和胃降逆胶囊通过PI3K/AKT信号通路调控人胃癌AGS细胞增殖与凋亡机制研究  

Mechanism of Hewei Jiangni Capsule(和胃降逆胶囊)Regulating Proliferation and Apoptosis of Human Gastric Cancer AGS Cells Through PI3K/AKT Signaling Pathway

作　　者：窦建卫[1] 杨艺[1,2] 杨小蒨 杨燕莹 李高彪 唐锐 朱宇红 DOU Jianwei;YANG Yi;YANG Xiaoqian;YANG Yanying;LI Gaobiao;TANG Rui;ZHU Yuhong(Xi’an Jiaotong University,Xi’an 710061,China;Baoji Central Hospital,Baoji 721008,China;College of Basic Medical Sciences,Gansu University of Traditional Chinese Medicine,Lanzhou 730000,China;Xiyuan Clinical College of Beijing University of Chinese Medicine,Beijing 100029,China;Xiyuan Hospital of China Academy of Chinese Medical Science,Beijing 100091,China;Central Hospital of Xianyang,Xianyang 712099,China)

机构地区：[1]西安交通大学,陕西西安710061 [2]宝鸡市中心医院,陕西宝鸡721008 [3]甘肃中医药大学基础医学院,甘肃兰州,730000 [4]北京中医药大学西苑临床医学院,北京100029 [5]中国中医科学院西苑医院,北京100091 [6]咸阳市中心医院,陕西咸阳712099

出　　处：《山东中医药大学学报》2022年第1期101-107,共7页Journal of Shandong University of Traditional Chinese Medicine

基　　金：陕西省重点研发计划项目(编号:2019SF-013)。

摘　　要：目的:探究和胃降逆胶囊对人胃癌AGS细胞增殖和凋亡的影响及其作用机制。方法:用不同药物剂量(3 g/kg、6 g/kg、12 g/kg)制备的和胃降逆胶囊含药血清分别干预人胃癌AGS细胞,分为和胃降逆胶囊含药血清低剂量组、中剂量组、高剂量组及空白组,分别干预AGS细胞24 h、48 h、72 h后,采用四甲基偶氮唑蓝(MTT)法检测细胞增殖情况,采用流式细胞仪检测细胞周期分布情况,蛋白质印迹法(Western blot)检测细胞相关蛋白磷酸化磷脂酰肌醇3-激酶(p-PI3K)、磷酸化蛋白激酶B(p-AKT)、促凋亡基因(Bax)、抑细胞凋亡蛋白(Bcl-2)、细胞周期蛋白D1(Cyclin D1)和细胞周期蛋白依赖性抑制剂(p21)的表达情况;实时荧光定量聚合酶链式反应(RT-PCR)法检测细胞凋亡相关基因Bax、Bcl-2、Cyclin D1和p21 mRNA表达情况。结果:随着和胃降逆胶囊含药血清作用时间的延长,各药物组细胞活力逐渐降低,同剂量组24 h、48 h、72 h比较差异有统计学意义(P<0.05),48 h与72 h比较差异无统计学意义。在同一干预时间点,细胞活力随着药物血清浓度的增加而降低。各组含药血清处理AGS细胞48 h后,G0/G1期细胞比值与空白组比较,差异具有统计学意义(P<0.05);和胃降逆胶囊可以下调AGS细胞中p-PI3K、p-AKT蛋白及抑制凋亡蛋白Bcl-2和Cyclin D1的表达(P<0.05),上调AGS细胞中促凋亡蛋白Bax和p21的表达(P<0.05);和胃降逆胶囊可以下调AGS细胞中抑制凋亡基因Bcl-2和Cyclin D1 mRNA的表达(P<0.05),上调AGS细胞中促凋亡基因Bax和p21 mRNA的表达(P<0.05),且呈现出浓度相关性。结论:和胃降逆胶囊通过抑制PI3K/AKT通路的活化,调控下游周期阻滞相关基因和凋亡相关基因的表达,进而抑制人胃癌AGS细胞增殖并促进AGS细胞凋亡。Objective:To investigate the effect of Hewei Jiangni Capsule(和胃降逆胶囊)on proliferation and apoptosis of human gastric cancer AGS cells,and explore its mechanism.Methods:Human gastric cancer AGS cells were intervened with different drug doses(3 g/kg,6 g/kg,12 g/kg)and divided into Hewei Jiangni Capsule containing serum low-dose group,medium-dose group,high-dose group and blank group.The cells were intervened for 24 h,48 h and 72 h respectively.Methyl thiazolyl tetrazolium(MTT)method was used to detect the proliferation of AGS cells.Flow cytometry was used to detect the cell cycle distribution.Western blot was used to detect the expression of phosphorylation phosphatidylinositol 3-hydroxy kinase(p-PI3K),phosphorylated protein kinase B(p-AKT),pro-apoptotic gene(Bax),anti-apoptotic protein(Bcl-2),Cyclin D1 and cyclin dependent inhibitors(p21).The expression of apoptosis related genes Bax,Bcl-2,cyclin D1 and p21 mRNA was detected by real-time fluorescence quantitative polymerase chain reaction(RT-PCR).Results:With the prolongation of time by Hewei Jiangni Capsule medicated serum,the cell viability of each drug group decreased gradually,and there was statistically significant at 24 h and 48 h and 72 h at the same dose(P<0.05),but there was no statistical significance between 48 h and 72 h.At the same intervention time point,the cell viability decreased with the increase of drug serum concentration.Compared with the blank group,the ratio of G0/G1 phase cells was statistical significance(P<0.05).Hewei Jiangni Capsule could down-regulate the expression of p-PI3K,p-AKT protein and inhibit the expression of apoptosis protein Bcl-2 and Cyclin D1 in AGS cells(P<0.05),and up-regulate the expression of apoptotic proteins Bax and p21 in AGS cells(P<0.05).Hewei Jiangni Capsule could down-regulate the mRNA expression of apoptosis inhibiting gene Bcl-2 and Cyclin D1 in AGS cells(P<0.05),and up-regulate the mRNA expression of proapoptotic gene Bax and p21 in AGS cells(P<0.05),and presented a concentration correlation.Concl

关 键 词：和胃降逆胶囊 人胃癌AGS细胞 细胞增殖 细胞凋亡 磷脂酰肌醇3-激酶/蛋白激酶B通路 四甲基偶氮唑蓝法 蛋白质印迹法 实时荧光定量聚合酶链式反应 

分 类 号：R285.5[医药卫生—中药学]