一贯煎对M1型骨髓巨噬细胞治疗肝硬化大鼠模型效果的影响及其机制分析  

Effect of Yiguan Decoction on the efficacy of M1 bone marrow-derived macrophages in treatment of liver cirrhosis rats and its mechanism

作　　者：宗梦瑶 简迅 王丹阳 许燕楠 郑欣瑞 邢飞飞 陈高峰[1] 陈佳美[1] 刘平[1] 慕永平[1] ZONG Mengyao;JIAN Xun;WANG Danyang;XU Yannan;ZHENG Xinrui;XING Feifei;CHEN Gaofeng;CHEN Jiamei;LIU Ping;MU Yongping(Key Laboratory of Liver and Kidney Diseases,Ministry of Education,Institute of Liver Diseases,Shuguang Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai Key Laboratory of Traditional Chinese Clinical Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属曙光医院,上海市中医药研究院肝病研究所,肝肾疾病病证教育部重点实验室,上海市中医临床重点实验室,上海201203

出　　处：《临床肝胆病杂志》2024年第8期1612-1619,共8页Journal of Clinical Hepatology

基　　金：国家自然科学基金面上项目(81874390);上海市科委自然科学基金面上项目(21ZR1464100);上海市科委2022年度“科技创新行动计划”生物医药科技支撑专项(22S11901700);上海市临床重点专科建设项目(shslczdzk01201)。

摘　　要：目的观察一贯煎对M1型骨髓巨噬细胞(M1-BMDM)治疗2-AAF/CCl4诱导的大鼠肝硬化作用的影响与机制。方法分离BMDM,脂多糖诱导分化为M1-BMDM。雄性Wistar大鼠随机分为正常组(n=5)和造模组(n=45)。造模组大鼠予以50%CCl4每周2次皮下注射。第7周开始,将造模大鼠随机分为模型组(M组)、一贯煎组(YGJD组)、M1-BMDM组、M1-BMDM+YGJD组、索拉非尼组(SORA组),改用30%CCl4皮下注射以维持肝硬化进展,同时以2-AAF灌胃,且饮水中加入CCR2抑制剂,分组干预,9周末取材。观察血清肝功能、肝组织病理、肝组织羟脯氨酸(Hyp)含量、肝星状细胞活化、肝纤维化及炎症相关因子、Wnt信号通路相关分子表达水平等。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果与M组比较,M1-BMDM+YGJD组大鼠血清ALT、AST、TBil水平均明显降低(P值均<0.05),Alb含量显著增加(P<0.05);与M1-BMDM组比较,M1-BMDM+YGJD组血清TBil含量显著降低(P<0.05)、Alb含量显著升高(P<0.05)。与M1-BMDM组比较,M1-BMDM+YGJD组CD68、TNF-α表达显著降低(P<0.05)。与M1-BMDM组比较,M1-BMDM+YGJD组Hyp含量及天狼星红胶原阳性染色面积均显著降低(P值均<0.05)。M1-BMDM+YGJD组非经典Wnt信号通路分子Wnt5a mRNA和蛋白表达均显著低于M1-BMDM组(P值均<0.05),Fzd2 mRNA表达水平显著低于M1-BMDM组(P<0.05),且Wnt4、Wnt5b和Fzd3 mRNA表达水平亦显著降低(P值均<0.05),而经典Wnt信号通路分子β-catenin、LRP5、LRP6、Fzd5和TCF mRNA表达水平均无明显改变。结论一贯煎可提高M1-BMDM对2-AAF/CCl4诱导的大鼠肝硬化的治疗效应,其机制可能与抑制非经典Wnt5a/Fzd2信号通路有关,为中医药协同M1-BMDM治疗肝硬化提供了新思路。Objective To investigate the effect and mechanism of Yiguan Decoction(YGJD)on the efficacy of M1 bone marrowderived macrophages(M1-BMDMs)in the treatment of rats with liver cirrhosis induced by 2-AAF/CCl4.Methods BMDMs were isolated and induced into M1-BMDMs by lipopolysaccharide.A total of 50 male Wistar rats were randomly divided into normal group with 5 rats and model group with 45 rats.The rats for modeling were given subcutaneous injection of 50%CCl4 twice a week.Since week 7,the rats for modeling were randomly divided into model group(M group),YGJD group,M1-BMDM group,M1-BMDM+YGJD group,and sorafenib(SORA)group,and they were given subcutaneous injection of 30%CCl4 to maintain the progression of liver cirrhosis and intragastric administration of 2-AAF.CCR2 inhibitors were added to the drinking water,and each group was given the corresponding intervention.Related samples were collected at week 9.The rats were observed in terms of serum liver function parameters,liver pathology,hydroxyproline(Hyp)content in liver tissue,hepatic stellate cell activation,hepatic fibrosis and inflammation factors,and the expression levels of molecules associated with the Wnt signaling pathway.A one-way analysis of variance 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 M group,the M1-BMDM+YGJD group had significant reductions in the serum levels of alanine aminotransferase,aspartate aminotransferase,and total bilirubin(TBil)(all P<0.05)and a significant increase in the content of albumin(Alb)(P<0.05),and compared with the M1-BMDM group,the M1-BMDM+YGJD group had a significant reduction in the serum level of TBil(P<0.05)and a significant increase in the serum level of Alb(P<0.05).Compared with the M1-BMDM group,the M1-BMDM+YGJD group had significant reductions in the expression levels of CD68 and TNF-α(P<0.05).Compared with the M1-BMDM group,the M1-BMDM+YGJD group had significant reductions

关 键 词：肝硬化 巨噬细胞 一贯煎 WNT信号通路 大鼠 Wistar 

分 类 号：R285.5[医药卫生—中药学]