文献类型：期刊文献

中文题名：半夏泻心汤抑制巨噬细胞M2型极化增敏PD-1单抗治疗肝细胞癌

英文题名：Banxia Xiexin Decoction(半夏泻心汤)Inhibits Macrophage Polarization to M2 Phenotype to Sensitize Hepatocellular Carcinoma to PD-1 Monoclonal Antibody

作者：何玲[1,2];刘喜平[1];陈光顺[1,2];谢知慧[1,2];朱中博[1];杨玉洁[1]

第一作者：何玲

机构：[1]甘肃中医药大学甘肃省中药新产品创制工程实验室、甘肃省中医方药挖掘与创新转化重点实验室,兰州730000;[2]甘肃中医药大学中医临床学院,兰州730000

第一机构：甘肃中医药大学科研实验中心（甘肃省中医药标准化技术委员会秘书处）

年份：2025

卷号：41

期号：4

起止页码：21

中文期刊名：中药药理与临床

外文期刊名：Pharmacology and Clinics of Chinese Materia Medica

收录：;北大核心:【北大核心2023】；

基金：国家自然科学基金资助项目(编号:82260895);甘肃省高校教师创新基金项目(编号:2024A-085);甘肃中医药大学博导专项基金项目(编号:ZYXKBD-20220)。

语种：中文

中文关键词：半夏泻心汤;肝细胞癌;巨噬细胞极化;内质网应激相关蛋白/信号转导和转录激活因子6/过氧化物酶体增殖物受体信号通路;程序性死亡受体1

外文关键词：Banxia Xiexin Decoction(半夏泻心汤);Hepatocellular carcinoma;Macrophage polarization;CHOP/STAT6/PPAR-γsignaling pathway;Programmed death 1

摘要：目的:探讨半夏泻心汤增敏PD-1单抗治疗肝细胞癌的机制。方法:BALB/c小鼠随机分为模型对照组、PD-1单抗组、半夏泻心汤3.9、7.8、15.6 g/kg组、联合(PD-1+半夏泻心汤7.8 g/kg)组,采用H22肝癌细胞异位移植瘤建立小鼠动物模型,药物干预后,观察肿瘤组织病理形态变化;免疫组织化学法及流式细胞术检测肿瘤癌组织中CD206、CD163表达水平。建立M2巨噬细胞炎症模型,CCK-8法筛选半夏泻心汤含药血清干预M2型巨噬细胞最佳浓度;酶联免疫吸附法检测血清中IFN-γ、TNF-α、IL12p70及TGF-β含量;流式细胞术检测干预M2型巨噬细胞表型变化及细胞凋亡情况;Western blot及RT-PCR法检测CHOP、STAT6及PPAR-γ蛋白及mRNA表达。结果:与模型对照组比较,PD-1单抗组、半夏泻心汤组瘤指数、脾脏指数、TGF-β含量、CD163、CD206水平明显降低(P<0.05或P<0.01),TNF-α、IFN-γ以及IL12p70含量显著升高(P<0.01);与PD-1单抗组比较,半夏泻心汤7.8、15.6 g/kg组及联合组瘤指数、脾脏指数、TGF-β含量、CD163、CD206水平升高(P<0.05或P<0.01),TNF-α、IFN-γ及IL12p70含量显著降低(P<0.01)。与模型对照组比较,二甲双胍组CD206+CD163+百分率明显降低,CHOP、STAT6、PPAR-γ蛋白及mRNA表达明显下调(P<0.05或P<0.01),凋亡率明显增加(P<0.05);与二甲双胍组比较,半夏泻心汤含药血清组CD206+CD163+百分率明显升高,CHOP、STAT6、PPAR-γ蛋白及mRNA表达上调(P<0.05或P<0.01),凋亡率明显降低(P<0.05)。结论:半夏泻心汤可抑制小鼠H22肝癌细胞株诱导的肝细胞癌肿瘤组织生长,且增敏PD-1单抗疗效,其作用机制可能与通过CHOP/STAT6/PPAR-γ通路抑制巨噬细胞M2型极化有关。
Objective:To investigate the mechanism of Banxia Xiexin Decoction(半夏泻心汤)in sensitizing hepatocellular carcinoma to the PD-1 monoclonal antibody.Methods:BALB/c mice were randomized into model control,PD-1 monoclonal antibody,Banxia Xiexin Decoction(15.6,7.8,and 3.9 g/kg),and combination(PD-1+Banxia Xiexin Decoction 7.8 g/kg)groups.The mouse model of hepatocellular carci-noma was established with the H22 cell-derived xenograft.After drug intervention,the pathological changes of the tumors were observed,and immunohistochemistry and flow cytometry were employed to detect the expression levels of CD206 and CD163 in the tumor tissue.In addition,the M2 macrophage model of inflammation was established,and the cell-counting kit-8(CCK-8)assay was employed to screen the optimal concentration of Banxia Xiexin Decoction-containing serum for treating M2 macrophages.Enzyme-linked immunosorbent assay was used to measure the levels of interferon(IFN)-γ,tumor necrosis factor(TNF)-α,interleukin-12p70(IL12p70),and TGF-βin the serum.Flow cy-tometry with Annexin V/propidium iodide was used to detect the changes in the phenotypes and apoptosis of M2 macrophages in each group.The protein and mRNA levels of C/EBP homologous protein(CHOP),signal transducer and activator of transcription 6(STAT6),and perox-isome proliferator-activated receptor(PPAR)-γwere determined by Western blotting and RT-PCR,respectively.Results:Compared with the model control group,the PD-1 monoclonal antibody group showed decreases in the tumor index,spleen index,and levels of TGF-β,CD163,and CD206(P<0.05 or P<0.01)and elevations in the levels of TNF-α,IFN-γ,and IL12p70(P<0.01).Compared with the PD-1 monoclonal antibody group,the Banxia Xiexin Decoction(15.6 and 7.8 g/kg)groups and the combination group showed increased tumor index,spleen index,and levels of TGF-β,CD163,and CD206(P<0.05 or P<0.01),and lowered levels of TNF-α,IFN-γ,and IL12p70(P<0.01).Com-pared with the model control group,the metformin group showed decreased CD206+CD163+percentage,down-regulated protein and mRNA lev-els of CHOP,STAT6,and PPAR-γ(P<0.05,P<0.01),and increased apoptosis rate(P<0.05).Compared with the metformin group,the Banxia Xiexin Decoction containing-serum group showed increased CD206+CD163+percentage,up-regulated protein and mRNA levels of CHOP,STAT6,and PPAR-γ(P<0.05,P<0.01),and reduced apoptosis rate(P<0.05).Conclusion:Banxia Xiexin Decoction can inhibit the growth of hepatocellular carcinoma induced by mouse H22 cell-derived xenograft and enhance the therapeutic efficacy of the PD-1 mono-clonal antibody by inhibiting the macrophage polarization to the M2 phenotype via the CHOP/STAT6/PPAR-γpathway.