龙葵碱调控Bcl-2与Bax蛋白表达及caspase-3活性诱导HepG2细胞凋亡的研究  

Induction of solanine on HepG2 cell apoptosis by regulation of Bcl-2/Bax expression and caspase-3 activity

作　　者：高世勇[1,2] 徐丽丽[1,2] 季宇彬[1,2] 

机构地区：[1]哈尔滨商业大学生命科学与环境科学研究中心药物研究所博士后科研工作站,黑龙江哈尔滨150076 [2]国家教育部抗肿瘤天然药物工程研究中心,黑龙江哈尔滨150076

出　　处：《中草药》2009年第10期1607-1612,共6页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(30400591);黑龙江省博士后基金资助项目;教育部博士点基金项目(200802400001);哈尔滨市青年基金资助项目(2004AFQXJ035)

摘　　要：目的探讨龙葵碱诱导HepG2细胞凋亡的作用机制。方法透射电镜观察凋亡细胞形态变化,原位缺口末端检测法(TUNEL法)检测DNA断裂情况,流式细胞术检测细胞凋亡率,间接免疫荧光法激光共聚焦扫描显微术检测Bcl-2与Bax蛋白表达,比色法检测caspase-3活性的变化。结果在透射电镜下观察,龙葵碱组细胞出现细胞固缩,染色质致密,核凝聚固缩,染色体断裂形成核碎块,凋亡小体形成等细胞凋亡特征形态。TUNEL法发现龙葵碱高、中、低剂量组HepG2细胞均有绿色荧光,阴性对照组无荧光。流式细胞术分析表明0.4、2、10μmol/L龙葵碱作用HepG2细胞24h凋亡率分别为4.0%、8.5%、20.1%。同时,龙葵碱升高caspase-3活性,下调Bcl-2蛋白表达,上调Bax蛋白表达。结论龙葵碱通过降低Bcl-2/Bax的值,激活caspase-3酶活性诱导HepG2细胞凋亡。Objective To study the mechanism of solanine on apoptosis in HepG2 cell. Methods Morphology of apoptosis was observed by electron microscope; TUNEL Assay was adopted to observe the DNA cleavage of apoptotic cells, apoptosis rate was analyzed by flow cytomety. Indirect immunofluorescence assay was adopted to study the Bcl-2/Bax protein by Laser Confocal Scan Microscope. Relative caspase-3 activity was determined by colorimetric assay. Results Characteristic morphology of apoptosis, such as shrinkage, ehromatin dense, nuclear condensation pool, the nuclear fragments, and apoptotic bodies, etc. was found. By TUNEL assay, DNA fragmentation was found in solanine groups, but none in control group. The early apoptosis rates induced by 0. 4, 2, and 10 mol/L solanine were 4.0%, 8.5%, and 20.1 %, respectively. And solanine increased caspase-3 activity of HepG2 cells and decreased the Bcl-2 protein expression, increased the Bax protein expression. Conclusion Solanine could induce the HepG2 cell apoptosis by decreasing the Bcl-2 protein expression, increasing the Bax protein expression to activate the caspase-3 activity.

关 键 词：龙葵碱 细胞凋亡 CASPASE-3 BCL-2 BAX 

分 类 号：R286.91[医药卫生—中药学]