摘要
目的 研究非激素类的雌激素拮抗物Idoxifene与氧化应激导致的鼠肝细胞凋亡之间的关系,进一步阐明其抗鼠肝纤维化机制。方法 用次氮基三乙酸铁 (ferricnitrilotriacetate ,FeNTA)导致培养的鼠肝细胞处于氧化应激状态 ,在idoxifene存在的条件下培养细胞 ,用流式细胞仪 ,酶联免疫测定法 ,Westernblot及凝胶阻滞实验等方法分别检测凋亡细胞 ,caspase-3活性,Bcl-2蛋白家族的表达及结合物蛋白 1(activatorprotein-1,AP-1)结合活性。结果 Idoxifene能显著抑制氧化应激导致的鼠肝细胞凋亡。在鼠肝细胞中 ,氧化应激可降低Bcl-2及Bcl-XL蛋白含量,激活caspase-3与AP-1。Idoxifene与处于氧化应激状态的细胞孵育 12h ,使Bcl-2及Bcl-XL的蛋白含量分别是仅处于氧化应激状态下的3.01倍及1.68倍,使caspase-3活性下降了36.92%。同时idoxifene显著抑制氧化应激诱导的AP-1与其调控位点的结合活性 ,且此AP-1中的主要成分为c-jun与c-fos。结论 Idox ifene能通过抑制Bcl-2及Bcl-XL 蛋白含量的下降及caspase-3及AP-1活性 ,抑制氧化应激导致的鼠肝细胞凋亡。
Abstract
OBJECTIVE To study the effects of the non steroidal estrogen antagonist idoxifene on hepatocyte apoptosis induced by oxidative stress METHODS After hepatocytes were incubated with or without FeNTA(100 mmol·L-1)in the presence or absence of idoxifene(10-9,10-8,10-7mol·L-1 ),apoptotic hepatocytes were examined by flow cytometry .The protein levels of Bcl-2 family,activity of caspase-3 and activator protein-1(AP-1) were detected by western blot,enzyme linked immunosorbent assay and electrophoretic mobility shift assay respectively RESULTS Idoxifene inhibited oxidative stress induced hepatocyte apoptosis.The protein levels of Bcl-2 and Bcl-XL in hepatocytes cultured with FeNTA plus idoxifene were 3.01 and 1.68 times of those in hepatocytes cultured with FeNTA alone respectively. Idoxifene reduced the activity of caspase-3 by 36.92% and significantly inhibited AP-1 activity in hepatocytes undergoing oxidative stress.The activated AP-1 mainly consists of c-jun and c-fos.The activated AP-1 mailnly consists of c-jum and c-fos. CONCLUSION Idoxifene could inhibit oxidative stress-induced hepatocyte apoptosis through maintaining the protein levels of Bcl-2 and Bcl-XL and suppressing the activity of caspase-3 and AP-1.
关键词
idoxifene /
肝纤维化 /
氧化应激 /
凋亡
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Key words
idoxifene /
hepatic fibrosis /
oxidative stress /
apoptosis.
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周亚军;陈宏山;沙宝熙.
Idoxifene抗氧化应激导致的鼠肝细胞凋亡的机制[J]. 中国药学杂志, 2005, 39(04): 278-281
ZHOU Y-jun;CHEN Hong-shn;SH o-xi.
Antiapoptotic mechanisms of idoxifene on cultured rat hepatocytes undergoing oxidative stress [J]. Chinese Pharmaceutical Journal, 2005, 39(04): 278-281
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参考文献
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脚注
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基金
江苏省教育厅自然科学基金资助项目(02KJB310003);教育部留学回国人员基金资助项目(2002247)
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