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氧化苦参碱诱导人肝癌HepG2细胞凋亡的可能机制
侯颖1△,曹蔚1△,李涛2,刘水冰1,张晓楠1,李旭波1,田琼1*
0
(1.第四军医大学药学系药理学教研室,西安 710032;2.西安交通大学医学院法医系,西安 710061)
摘要:
目的:观察氧化苦参碱(OM)对人肝癌HepG2细胞凋亡的影响,探讨其可能的作用机制。方法:用不同浓度的OM处理HepG2细胞,MTT法检测OM对HepG2细胞生长增殖的抑制作用;应用Hochest荧光染色法观察OM处理后细胞形态的变化;流式细胞术(FCM)检测细胞凋亡率;免疫印迹法检测OM处理后细胞凋亡相关蛋白磷酸化Akt、caspase-3、Bax、Bcl-2和Bcl-xL的表达。结果:OM抑制HepG2细胞的增殖并呈一定的量效和时效关系;HepG2细胞与OM作用后出现典型的凋亡细胞形态改变,细胞凋亡率增高;OM处理HepG2细胞后,磷酸化Akt蛋白表达下降,caspase-3可被蛋白酶水解形成相对分子质量17 000活性片段,Bax蛋白表达增强,Bcl-2和Bcl-xL蛋白表达下调。结论:OM可诱导HepG2细胞凋亡,可能与其调节PI3K/Akt信号通路,抑制Bcl-2和Bcl-xL的活性,激活caspase-3有关。
关键词:  氧化苦参碱  实验性肝肿瘤  细胞凋亡
DOI:10.3724/SP.J.1008.2008.00634
投稿时间:2008-03-01修订日期:2008-05-04
基金项目:
Oxymatrine-induced apoptosis of HepG2 cells:the possible mechanism
HOU Ying1△,CAO Wei1△,LI Tao2,LIU Shui-bing1,ZHANG Xiao-nan1,LI Xu-bo1,TIAN Qiong1*
(1.Department of Pharmacology,School of Pharmacy,the Fourth Military Medical University,Xi’an 710032,China;2.Department of Forensic Medicine,Medical School of Xi’an Jiaotong University,Xi’an 710061)
Abstract:
Objective:To investigate the effects of oxymatrine(OM) on the apoptosis of human hepatoma HepG2 cells and its possible mechanisms.Methods: HepG2 cells were treated with different concentrations of OM.The proliferation inhibition was measured by MTT assay and the apoptosis of HepG2 cells were examined by Hochest staining method.Flow cytometry was used to analyze the cell cycle distribution and apoptosis rate. The expression of caspase-3,Bcl-2,Bcl-xL and Bax proteins was assayed by Western blotting assay.Results: OM inhibited HepG2 cells growth in a time- and dose-dependent manner.After treatment with OM for 24 hours,some cells appeared typical apoptotic characteristics and the apoptosis rate was increased.Treatment with OM also increased caspase-3 activity and Bax expression in HepG2 cells,and decreased the expression levels of Bcl-2 and Bcl-xL.Conclusion: OM can induce HepG2 cell apoptosis,which may be related to the down-regulation of PI3K/Akt signal pathway,suppression of Bcl-2 and Bcl-xL activity,and activation of caspase-3.
Key words:  oxymatrine  experimental liver neoplasms  apoptosis