羽扇豆醇-3β-琥珀酸酯通过调控线粒体凋亡途径诱导HepG2细胞凋亡  

Lupinol-3β-succinate induce apoptosis of HepG2 cells by regulating the mitochondrial apoptosis pathway

作　　者：卜明 王璐 罗然 徐天赐 林宇[3] 葛鹏玲[2] 刘吉成 BU Ming;WANG Lu;LUO Ran;XU Tianci;LIN Yu;GE Pengling;LIU Jicheng(Post-Doctoral Research Center,Qiqihar Institute of Medical Sciences,Qiqihar 161006,China;Mobile Postdoctoral Center,Heilongjiang University of Traditional Chinese Medicine,Harbin 150040,China;College of Pharmacy,Qiqihar Medical University,Qiqihar 161006,China)

机构地区：[1]齐齐哈尔医药科学研究所博士后科研工作站,黑龙江齐齐哈尔161006 [2]黑龙江中医药大学博士后流动站,黑龙江哈尔滨150040 [3]齐齐哈尔医学院药学院,黑龙江齐齐哈尔161006

出　　处：《现代药物与临床》2024年第3期549-556,共8页Drugs & Clinic

基　　金：中央支持地方高校改革发展资金人才培养项目(21B024);齐齐哈尔医学科学院科研项目(QMSI2022Z-01)。

摘　　要：目的探究羽扇豆醇-3β-琥珀酸酯对HepG2细胞凋亡的作用及机制。方法采用0、2.5、5、10、20、40、80μmol/L的羽扇豆醇-3β-琥珀酸酯作用于HepG2细胞24、48、72 h,采用CCK-8法检测各组细胞的增殖活性。采用0、4、8、16μmol/L的羽扇豆醇-3β-琥珀酸酯作用于HepG2细胞48 h后,AO/EB染色法和Annexin V-FITC/PI双染法检测各组细胞的凋亡率;JC-1染色法观察HepG2细胞的线粒体膜电位变化;DCFH-DA染色法观察细胞的活性氧(ROS)水平。Western blotting法检测各组细胞线粒体凋亡途径相关蛋白表达变化。结果羽扇豆醇-3β-琥珀酸酯呈时间和浓度相关性地抑制HepG2细胞增殖(P<0.01)。与对照组相比,羽扇豆醇-3β-琥珀酸酯组细胞的ROS水平、总凋亡率、Bcl-2相关X蛋白(Bax)、细胞色素C、裂解半胱氨酸蛋白酶9(cleaved-Caspase-9)、cleaved Caspase-3蛋白水平呈浓度相关性升高(P<0.01、0.001);线粒体的膜电位、Bcl-2蛋白表达则显著降低(P<0.01)。结论羽扇豆醇-3β-琥珀酸酯可能通过调控线粒体凋亡途径诱导人肝癌HepG2细胞凋亡,进而发挥抗肝癌作用。Objective To investigate the effect of lupinol-3β-succinate on apoptosis of HepG2 cells and mechanism.Methods HepG2 cells were treated with lupinol-3β-succinate(0,2.5,5,10,20,40 and 80μmol/L)for 24,48,and 72 h,and the proliferation activity of the cells was detected by CCK-8 method.HepG2 cells were treated with lupinol-3β-succinate(0,4,8,and 16μmol/L)for 48 h,and the apoptosis rate of each group was detected by AO/EB staining and Annexin V-FITC/PI double staining.Changes of mitochondrial membrane potential in HepG2 cells were observed by JC-1 staining.Levels of ROS were observed by DCFH-DA staining.Western blotting method was used to detect the changes of mitochondrial apoptosis-related protein expression in each group.Results Lupinol-3β-succinate showed significant time and dose dependent inhibition of HepG2 cell proliferation(P<0.01).Compared with control group,ROS level,total apoptosis rate,Bax,Cyt-C,cleaved Caspase-9,and cleaved Caspase-3 protein levels were increased in a dose�dependent manner after lupinol-3β-succinate treatment(P<0.01,0.001).Mitochondrial membrane potential and Bcl-2 protein expression were significantly decreased(P<0.01).Conclusion Lupinol-3β-succinate may induce apoptosis of human hepatoma HepG2 cells by regulating mitochondrial apoptosis pathway,and finally play an anti-hepatocellular carcinoma role.

关 键 词：羽扇豆醇-3β-琥珀酸酯 肝癌 细胞凋亡 活性氧 线粒体 Bcl-2相关X蛋白 细胞色素C 半胱氨酸蛋白酶9 

分 类 号：R285[医药卫生—中药学]