小檗碱基于AMPK/mTOR通路对EC9706细胞能量代谢的影响  

Effect of Berberine on Energy Metabolism in EC9706 Cells Based on AMPK/mTOR Pathway

作　　者：刘洋[1] 陈星[1] 周哲旭 刘娅茹 胡啸博 陈玉龙[1] LIU Yang;CHEN Xing;ZHOU Zhexu;LIU Yaru;HU Xiaobo;CHEN Yulong(Henan Key Laboratory of Signal Transduction,Henan University of Chinese Medicine,Zhengzhou 450046,Henan,China)

机构地区：[1]河南中医药大学河南省方证信号传导重点实验室,河南郑州450046

出　　处：《中华中医药学刊》2025年第1期97-101,I0012,I0013,共7页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81873285,82074313,82274438)。

摘　　要：目的通过观察小檗碱对EC9706细胞能量代谢及相关通路的影响从而探究其机制。方法常规培养细胞,MTT法筛药,选择15、30μg/mL两个质量浓度,通过流式细胞术检测细胞周期、凋亡水平,能量代谢分析系统检测细胞糖酵解潜能及线粒体呼吸能力,蛋白质印迹实验探究细胞中p-AMPK/AMPK及下游p-mTOR/mTOR蛋白表达水平变化,通过荧光实时定量PCR(quantitative real-time PCR,qPCR)在基因水平上验证AMPK及下游mTOR的表达水平。结果小檗碱能够明显抑制EC9706细胞增殖(P<0.01),促进EC9706细胞凋亡(P<0.05),与对照组相比,小檗碱加药组G0/G1期明显降低(P<0.05),S期占比明显升高(P<0.01)。小檗碱能够抑制EC9706细胞的糖酵解潜能及线粒体呼吸能力(P<0.01)且具有浓度依赖性。小檗碱能够促进p-AMPK/AMPK(P<0.01)并抑制p-mTOR/mTOR蛋白表达(P<0.05)。在基因水平上,小檗碱促进了AMPK表达(P<0.05),同时促进了mTOR的表达(P<0.01)。结论小檗碱可以通过能量代谢途径抑制食管癌EC9706细胞的增殖,促进食管癌细胞凋亡并阻碍其分裂。Objective To investigate the mechanism of berberine on energy metabolism and related pathways in EC9706 cells.Method The cells were routinely cultured,MTT screened,and two mass concentrations of 15μg/mL and 30μg/mL were selected.The cell cycle and apoptosis were detected by flow cytometry,the glycolytic potential and mitochondrial respiratory capacity were detected by energy metabolism analysis system,and the changes in the expression of phosphorylated adenylate activated protein kinase(p-AMPK)/adenylate activated protein kinase(AMPK)and the downstream phosphorylated mammalian target of rapamycin(p-mTOR)/mammalian target of rapamycin(mTOR)proteins were explored by Western Blotting.The expression levels of AMPK and downstream mTOR were verified at the gene level by quantitative real-time PCR(qPCR).Results Berberine significantly inhibited the proliferation of EC9706 cells(P<0.01)and promoted the apoptosis of EC9706 cells(P<0.05).Compared with the control group,berberine and drug group showed a significant decrease in the G0/G1 phase(P<0.05),and a significant increase in the proportion of S phase(P<0.01).Berberine inhibited the glycolytic potentia and mitochondrial respiration of EC9706 cells(P<0.01)in a concentration-dependent manner,and berberine promoted p-AMPK/AMPK(P<0.01)and inhibited the expression of p-mTOR/mTOR protein(P<0.05).At the gene level,berberine promoted AMPK expression(P<0.05)as well as mTOR expression(P<0.01).Conclusion Berberine can inhibit the proliferation of oesophageal cancer EC9706 cells through energy metabolic pathway,promote apoptosis of oesophageal cancer cells and hinder their division.

关 键 词：小檗碱 能量代谢 AMPK/mTOR通路 增殖 周期 凋亡 

分 类 号：R282.71[医药卫生—中药学]