and Bax. Conclusion 17β-E2 can protect MG63 cells from HHey induced oxidative injury by suppressing the production of ROS and the down-regulation of Caspase-3 and Bax.

关 键 词：MG63细胞 17Β-雌二醇 高同型半胱氨酸 活性氧簇 荧光探针 Hoechst 33342/PI核双染法 免疫印迹法 

分 类 号：R322.7[医药卫生—人体解剖和组织胚胎学]