齐墩果酸诱导肝癌细胞凋亡的实验研究  

Experimental Study of Oleanolic Acid Inducing Apoptosis of Hepatic Carcinoma Cells

作　　者：孙阳[1] 孙悦 吴勃岩[1] 陶雪莲 姜德友[1] SUN Yang;SUN Yue;WU Boyan;TAO Xuelian;JIANG Deyou(Heilongjiang University of Chinese Medicine,Harbin 150040,China)

机构地区：[1]黑龙江中医药大学,黑龙江哈尔滨150040

出　　处：《中医药学报》2021年第11期33-37,共5页Acta Chinese Medicine and Pharmacology

基　　金：国家自然科学基金青年基金(81704054);配套课题编号(2017PT08);中国博士后科学基金资助项目(2014M551288);黑龙江省博士后资助项目(LBH-Z13205)。

摘　　要：目的:研究齐墩果酸对人肝癌SMMC-7721细胞的抑制作用,并探讨其机制。方法:体外培养肝癌细胞,制备齐墩果酸自微乳,采用MTT法对细胞存活率进行检测,Elisa法检测细胞培养液中的IL-6含量,AnnexinV-FITC双标法荧光显微镜观察凋亡细胞,流式细胞术检测各组凋亡率,实时荧光定量PCR(quantitative Real-Time PCR)法检测各组细胞Bax、Bcl-2和Akt mRNA表达,蛋白免疫印迹法(Western blot)检测Bax、Bcl-2、Akt和p-Akt蛋白表达。结果:与模型对照组相比,随着齐墩果酸剂量增加,细胞存活率下降(P<0.05),80μg·mL^(-1)齐墩果酸和15μg·mL^(-1)顺铂作用肝癌细胞24 h,细胞培养液中IL-6含量降低(P<0.05),凋亡细胞数量明显高于模型对照组;流式细胞术检测发现,齐墩果酸和顺铂组早期凋亡、晚期凋亡和总凋亡率均高于模型对照组(P<0.05);用药组细胞的Bax mRNA和蛋白表达高于模型对照组(P<0.05),Bcl-2和Akt mRNA表达低于模型对照组(P<0.05),Bcl-2、Akt和p-Akt蛋白表达低于模型对照组(P<0.05)。结论:齐墩果酸通过上调Bax、下调Bcl-2基因表达,抑制Akt信号通路,抑制IL-6产生,诱导细胞凋亡,起到抗肿瘤的作用。Objective:To study the effect of Oleanolic Acid(OA)inhibiting hepatic carcinoma cell SMMC-7721,and to explore its action mechanism.Methods:Cancer cells were cultured in vitro.OA self-microemulsion was prepared.The cell survival rate was measured by MTT method.The content of IL-6 in cell culture medium was detected by Elisa method.The cell apoptosis was observed by AnnexinV-FITC double standard method and fluorescence microscope.The rate of apoptosis was measured by flow cytometry.The mRNA expressions of Bax,Bcl-2 and Akt were detected by qRT-PCR.The protein expressions of Bax,Bcl-2,Akt,and p-Akt were measured by Western blot.Results:Compared to those in the model group,the cell survival rate showed a significant dose-dependent decrease(P<0.05);the content of IL-6 significantly decreased in the culture medium after 24 h interventions of 80μg·mL^(-1)OA and 15μg·mL^(-1) Cisplatin.Flow cytometry results showed that the rates of early apoptosis,late apoptosis and total apoptosis were higher in the OA group and the Cisplatin group than those in the model group.Compared to those in the model group,the mRNA expression and protein expression of Bax were significantly higher(P<0.05),the mRNA expressions of Bcl-2 and Akt were significantly lower(P<0.05),and the protein expressions of Bcl-2,Akt and p-Akt were significantly lower in the OA group and the Ciplatin group(P<0.05).Conclusion:OA can play the anti-tumor role by up-regulating Bax gene expression and down-regulating Bcl-2 gene expression,as well as inhibiting Akt signaling pathway and decreasing IL-6 to induce cell apoptosis.

关 键 词：齐墩果酸 人肝癌SMMC-7721细胞 BAX BCL-2 AKT 

分 类 号：R285.5[医药卫生—中药学]