归芪益元膏联合顺铂对Lewis肺癌小鼠SIRT3/AMPK/mTOR/HIF-1α信号通路的影响  

Effects of Guiqi Yiyuan Ointment combined with Cisplatin on SIRT3/AMPK/mTOR/HIF-1a signaling pathway in Lewis lung cancer mouse

作　　者：梁建庆 杨玉萍 李金田[2,3] 李娟 LIANG Jianqing;YANG Yuping;LI Jintian;LI Juan(School of Basic Medicine,Gansu University of Chinese Medicine,Lanzhou 730000,China;The Key Laboratory of Ministry of Education of Dunhuang Medicine and Transformation,Lanzhou 730000,China;Clinical College of Traditional Chinese Medicine,Gansu University of Chinese Medicine,Lanzhou 730000,China)

机构地区：[1]甘肃中医药大学基础医学院,兰州730000 [2]敦煌医学与转化教育部重点实验室,兰州730000 [3]甘肃中医药大学中医临床学院,兰州730000

出　　处：《中华中医药杂志》2024年第8期3958-3963,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.82160872);甘肃省中医药研究中心开放课题(No.zyzx-2020-zx20)。

摘　　要：目的:基于SIRT3/AMPK/mTOR/HIF-1α信号通路探讨归芪益元膏联合顺铂对Lewis肺癌小鼠的干预效应及机制。方法:C57BL/6J小鼠复制Lewis肺癌小鼠模型。将造模小鼠随机分为模型组、顺铂组、中药组及归芪益元膏低、中、高剂量联合顺铂组,每组10只,药物干预13 d并记录其体质量。末次给药24 h后处死小鼠;苏木精-伊红(HE)染色观察小鼠肿瘤组织病理学变化;ELISA检测各治疗组小鼠血清中TGF-β3、EGF含量;免疫荧光法检测各治疗组小鼠肿瘤组织中Ki67蛋白表达情况;实时荧光定量PCR和Western Blot检测肿瘤组织SIRT3/AMPK/mTOR/HIF-1α通路相关蛋白和基因的表达。结果:模型组小鼠体质量显著下降(P<0.05);经治疗后,归芪益元膏各剂量联合顺铂组小鼠体质量显著升高(P<0.05);血清中TGF-β3、EGF含量,肿瘤组织中Ki67、p-mTOR、HIF-1α蛋白及mRNA表达显著下降(P<0.05),肿瘤组织中SIRT3、AMPK蛋白及mRNA表达显著升高(P<0.05),其中归芪益元膏高剂量联合顺铂治疗效果显著。结论:归芪益元膏联合顺铂能够通过靶向调控SIRT3/AMPK/mTOR/HIF-1α信号通路相关蛋白的表达,抑制其肿瘤的生长。Objective:To investigate the intervention effects and mechanism of Guiqi Yiyuan Ointment combined with Cisplatin in Lewis lung cancer mice based on SIRT3/AMPK/mTOR/HIF-1αsignaling pathway.Methods:The Lewis lung cancer mouse model was replicated with SPF grade C57BL/6J mice.The model mice were randomly divided into model group,Cisplatin group,Chinese medicine group and Guiqi Yiyuan Ointment low,medium and high dose combined with Cisplatin group,10 mice in each group,and their body weight was recorded for 13 d of drug intervention.After 24 h of final administration,the tumor inhibition rate of each drug group was measured;HE was used to observe the histopathological changes of tumors in mice;ELISA kits were used to detect the changes of TGF-β3 and EGF in the serum of each treatment group:;immunofluorescence method was used to detect the expression of Ki67 protein in the tumors of mice in each treatment group,the expression of SIRT3/AMPK/mTOR/HIF-1αpathway related proteins and genes in the tumor tissues was detected by real-time fluorescence quantitative polymerase chain reaction(RT-PCR)and Western Blot.Results:In the model group,the average body weight of mice decreased significantly(P<0.05).After treatment,the body weight of mice in Guiqi Yiyuan Ointment low,medium and high dose combined with Cisplatin group increased gradually(P<0.05),the content of TGF-β3 and EGF in serum and the expression of Ki67,p-mTOR and HIF-1αprotein/mRNA in tumor tissue decreased gradually(P<0.05),the expression of SIRT3 and AMPK protein/mRNA increased gradually in tumor tissues(P<0.05),and the effects of Guiqi Yiyuan Ointment high dose combined with Cisplatin was significant.Conclusion:Guiqi Yiyuan Ointment combined with Cisplatin can inhibit the growth of tumor by targeting the expression of SIRT3/AMPK/mTOR/HIF-1αsignaling pathway-related proteins.

关 键 词：归芪益元膏 顺铂 肺癌 沉默调节蛋白3/腺苷酸激活蛋白激酶/哺乳动物雷帕霉素靶蛋白/缺氧诱导因子1α 

分 类 号：R285.5[医药卫生—中药学]