膈下逐瘀汤对四氯化碳致大鼠肝纤维化的改善作用及机制初探  

Effect of Gexia Zhuyu Decoction on CCl_(4)-induced Liver Fibrosis in Rats and the Mechanism

作　　者：孙旭[1] 熊芬[1] 黄育生[2] 闫雪[1] 蔡庆群[1] 丘振文[1] SUN Xu;XIONG Fen;HUANG Yu-sheng;YAN Xue;CAI Qing-qun;QIU Zhen-wen(The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou Guangdong 510405;Lingnan Medical Research Center of Guangzhou University of Chinese Medicine,Guangzhou Guangdong 510405)

机构地区：[1]广州中医药大学第一附属医院,广东广州510405 [2]广州中医药大学岭南医学研究中心,广东广州510405

出　　处：《世界中西医结合杂志》2022年第10期1970-1973,1979,共5页World Journal of Integrated Traditional and Western Medicine

基　　金：广东省中医药管理局项目(20201094)。

摘　　要：目的探讨膈下逐瘀汤治疗四氯化碳(Carbon tetrachloride,CCl_(4))诱导大鼠肝纤维化(Hepatic fibrosis,HF)的可能作用机制。方法将48只成年SD雄性大鼠随机分为正常组、模型组、秋水仙碱对照组和膈下逐瘀汤组,每组各12只。除正常组外,其余3组采用四氯化碳改良法制备肝纤维化模型。大鼠首次腹腔注射40%CCl_(4)花生油溶液的第2天起,正常组和模型组灌胃生理盐水10 ml/kg,阳性对照组予秋水仙碱0.11 mg/kg,膈下逐瘀汤组予生药饮片浓煎液7.29 g/kg,1次/d,连续8周。Masson染色检测肝组织损伤及纤维化程度;免疫组化法检测大鼠肝组织中转化生长因子-β1(TGF-β1)、Wnt 1表达水平,实时聚合酶链反应(Real-time PCR)检测Wnt 4的相对表达量。结果正常组大鼠有少量胶原纤维出现在肝脏汇管区周围,未见纤维增生;模型组大鼠肝细胞排列紊乱,可见广泛纤维组织增生及假小叶形成,经秋水仙碱和膈下逐瘀汤干预后,各给药组胶原纤维增生情况明显减少。正常组仅有少量阳性标记,细胞质内见有少量的棕色颗粒。模型组阳性表达增强,可见大量棕黄色颗粒,秋水仙碱组和膈下逐瘀汤组阳性标记较模型组明显减少。与正常组比较,模型组TGF-β1、Wnt1蛋白表达及Wnt4 mRNA表达明显升高,差异有统计学意义(P<0.01);与模型组比较,膈下逐瘀汤组和秋水仙碱组TGF-β1、Wnt1蛋白表达及Wnt4 mRNA表达均降低,差异有统计学意义(P<0.05,P<0.01)。结论膈下逐瘀汤有效改善CCl_(4)致大鼠肝损伤,其作用机制可能与抗氧化应激、抑制炎症反应、阻断肝纤维化有关。Objective To investigate the underlying mechanism of Gexia Zhuyu Decoction(GZD)against the carbon tetrachloride(CCl_(4))-induced liver fibrosis in rats.Methods A total of 48 male adult SD rats were randomized into the normal group,model group,colchicine group,and GZD group,with 12 rats in each group.The modified CCl_(4)method was used to induce liver fibrosis in rats except the normal group.On the 2 nd day after the first intraperitoneal injection of 40%CCl_(4)in peanut oil,normal group and model group were given(i.g.)normal saline(10 mL/kg).Colchicine group received colchicine(0.11 mg/kg),and GZD group was treated with concentrated decoction of the decoction pieces for GZD at 7.29 g/kg,once a day for 8 consecutive weeks.Liver injury and fibrosis were observed based on Masson staining.Immunohistochemical method was used to detect the expression of TGF-β1 and Wnt1 in liver tissue,and real-time PCRto determine the expression of Wnt 4.Results Rats in the normal group showed a few collagenous fibers in the portal area of liver without fibrogenesis.Rats in the model group demonstrated disordered arrangement of hepatocytes,extensive fibrous hyperplasia,and pseudolobule formation.Colchicine and GZD significantly alleviated the proliferation of collagenous fibers.The normal group showed a few positive markers with a few brown granules in the cytoplasm.The model group displayed increase in positive expression and a large number of brown granules.The colchicine group and GZD group witnessed the significant reduction in positive markers as compared with the model group.The protein expressionof TGF-β1 and Wnt1 and expression of Wnt 4 mRNA in liver tissue were higher in the model group than in the normal group(P<0.01),and the protein expressionof TGF-β1 and Wnt1 and expression of Wnt4 mRNA were lower in thecolchicine group and GZD group than in the model group(P<0.05,P<0.01).Conclusion GZD can effectively improve the liver injury caused by CCl_(4)in ratsthrough anti-oxidative stress and inhibition of inflammation and liver

关 键 词：膈下逐瘀汤 肝纤维化 转化生长因子-Β Wnt蛋白 

分 类 号：R285.5[医药卫生—中药学]