基于AMPK/mTOR信号通路探讨人参-黄芪对人胃癌细胞自噬的影响  

Exploring the effect of ginseng and astragalus on autophagy of human gastric cancer cells based on AMPK/mTOR signaling pathway

作　　者：张海洋 陈思鼎 施妙璇 郑薇 韩美奕 丁治国[1] 季双双 田明[1] ZHANG Haiyang;CHEN Siding;SHI Miaoxuan;ZHENG Wei;HAN Meiyi;DING Zhiguo;JI Shuangshuang;TIAN Ming(Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China)

机构地区：[1]北京中医药大学东直门医院,北京100700 [2]密歇根大学安娜堡分校胃肠病学与肝病学部,美国密歇根州MI 48105

出　　处：《陕西中医》2024年第6期723-727,共5页Shaanxi Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(81673972);北京中医药大学青年教师科研项目(2018-JYBZZ-JS081)。

摘　　要：目的:探究益气扶正药人参-黄芪对人胃癌细胞自噬的影响,并探讨腺苷酸活化蛋白激酶(AMPK)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路在其中的作用机制。方法:将人胃癌细胞株(SGC-7901)随机分为对照组(人参-黄芪0 mg/ml)和人参-黄芪组(1、2、4、8、16、32、64 mg/ml),经药物处理后,CCK-8法观察人胃癌细胞增殖情况;筛选最佳浓度并将其作为后续实验中的实验组;MDC免疫荧光染色观察细胞自噬情况;Western blot法检测人胃癌细胞自噬相关蛋白(Beclin-1、LC3B)及AMPK/mTOR信号通路相关蛋白(AMPK、p-AMPK、mTOR及p-mTOR)的表达情况。结果:CCK-8法结果显示,人参-黄芪组人胃癌细胞增殖作用降低(P<0.01),且呈浓度依赖性;MDC染色荧光检测显示实验组的荧光强度显著增强(P<0.01),表明人参-黄芪可以诱导人胃癌细胞产生自噬;Western blot结果显示,实验组的Beclin-1、LC3B、p-AMPK/AMPK蛋白表达水平上升(P<0.01),p-mTOR/mTOR蛋白表达水平下降(P<0.01)。结论:人参-黄芪配伍能够抑制人胃癌细胞的增殖、促进人胃癌细胞自噬,其机制可能与激活AMPK/mTOR信号通路相关。Objective:To investigate the effect of ginseng and astragalus on autophagy of human gastric cancer cells,and to explore the mechanism of adenosine monophosphate-activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR)signaling pathway in this process.Methods:The human gastric cancer cell lines(SGC-7901)were randomly divided into control group(ginseng and astragalus 0 mg/ml)and ginseng and astragalus groups(1,2,4,8,16,32,64 mg/ml).After medicine treatment,the proliferation of human gastric cancer cells was observed by CCK-8 method.The optimal concentration was screened and used as the experimental group in the subsequent experiments.The autophagy of the cells was observed by MDC immunofluorescence staining.The expressions of autophagy-related proteins(Beclin-1,LC3B)and AMPK/mTOR signaling pathway-related proteins(AMPK,p-AMPK,mTOR and p-mTOR)in human gastric cancer cells were detected by Western blot.Results:The results of CCK-8 showed that the proliferation of human gastric cancer cells in ginseng and astragalus group was decreased(P<0.01),and the proliferation was concentration-dependent.MDC fluorescence detection showed that the fluorescence intensity of the experimental group was significantly enhanced(P<0.01),indicating that ginseng and astragalus could induce autophagy in human gastric cancer cells.Western blot results showed that the protein expression levels of Beclin-1,LC3B and p-AMPK/AMPK in the experimental group increased(P<0.01),while the protein expression levels of p-mTOR/mTOR decreased(P<0.01).Conclusion:The combination of ginseng and Astragalus can inhibit the proliferation and promote the autophagy of human gastric cancer cells.The activation of the AMPK/mTOR signaling pathway may underlie this mechanism.

关 键 词：胃癌 人参 黄芪 腺苷酸激活蛋白激酶 雷帕霉素靶蛋白 自噬 

分 类 号：R735.2[医药卫生—肿瘤]