荔枝核总黄酮通过抑制TLR4-TAK1抗大鼠肝纤维化的机制分析  

Mechanism of total flavones from litchi seed against liver fibrosisin rats by inhibiting TLR4-TAK1

作　　者：覃田田 黄盼玲 胡兴兰 罗伟生[1] 李宇清[1,3] QIN Tiantian;HUANG Panling;HU Xinglan;LUO Weisheng;LI Yuqing(Key Laboratory of Chinese Medicine Pharmacology of Education Department of Guangxi Zhuang Autonomous Region,Guangxi University of Chinese Medicine,Nanning 530200,China;Hechi Traditional Chinese Medicine Hospital,Hechi 547099,China;School of Innovation and Entrepreneurship,Guangxi University of Chinese Medicine,Nanning 530200,China)

机构地区：[1]广西中医药大学广西高校中药药理重点实验室,广西南宁530200 [2]河池市中医医院,广西河池547099 [3]广西中医药大学创新创业学院,广西南宁530200

出　　处：《药学研究》2024年第5期417-421,共5页Journal of Pharmaceutical Research

基　　金：国家自然科学基金项目(No.82160834);广西壮瑶药重点实验室开放课题(No.GXZYZZ2022-15);广西中医药大学桂派杏林拔尖人才项目(No.2022C007);广西中医药大学岐黄工程高层次人才团队培育项目(No.202405):中药学广西一流学科(No.桂教科研〔2022〕1号)。

摘　　要：目的探讨荔枝核总黄酮对四氯化碳(CCl 4)诱导的肝纤维化模型的抗肝纤维化作用机制。方法利用40%CCl 4-花生油溶液背部皮下注射构建肝纤维化大鼠模型,将造模成功的大鼠分为模型组、扶正化瘀组、荔枝核总黄酮低、中、高剂量组,另设空白组。连续灌胃给药8周后,取肝左大叶进行HE染色,观察肝脏组织病理变化;RT-qPCR法检测各组大鼠肝脏ColⅠ、ColⅢ、α-SMA mRNA的表达水平;并通过Western blotting检测大鼠肝脏Toll样受体-4(TLR4)、转化生长因子β激活激酶1(TAK1)、磷酸化转化生长因子β激活激酶1(P-TAK1)、氨基末端蛋白激酶(JNK)、磷酸化氨基末端蛋白激酶(P-JNK)的表达情况。结果肝组织HE染色显示,与肝纤维化模型大鼠比较,荔枝核总黄酮各剂量均能不同程度的改善大鼠肝脏炎症浸润情况。肝纤维化相关因子检测显示,荔枝核总黄酮各剂量肝脏ColⅠ、ColⅢ与α-SMA mRNA表达下降(P<0.05或P<0.01)。Western blotting检测结果显示,与模型组比较,荔枝核总黄酮低、中剂量组肝脏TLR4、P-TAK1蛋白表达下降(P<0.05),低剂量组的肝脏P-JNK蛋白也降低(P<0.05)。结论荔枝核总黄酮能减轻CCl 4诱发的大鼠肝纤维化,此作用可能与抑制肝脏TLR4-TAK1通路有关。Objective To explore the mechanism of anti-hepatic fibrosis effect of total flavones from litchi seed on carbon tetrachloride-induced hepatic fibrosis model.Methods Rat model of hepatic fibrosis was constructed by subcutaneous dorsal injection of 40%carbon tetrachloride-peanut oil solution,and the rats successfully modeled were divided into model group,Fuzheng Huayu group,and total flavones from litchi seed low-,medium-,and high-dose groups,and normal group was also set up.After 8 weeks of continuous gavage administration,the left lobe of the liver was taken for HE staining to observe the histopathological changes in the liver;The expression levels of ColⅠ,ColⅢ,andα-SMA mRNA in the liver of the rats in each group were detected by RT-qPCR;And the expression of TLR4,TAK1,P-TAK1,JNK,and P-JNK in the liver of the rats was detected by Western blotting.Results HE staining of liver tissues showed that all doses of total flavones from litchi seed improved the inflammatory infiltration of rat liver to different degrees when compared with that of rats with liver fibrosis model.The detection of liver fibrosis-related factors showed that the expression of ColⅠ,ColⅢandα-SMA mRNA in the livers of rats at all doses of total flavones from litchi seed was significantly decreased(P<0.05 or P<0.01).The results of Western blotting showed that the expression of liver TLR4 and P-TAK1 proteins in the low and medium dose groups of total flavones from litchi seed was significantly decreased(P<0.05)compared with that in the model group,and the hepatic P-JNK protein in the low dose group was also significantly decreased(P<0.05).Conclusion Total flavones from litchi seed attenuated carbon tetrachloride-induced hepatic fibrosis in rats,and this effect may be related to the inhibition of hepatic TLR4-TAK1 pathway.

关 键 词：荔枝核总黄酮 肝纤维化 TOLL样受体-4 转化生长因子β激活激酶1 

分 类 号：R285.5[医药卫生—中药学]