芪竭消痞汤含药血清对MNNG诱导的人胃黏膜上皮细胞自噬及凋亡的影响OA
Effects of the Drug-Containing Serum of Qijie Xiaopi Decoction on Autophagy and Apoptosis of Human Gastric Mucosal Epithelial Cells Induced by MNNG
目的:观察芪竭消痞汤对MNNG诱导的胃黏膜上皮细胞恶性转化的影响,并基于PI3K/AKT/mTOR信号通路探讨其治疗胃癌前病变的作用机制.方法:CCK-8法检测MNNG诱导人胃黏膜上皮细胞GES-1 恶性转化的有效浓度,建立体外PLGC细胞模型(MC细胞).设空白对照组、模型组和芪竭消痞汤含药血清高、中、低浓度组和Ly294002(PI3K抑制剂)组.CCK-8法检测芪竭消痞汤对MC细胞增殖的影响,MDC染色观察MC细胞自噬情况,流式细胞术测定细胞凋亡率,qPCR法检测Bcl-2、Bax mRNA表达,蛋白免疫印迹法检测 PI3K、p-AKT、p-mTOR、AKT、p-mTOR、p62、LC3、Beclin-1蛋白表达.结果:采用浓度为 20 μmol/L的MNNG干预GES-1 细胞 48 h后,细胞抑制率达到 50%左右,且在此浓度下GES-1 细胞体现出恶性转化的生物学特性;同时间点与GES-1 细胞组比较,MC细胞活力显著升高(P<0.01).与模型组比较,芪竭消痞汤含药血清中、高浓度组和Ly294002 组细胞活力显著降低,自噬体数量、凋亡率、Bax mRNA表达和LC3、Beclin-1蛋白表达显著升高,Bcl-2 mRNA和p62、PI3K、p-AKT、p-mTOR蛋白表达显著降低(P<0.05 或P<0.01).结论:芪竭消痞汤能通过抑制MC细胞的增殖,促进细胞自噬及凋亡,从而发挥治疗胃癌前病变的作用,相关机制可能与抑制PI3K/AKT/mTOR信号通路有关.
Objective:To observe the effect of Qijie Xiaopi Decoction on the malignant transformation of gastric mucosal epithelial cells induced by MNNG,and to explore its mechanism of treating precancerous lesions of gastric cancer based on the PI3K/AKT/mTOR signaling pathway.Methods:The CCK-8 assay was used to determine the effective concentration of MNNG for inducing the malignant transformation of human gastric mucosal epithelial cells GES-1,and in vitro PLGC cell model(MC cells)was established.The blank control group,model group,the high,medium and low concentration groups of Qijie Xiaopi Decoction drug-containing serum,Ly294002(PI3K inhibitor)group were set up.The effect of Qijie Xiaopi Decoction on the proliferation of MC cells was detected by CCK-8 meth-od.The autophagy of MC cells was observed by MDC staining.The apoptosis rate of cells was determined by flow cytometry.The mRNA expressions of Bcl-2 and Bax were detected by qPCR.The protein expressions of PI3K,p-AKT,p-mTOR,AKT,p-mTOR,p62,LC3 and Beclin-1 were detected by Western blotting.Results:After interfering with GES-1 cells with MNNG at a concentration of 20 μmol/L for 48 hours,the cell inhibition rate reached about 50%,and at this concentration,GES-1 cells exhibited the biological characteristics of malignant transformation.At the same time points,compared with the GES-1 cell group,the viability of MC cells was significantly in-creased(P<0.01).Compared with the model group,the cell viability in the medium and high concentration groups of drug-containing serum of Qijie Xiaopi Decoction and the Ly294002 group was significantly decreased,while the number of autophagosomes,apoptosis rate,expression of Bax mRNA,and protein expressions of LC3 and Beclin-1 were significantly increased.The expressions of Bcl-2 mR-NA and p62,PI3K,p-AKT,p-mTOR proteins were significantly decreased(P<0.05 or P<0.01).Conclusion:Qijie Xiaopi Decoction can exert therapeutic effects on precancerous lesions of gastric cancer by inhibiting the proliferation of MC cells,promoting autophagy and apoptosis of cells.The related mechanism may be associated with the inhibition of the PI3K/AKT/mTOR signaling pathway.
单意;白敏;马洋;陈涵斐;史明珠;虎峻瑞
宁夏医科大学中医学院,宁夏 银川 750004||宁夏医科大学少数民族医药现代化教育部重点实验室,宁夏 银川 750004宁夏医科大学中医学院,宁夏 银川 750004||宁夏医科大学少数民族医药现代化教育部重点实验室,宁夏 银川 750004宁夏医科大学中医学院,宁夏 银川 750004||宁夏医科大学少数民族医药现代化教育部重点实验室,宁夏 银川 750004宁夏医科大学中医学院,宁夏 银川 750004||宁夏医科大学少数民族医药现代化教育部重点实验室,宁夏 银川 750004宁夏医科大学中医学院,宁夏 银川 750004||宁夏医科大学少数民族医药现代化教育部重点实验室,宁夏 银川 750004宁夏医科大学中医学院,宁夏 银川 750004||宁夏医科大学少数民族医药现代化教育部重点实验室,宁夏 银川 750004
医药卫生
芪竭消痞汤胃癌前病变MC细胞PI3K/AKT/mTOR自噬凋亡
Qijie Xiaopi DecoctionPrecancerous lesions of gastric cancerMC cellsPI3K/AKT/mTORAutophagyApoptosis
《中药材》 2025 (11)
2860-2867,8
宁夏回族自治区重点研发项目(2022BEG03150)宁夏医科大学科学研究资助项目研究计划(XZ2022006)
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