藤茶总黄酮通过调控TGF-β1/p38MAPK通路对肝纤维化大鼠的作用及机制研究  

Study on the eff ect and mechanism of the total fl avonoids of Ampelopsis grossedentata on the hepatic fi brosis in rats by regulating the TGF-β1/p38MAPK pathway

作　　者：李雯 李木松 王利兵[3] LI Wen;LI Musong;WANG Libing(Department of Pharmacy,Handan Third Hospital,Handan 056000,China;Department of Hepatology,Baoding People’s Hospital,Baoding 071000,China;Department of Hepatobiliary Surgery,Tangshan Workers’Hospital,Tangshan 063000,China)

机构地区：[1]邯郸市第三医院药剂科,河北邯郸056000 [2]保定市人民医院肝五科,河北保定071000 [3]唐山市工人医院肝胆外科,河北唐山063000

出　　处：《吉林中医药》2021年第8期1074-1079,共6页Jilin Journal of Chinese Medicine

基　　金：2020年度河北省医学科学研究课题计划(20201490)。

摘　　要：目的探讨藤茶总黄酮(TF)通过调控转化生长因子β1/p38-促分裂原活化蛋白激酶(TGF-β1/p38MAPK)通路对肝纤维化大鼠的作用及机制。方法65只Wistar大鼠,随机分为5组,对照组(灌胃生理盐水)、模型组(灌胃生理盐水)、藤茶总黄酮低剂量组(灌胃75 mg/kg)、藤茶总黄酮高剂量组(灌胃300 mg/kg)、激动剂组(灌胃300 mg/kg藤茶总黄酮加腹腔注射2 mg/kg anisomycin),每组13只,除对照组外,余组采用皮下注射25%CCl4建立肺纤维化模型,给予相应药物干预5周。检测5组血清ALT、AST、肝组织ColⅠ、ColⅢ水平,HE染色观察肝组织病理改变,实时荧光定量PCR(RT-qPCR)检测肝组织中TGF-β1、p38MAPK、α平滑肌肌动蛋白(α-SMA)、基质金属蛋白酶-9(MMP-9)mRNA相对表达量,Western blot法检测肝组织中TGF-β1、p38MAPK、p-p38MAPK、α-SMA、MMP-9蛋白相对表达量。结果与模型组比较,藤茶总黄酮低剂量组、藤茶总黄酮高剂量组及激动剂组血清ALT、AST及肝组织ColⅠ、ColⅢ水平均降低,其中藤茶总黄酮高剂量组ALT、AST、ColⅠ低于藤茶总黄酮低剂量组和激动剂组(P<0.05)。HE染色结果显示模型组肝组织结缔严重,肝脏明显纤维化,藤茶总黄酮低剂量组、藤茶总黄酮高剂量组、激动剂组肝纤维化程度明显减轻,其中藤茶总黄酮高剂量组肝脏恢复程度最佳。与模型组比较,藤茶总黄酮低剂量组、藤茶总黄酮高剂量组及激动机组TGF-β1、α-SMA mRNA和蛋白相对表达量均降低,p-p38MAPK蛋白相对表达量降低,MMP-9 mRNA和蛋白相对表达量均升高(P<0.05),其中藤茶总黄酮高剂量组TGF-β1、α-SMA mRNA和蛋白相对表达量及p-p38MAPK蛋白相对表达量均低于藤茶总黄酮低剂量组及激动剂组,MMP-9 mRNA和蛋白相对表达量均高于藤茶总黄酮低剂量组及激动剂组(P<0.05)。结论藤茶总黄酮可显著抑制大鼠肝纤维化,其调控机制可能与TGF-β1/p38MAPK信号通路有关。Objective To investigate the effect and mechanism of the total flavonoids(TF)of Ampelopsis grossedentata on the hepatic fi brosis in rats by regulating the transforming growth factorβ1/p38-mitogen-activated protein kinase(TGF-β1/p38MAPK)pathway.Methods A total of 65 Wistar rats were randomly divided into 5 groups:the control group(normal saline by gavage),the model group(normal saline by gavage),the TF low-dose group(75 mg/kg TF by gavage),the TF high-dose group(300 mg/kg TF by gavage),the agonist group(300 mg/kg TF by gavage+intraperitoneal injection of 2 mg/kg anisomycin),with 13 rats in each group.Except the control group,the pulmonary fibrosis model was established by the subcutaneous injection of 25%CCl4 in other groups,and a corresponding drug intervention was given for 5 weeks.The levels of ALT,AST in serum,of Col I and Col III in liver tissues were detected in the 5 groups.The pathological changes of liver tissues were observed by HE staining.The relative expression levels of TGF-β1,p38MAPK,αsmooth muscle actin(α-SMA)and matrix metalloproteinase-9(MMP-9)mRNA were detected by the real-time fl uorescent quantitative PCR(RT-qPCR).The relative expression levels of TGF-β1,p38MAPK,α-SMA and MMP-9 in liver tissues were detected by Western blot.Results Compared with the model group,the levels of serum ALT,AST,and liver tissue Col I,Col III in the TF low-dose group,TF high-dose group,and agonist group were decreased,and ALT,AST,Col I in the TF high-dose group were lower than those in the TF low-dose group and agonist group(P<0.05).The results of HE staining showed that the liver tissues of the model group were severely connected,and the livers were obviously fi brotic.While the degree of liver fi brosis was signifi cantly lowered in the low-dose TF group,high-dose TF group and agonist group,and the high-dose TF group had the best degree of liver recovery.Compared with the model group,the relative expression levels of the TGF-β1,α-SMA mRNA and protein in the TF low-dose group,TF high-dose group and agonist

关 键 词：肝纤维化 藤茶总黄酮 转化生长因子Β1 p38-促分裂原活化蛋白激酶 

分 类 号：R575.2[医药卫生—消化系统]