鳖甲煎丸调控AKT/mTOR信号通路在肝癌细胞有氧糖酵解中的作用  

Effect of Biejia Decoction Pill on aerobic glycolysis in hepatocellular carcinoma by regulating the protein kinase B/mammalian target of rapamycin signaling pathway

作　　者：谭钦文 黄晶晶[2,3] 钟瑞熙 杜沅沁 徐健 农金丽 彭玉姣 TAN Qinwen;HUANG Jingjing;ZHONG Ruixi;DU Yuanqin;XU Jian;NONG Jinli;PENG Yujiao(Graduate School of Guangxi University of Chinese Medicine,Nanning 530023,China;Department of Spleen,Stomach and Hepatology,The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,China;Guangxi Key Laboratory of Translational Medicine for Treating High-Incidence Infectious Diseases with Integrative Medicine,Nanning 530023,China)

机构地区：[1]广西中医药大学研究生院,南宁530023 [2]广西中医药大学第一附属医院脾胃肝病科,南宁530023 [3]广西高发传染病中西医结合转化医学重点实验室,南宁530023

出　　处：《临床肝胆病杂志》2025年第2期300-306,共7页Journal of Clinical Hepatology

基　　金：广西自然科学基金(2022GXNSFAA035460,2024GXNSFDA010005);广西研究生教育创新计划项目(YCSW2023395,YCSY2023021);广西中医药大学引进博士科研启动基金项目(2022BS026)。

摘　　要：目的利用细胞实验探索鳖甲煎丸对肝细胞癌增殖、迁移及有氧糖酵解的抑制作用,并探索其内在机制。方法选取人肝癌细胞株(Huh7)作为研究对象,随机将SD大鼠分为空白血清组、鳖甲煎丸高、中、低剂量组及抑制剂组,制备含药大鼠血清,用以孵育Huh7细胞。CCK-8、划痕实验探索鳖甲煎丸对肝癌细胞的增殖、迁移的影响,糖酵解限速酶及代谢产物检测探索鳖甲煎丸对肝癌细胞有氧糖酵解的影响,RT-qPCR、Western Blot实验探索鳖甲煎丸对AKT/mTOR信号通路的mRNA、相关蛋白及磷酸化的表达。多组间比较采用单因素方差分析,进一步两两比较采用LSD-t或Dunnettg T3检验。结果与空白血清组相比,鳖甲煎丸组的OD值、不同时间段的迁移率、糖酵解限速酶(己糖激酶、磷酸果糖激酶、丙酮酸激酶)及糖酵解代谢产物(丙酮酸、乳酸、ATP)检测均显著降低(P值均<0.05);RT-qPCR结果显示,与空白血清组比较,鳖甲煎丸高、中、低剂量组的mTOR mRNA表达水平均下调,高、低剂量组的AKT mRNA表达水平下调(P值均<0.05);Western Blot结果显示,与空白血清组比较,鳖甲煎丸高、中、低剂量组的mTOR相关蛋白及磷酸化蛋白表达水平均下调,高、中剂量组的AKT相关蛋白及磷酸化蛋白表达水平均下调(P值均<0.05)。结论初步验证了鳖甲煎丸含药血清可以抑制人肝癌细胞Huh7细胞的有氧糖酵解,从而抑制其增殖、迁移,其机制可能与抑制AKT/mTOR信号通路的相关蛋白表达有关。Objective To investigate the inhibitory effect of Biejia Decoction Pill on the proliferation,migration,and aerobic glycolysis of hepatocellular carcinoma(HCC)using cell experiments,as well as related mechanisms.Methods Human liver cancer cell line Huh7 was selected,and Sprague-Dawley rats were randomly divided into blank serum group,inhibitor group,and high-,middle-,and low-dose Biejia Decoction Pill groups.Rat serum containing the drug was prepared for the incubation of Huh7 cells.CCK8 assay and scratch assay were used to explore the effect of Biejia Decoction Pill on the proliferation and migration of HCC cells;glycolytic rate-limiting enzymes and metabolites were measured to explore the effect of Biejia Decoction Pill on aerobic glycolysis of liver cancer cells;RT-qPCR and Western blot were used to explore the effect of Biejia Decoction Pill on the mRNA expression,related proteins,and phosphorylation of the protein kinase B(AKT)/mammalian target of rapamycin(mTOR)signaling pathway.A one-way analysis of variance was used for comparison between multiple groups,and the least significant difference t-test or the Dunnett’s T3 test were used for further comparison between two groups.Results Compared with the blank serum group,the Biejia Decoction Pill groups had significant reductions in OD value,migration rate during different periods of time,glycolytic rate-limiting enzymes(hexokinase,phosphofructokinase,pyruvate kinase),and glycolytic metabolites(pyruvate,lactic acid,ATP)(all P<0.05).RT-qPCR results showed that compared with the blank serum group,the high-,middle-,and low-dose Biejia Decoction Pill groups had a significant reduction in the mRNA expression level of mTOR,and the high-and low-dose Biejia Decoction Pill groups had a significant reduction in the mRNA expression level of AKT(all P<0.05).Western blot results showed that compared with the blank serum group,the high-,middle-,and low-dose Biejia Decoction Pill groups had significant reductions in the expression levels of mTOR-related proteins and phospho

关 键 词：鳖甲煎丸 肝肿瘤 实验性 瓦尔堡效应 肿瘤学 

分 类 号：R285.5[医药卫生—中药学]