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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

李志国 马浔 叶永安 杨先照

引用本文:
Citation:

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

DOI: 10.12449/JCH241019
基金项目: 

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)

伦理学声明:本研究方案于2017年5月经由北京中医药大学东直门医院实验动物伦理委员会审批,批号:17-05,符合实验室动物管理与使用准则。
利益冲突声明:本文不存在任何利益冲突。
作者贡献声明:叶永安、杨先照对研究的思路或设计有关键贡献;马浔、李志国参与了研究数据的获取分析解释过程;李志国、杨先照参与起草及修改文章关键内容。
详细信息
    通信作者:

    李志国, lizhiguo1888@163.com (ORCID: 0000-0003-3533-2542)

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

Research funding: 

National Natural Science Foundation of China Youth Science Fund Project (81603555);

Beijing Natural Science Foundation Youth Science Fund Project (7174319);

Beijing Fengtai Integrated Traditional Chinese and Western Medicine Hospital Natural Science Foundation (YS2022-01)

More Information
  • 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。

     

  • 图  1  大鼠肝组织GST-Pi免疫组化(×400)

    注: a,正常组;b,模型组;c,鳖甲软肝组;d,抗纤抑癌组。

    Figure  1.  Rat liver tissue GST-Pi immunohistochemistry (×400)

    图  2  各组大鼠肝组织GST-Pi蛋白表达情况

    Figure  2.  Expression of GST-Pi Protein in rat liver tissues

    图  3  大鼠肝组织GLUT1和PKM2的mRNA表达

    Figure  3.  mRNA expression of GLUT1 and PKM2 in rat liver tissues

    图  4  大鼠肝组织GLUT1和PKM2的蛋白表达

    Figure  4.  Protein expression of GLUT1 and PKM2 in rat liver tissues

    图  5  大鼠肝组织mTOR、HIF-1α、VEGF的 mRNA表达

    Figure  5.  mRNA expression of mTOR, HIF-1α, and VEGF in rat liver tissues

    图  6  大鼠肝组织mTOR、HIF-1α、VEGF蛋白表达

    Figure  6.  Protein expression of mTOR, HIF-1α, and VEGF in rat liver tissues

    表  1  实时荧光定量PCR引物序列

    Table  1.   Real time fluorescence quantitative PCR primer sequence

    引物名称 引物序列(5'-3') 扩增产物长度(bp)
    Rat-mTOR F:TGTCAGCCTGTCAGAATCCA 74
    R:CCATGTTGACCAGCATTTCA
    Rat-HIF-1α F:TGGAAGCACTAGACAAAGCTCA 78
    R:TTGACCATATCGCTGTCCAC
    Rat-VEGF F:GAGTTAAACGAACGTACTTGCAGA 90
    R:TCTAGTTCCCGAAACCCTGA
    Rat-PKM2 F:GGAGAAGTGCGATGAGAACAT 141
    R:TCTGTCACCAGGTAGTCAGCAC
    Rat-GLUT1 F:GTATCCTGTTGCCCTTCTGC 95
    R:TCGAAGCTTTTTCAGCACAC
    GAPDH F:TCATTGACCTCAACTACATGG 131
    R:TCGCTCCTGGAAGATGGTG
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  • 收稿日期:  2024-02-04
  • 录用日期:  2024-03-07
  • 出版日期:  2024-10-25
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