基于mTOR/HIF-1α/VEGF信号通路探讨抗纤抑癌方对肝癌前病变大鼠模型的调控作用  

Regulatory effect of Kangxian Yiai Prescription in a rat model of precancerous lesions of liver cancer:A study based on the mTOR/HIF-1α/VEGF signaling pathway

作　　者：李志国 马浔 叶永安[3] 杨先照[3] LI Zhiguo;MA Xun;YE Yongan;YANG Xianzhao(Department of Gastroenterology,Fengtai Hospital of Integrated Traditional Chinese and Western Medicine,Beijing 100072,China;Department of Traditional Chinese Medicine,Beijing Xing Yuan Jin Fang Clinic of Chinese Medicine,Beijing 101300,China;Institute of Liver Diseases,Beijing University of Chinese Medicine,Beijing 100700,China)

机构地区：[1]北京市丰台中西医结合医院消化科,北京100072 [2]北京杏园金方国医医院中医科,北京101300 [3]北京中医药大学肝病研究院,北京100700

出　　处：《临床肝胆病杂志》2024年第10期2049-2054,共6页Journal of Clinical Hepatology

基　　金：国家自然科学基金青年科学基金项目(81603555);北京市自然科学基金青年科学基金项目(7174319);北京市丰台中西医结合医院院自然基金(YS2022-01)。

摘　　要：目的研究抗纤抑癌方对肝癌前病变大鼠模型mTOR/HIF-1α/VEGF信号通路的影响。方法40只雄性Wistar大鼠,分为正常组、模型组、抗纤抑癌组和鳖甲软肝组,每组各10只。正常组大鼠腹腔注射生理盐水,剂量为0.4 mL/100 g,其他3组大鼠以50 mg/kg剂量腹腔注射二乙基亚硝胺,制备肝癌前病变大鼠模型。通过免疫组化和Western Blot法检测GST-Pi的表达,实时荧光定量PCR和Western Blot法检测mTOR、HIF-1α、VEGF、M2型丙酮酸激酶(PKM2)、葡萄糖载体蛋白1(GLUT-1)mRNA及蛋白的表达。计量资料多组间比较采用单因素方差分析或Kruskal-Wallis H秩和检验,进一步两两比较采用LSD-t检验。结果与正常组比较,模型组大鼠肝组织GST-Pi蛋白表达显著升高(P<0.01);与模型组比较,抗纤抑癌组GST-Pi蛋白表达水平显著降低(P<0.05)。与正常组比较,模型组大鼠肝组织GLUT1及PKM2 mRNA的表达均显著升高(P值均<0.01);与模型组比较,鳖甲软肝组及抗纤抑癌组GLUT1 mRNA的表达均显著降低(P值均<0.05)。与正常组比较,模型组大鼠肝组织GLUT1及PKM2的蛋白表达均显著升高(P值均<0.01)。与正常组比较,模型组大鼠肝组织mTOR、HIF-1α及VEGF的mRNA表达均显著升高(P值均<0.01);与模型组比较,鳖甲软肝组mTOR及VEGF的mRNA的表达均显著降低(P值均<0.05),抗纤抑癌组mTOR及VEGF mRNA的表达亦显著降低(P值均<0.01)。与正常组比较,模型组大鼠肝组织mTOR、HIF-1α、VEGF的蛋白表达均显著升高(P值均<0.01);与模型组相比,鳖甲软肝组只有mTOR的蛋白表达显著降低(P<0.01),抗纤抑癌组mTOR、HIF-1α、VEGF的蛋白表达均显著降低(P值均<0.05);与鳖甲软肝组相比,抗纤抑癌组mTOR的蛋白表达较高(P<0.01)。结论抗纤抑癌方可通过调控mTOR/HIF-1α/VEGF信号抑制肝癌前病变。Objective To investigate the effect of Kangxian Yiai Prescription(KXYA)on the mTOR/HIF-1α/VEGF signaling pathway in a rat model of precancerous lesions of liver cancer.Methods A total of 40 male Wistar rats were divided into normal group,model group,KXYA group,and Biejia Rangan Tablets(BJRG)group,with 10 rats in each group.The rats in the normal group were given intraperitoneal injection of normal saline at a dose of 0.4 mL/100 g,and those in the other three groups were given intraperitoneal injection of diethylnitrosamine at a dose of 50 mg/kg to establish a rat model of the precancerous lesions of liver cancer.Immunohistochemistry and Western Blot were used to measure the expression level of GST-Pi,and quantitative real-time PCR and Western Blot were used to measure the mRNA and protein expression levels of mTOR,HIF-1α,VEGF,PKM2,and GLUT1.A oneway analysis of variance or the Kruskal-Wallis H test was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups.Results Compared with the normal group,the model group had a significant increase in the protein expression level of GST-Pi in liver tissue(P<0.01),and compared with the model group,the KXYA group had a significant reduction in the protein expression level of GST-Pi(P<0.05).Compared with the normal group,the model group had significant increases in the mRNA expression levels of GLUT1 and PKM2 in liver tissue(P<0.01),and compared with the model group,the BJRG group and the KXYA group had a significant reduction in the mRNA expression level of GLUT1(P<0.05).Compared with the normal group,the model group had significant increases in the protein expression levels of GLUT1 and PKM2 in liver tissue(P<0.01).Compared with the normal group,the model group had significant increases in the mRNA expression levels of mTOR,HIF-1α,and VEGF in liver tissue(P<0.01);compared with the model group,the BJRG group had significant reductions in the mRNA expression levels of mTOR

关 键 词：肝肿瘤 信号传导 抗纤抑癌方 大鼠 Wistar 

分 类 号：R285.5[医药卫生—中药学]