中国药理学与毒理学杂志2025，Vol.39Issue(11)：829-838,10.DOI:10.3867/j.issn.1000-3002.2025.08459
脉络宁通过改善肿瘤微环境对Hepa1-6异位移植小鼠肝细胞癌生长的抑制作用及可能机制
Mailuoning inhibits hepatocellular carcinoma growth by improving tumor microenvironments in Hepa1-6 tumor-bearing mice
摘要
Abstract
OBJECTIVE To explore the inhibitory effect of Mailuoning(MLN)on ectopic hepatocellular carcinoma(HCC)growth and the potential mechanism.METHODS ①C57BL/6Jmale mice were inoc-ulated with Hepa1-6 cells(1×106)in the axilla to establish ectopic HCC models and randomly divided into 4 groups:model group,model+MLN 5 g·kg-1 group,model+MLN 15 g·kg-1 group and model+sorafenib(SRF)30 mg·kg-1 group.The drug administration groups were intragastrically given corresponding doses of drugs for 14 consecutive days,and body mass and tumor volume were monitored every 2 days during this period.At the end of the animal experiment,mice were sacrificed and tumors were harvested.The anti-tumor effect of MLN was evaluated via tumor illustration,tumor mass and tumor size.Bioinformatic analyses were conducted to identify key biological processes in hepatocellular carci-noma.Sirius red staining was used to assess collage fiber content in tumors of mice.Quantitative real-time polymerase chain reaction(RT-qPCR)test was performed to detect mRNA expressions of extra-cellular matrix components including collagen typeⅠalpha 1 chain(Col1a1),Col3a1 and fibronectin,epithe-lial-mesenchymal transition markers like E-cadherin,N-cadherin,α-smooth muscle actin(α-SMA)and vimentin,and immune checkpoint-related molecules such as programmed death-1(PD-1),programmed death-ligand 1(PD-L1)and B7 homologue 3(B7-H3).Protein levels of E-cadherin,α-SMA,and PD-L1 were subsequently detected by Western blotting.② HepG2 cells were incubated with the vehicle,β-ecdy-sone,harpagide,harpagoside,chlorogenic acid,isochlorogenic acid A,isochlorogenic acid C,luteoloside,macranthoidin B and dipsacoside B(10 μmol·L-1)for 24 h,respectively.After incubation,the expression of Col1a1 mRNA was measured to assess the impact of MLN active ingredients on COL1A1 produc-tion.Further more,HepG2 cells were incubated with the vehicle,β-ecdysone,chlorogenic acid,luteolo-side or macranthoidin B(10 μmol·L-1)for 24 h before the COL1A1 protein level was assessed.RESULTS① Bioinformatic analysis results showed that extracellular matrix and fibroblast proliferation related genes were notably changed in HCC patients.Compared with the model group,the mass and volume of tumors were significantly reduced in the model+MLN 15 g·kg-1 group,so was the positive area of sirius red staining in tumor tissues.In the model+MLN 15 g·kg-1 group,the mRNA expression of E-cadherin was upregulated while those of Col1a1,Col3a1,fibronectin,N-cadherin,α-SMA,vimentin,PD-L1 and B7-H3 were downregulated compared to the model group.Western blotting results revealed that the E-cadherin protein level in HCC tissues in the model+MLN 15 g·kg-1 group increased while the protein levels of α-SMA and PD-L1 in HCC tissues in both the model+MLN 5 g·kg-1 and model+MLN 15 g·kg-1 group decreased compared to the model group.②MLN active ingredients-β-ecdysone,chlorogenic acid,luteoloside,macranthoidin B-were effective at decreasing the mRNA and protein expressions of Col1a1 in HepG2 cells compared with the control group.CONCLUSIN MLN can ameliorate ectopic HCC growth by inhibiting epithelial-mesenchymal transition,decreasing extracellular matrix deposition,reducing immune checkpoint-related molecule PD-L1 expressions and improving immune microenvironments.关键词
脉络宁/肝细胞癌/细胞外基质/上皮-间充质转化/免疫检查点/肿瘤微环境Key words
Mailuoning/hepatocellular carcinoma/extracellular matrix/epithelial-mesenchymal transition/immune checkpoint/tumor microenvironment分类
医药卫生引用本文复制引用
周思妍,吴泽奇,柯创,陆宾,黄镇林,季莉莉..脉络宁通过改善肿瘤微环境对Hepa1-6异位移植小鼠肝细胞癌生长的抑制作用及可能机制[J].中国药理学与毒理学杂志,2025,39(11):829-838,10.基金项目
国家重点研发计划(2024YFC3506600) National Key R&D Program of China(2024YFC3506600) （2024YFC3506600）
