澳洲茄边碱对人肝癌HepG2细胞生长和凋亡的影响及机制初探  

Preliminary Study on Effects of Solamargine on the Growth and Apoptosis of Human Hepatocarcinoma Cells HepG2 and Its Mechanism

作　　者：吴晶晶 张文娟 张骏鸿 张雅婷 黎莉 卢月 危建安 韩凌 WU Jingjing;ZHANG Wenjuan;ZHANG Junhong;ZHANG Yating;LI Li;LU Yue;WEI Jian’an;HAN Ling(Research Team of Molecular Biology and System Biology of Chinese Medicine,the Second Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510120,China;The Second Clinical College of Guangzhou University of Chinese Medicine,Guangzhou 510120,China;State Key Laboratory of Dampness Syndrome of Chinese Medicine,the Second Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510120,China;Guangdong Provincial Key Laboratory of Clinical Research on Traditional Chinese Medicine Syndrome/Guangdong Clinical Research Center for Dermatosis in Chinese Medicine/Guangdong-Hong Kong-Macao Joint Lab for Chinese Medicine and Immune Disease Research,Guangzhou 510120,China)

机构地区：[1]广州中医药大学第二附属医院中医药分子生物与系统生物学研究团队,广州510120 [2]广州中医药大学第二临床医学院,广州510120 [3]广州中医药大学第二附属医院省部共建中医湿证国家重点实验室,广州510120 [4]广东省中医证候临床研究重点实验室/广东省中医皮肤临床医学研究中心/粤港澳中医药与免疫疾病研究联合实验室,广州510120

出　　处：《中国药房》2021年第24期2963-2969,共7页China Pharmacy

基　　金：国家自然科学基金(青年科学基金)资助项目(No.81902752);广东省科技计划项目(No.2020B1212030006);广东省自然科学基金项目(No.2020A1515010607);省部共建中医湿证国家重点实验室重点项目(No.SZ2021ZZ29);广东省中医院-瑞博奥研究项目(No.YN2018RBA02)。

摘　　要：目的:探讨澳洲茄边碱对人肝癌HepG2细胞生长和凋亡的影响,并研究其潜在作用机制。方法:考察0(空白组)~12μmol/L澳洲茄边碱作用24、48 h对HepG2细胞存活率的影响,0(空白组)、6μmol/L澳洲茄边碱作用10 d对细胞克隆形成的影响,0(空白组)、4、6、8μmol/L澳洲茄边碱作用24 h对细胞凋亡率和细胞中B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)、胱天蛋白酶3(caspase-3)mRNA表达量以及Bcl-2、剪切的caspase-3(cleaved caspase-3)蛋白表达量和磷酸化腺苷一磷酸活化蛋白激酶α(p-AMPKα)/AMPKα的影响。考察AMPK抑制剂compound C对经6μmol/L澳洲茄边碱作用24 h后细胞中AMPKα、Bcl-2蛋白表达量的影响。结果:与空白组比较,1~12μmol/L澳洲茄边碱作用24、48 h均可显著降低细胞存活率(P<0.05),且有浓度依赖趋势,其半数抑制浓度分别为8.310、7.996μmol/L;经6μmol/L澳洲茄边碱作用10 d后细胞的克隆形成率显著降低(P<0.05)。6、8μmol/L澳洲茄边碱作用后细胞凋亡率和细胞中Bax、caspase-3 mRNA及cleaved caspase-3蛋白(除8μmol/L外)表达量、p-AMPKα/AMPKα(除8μmol/L外)均显著升高(P<0.05),Bcl-2 mRNA及蛋白表达量均显著降低(P<0.05),部分指标变化有浓度依赖性。compound C可以抑制AMPKα蛋白表达并逆转澳洲茄边碱对Bcl-2蛋白的抑制作用。结论:澳洲茄边碱可以抑制HepG2细胞增殖,诱导其凋亡,作用机制可能与激活AMPK信号通路有关。OBJECTIVE:To explore the effects of solamargine on the growth and apoptosis of human hepatocarcinoma cells HepG2 and its underlying mechanism.METHODS:The effects of 0(blank group)-12μmol/L solamargine treatment of 24,48 h on survival rate of HepG2 cells were investigated.The effects of 0(blank group),6μmol/L solamargine treatment of 10 days on cell clone formation were also investigated.The effects of 0(blank group),4,6,8μmol/L solamargine for 24 h on the apoptotic rate of cells,mRNA expression of Bcl-2,Bax and caspase-3,protein expression of Bcl-2 and cleaved caspase-3 as well as ratio of p-AMPKαto AMPKαwere all tested.The effects of AMPK inhibitor as compound C on the protein expression of AMPKαand Bcl-2 in cells were investigated after treated with 6μmol/L solamargine for 24 h.RESULTS:Compared with blank group,1-12μmol/L solamargine for 24,48 h could significantly decrease the survival rates of cells(P<0.05)in a concentration-dependent manner;IC50 of them were 8.310 and 7.996μmol/L,respectively;the rate of cell clone formation was decreased significantly after treated with 6μmol/L solamargine for 10 days(P<0.05).The apoptotic rate of HepG2 cells,mRNA expression of Bax and caspase-3,protein expression of cleaved caspase-3(except for 8μmol/L)as well as ratio of p-AMPKαto AMPKα(except for 8μmol/L)were all increased significantly after treated with 6,8μmol/L solamargine(P<0.05);mRNA and protein expression of Bcl-2 were decreased significantly(P<0.05);the changes of some indexes were in a concentration-dependent manner.The compound C could inhibit protein expression of AMPKα,and reverse the inhibitory effect of solamargine on Bcl-2 protein.CONCLUSIONS:Solamargine can inhibit the proliferation of HepG2 cells and induce apoptosis,the mechanism of which may be associated with activating AMPK signaling pathway.

关 键 词：澳洲茄边碱 人肝癌HEPG2细胞 生长 凋亡 磷酸化腺苷一磷酸活化蛋白激酶信号通路 

分 类 号：R735.7[医药卫生—肿瘤] R965[医药卫生—临床医学]