玉屏风散对肝癌微环境中血管生成的调节机制

Regulation Mechanism of Yupingfeng San on Angiogenesis in Liver Cancer Microenvironment

目的阐明玉屏风散通过抑制胸腺基质淋巴细胞生成素(TSLP)和信号转导因子(STAT3)的活化,调控肝癌微环境的免疫状态抑制微血管的形成从而发挥抗肝癌效应的机制。方法采用Hepa1-6原位肝癌小鼠模型,灌胃给予玉屏风散药液(20、30、40 g/kg),给药2周后处死小鼠。剥离瘤体称质量并计算抑瘤率;ELISA法检测肿瘤组织和癌旁组织上清液中VEGF、TSLP、TSLPR的表达水平;免疫组化法检测微血管密度MVD的表达情况;Western blot法检测STAT3和p-STAT3的蛋白表达情况。结果与模型组比较,不同浓度的玉屏风散(20、30、40 g/kg)可明显抑制肿瘤的生长(P<0.05~0.01),且呈剂量依赖,抑瘤率分别为26.37%、35.89%和56.01%;玉屏风散组肿瘤组织和癌旁组织的MVD和VEGF水平明显降低(P<0.05~0.01);不同浓度的玉屏风散可降低TSLP/TSLPR和p-STAT3/STAT3的表达,并且呈剂量依赖性(P<0.05~0.01)。结论玉屏风散可通过抑制TSLP表达,进而减弱TSLP-STAT3信号通路的激活,抑制肝癌微环境中血管的形成,发挥抗肝癌作用,该研究为延长肝癌患者生存和开发理想治疗药物靶点的新策略提供了实验依据。

OBJECTIVE To clarify that Yupingfeng San(YPFS)can inhibit the formation of microvessels by inhibiting the activation of thymic stromal lymphopoietin(TSLP)and signal transduction factor(STAT3),and exert its anti-liver effect.METHODS Hepa1-6 orthotopic liver cancer mouse model was used,and YPFS solution(20,30,40 g/kg)was intragastrically administered.The mice were sacrificed two weeks after administration.The tumors were weighed and the tumor inhibitory rate was calculated.The expression levels of VEGF,TSLP and TSLPR in tumor tissues and adjacent tissues were detected by ELISA.The expression of MVD in microvessel density was detected by immunohistochemistry.The protein expression of STAT3 and p-STAT3 was detected by Western blot.RESULTS Compared with the model group,different concentrations of YPFS(20,30 and 40 g/kg)groups significantly inhibited tumor growth in a dose-dependent manner(P<0.05,P<0.01),the tumor inhibitory rate was 26.37%,35.89%and 56.01%,respectively;the MVD and VEGF levels of tumor tissues and adjacent tissues in YPFS groups were significantly reduced(P<0.05,P<0.01);further studies showed that the expression of TSLP/TSLPR and p-STAT3/STAT3 in YPFS-treated groups could be reduced in dose-dependent manner(P<0.05,P<0.01).CONCLUSION YPFS attenuates the activation of TSLP-STAT3 signaling pathway by inhibiting the expression of TSLP,thereby inhibits the formation of blood vessels in the hepatic microenvironment and exerts anti-HCC effect.The study provideds experimental basis for new strategies for prolonging the survival of liver cancer patients and developing ideal therapeutic drugs targets.