祛痰活血方调控LPS介导TLR4/Myd88/NF-κB信号通路抗NASH大鼠肝损伤的机制研究

Mechanism of Qutan Huoxue Formula on LPS-mediated TLR4/Myd88/NF-κB Signaling Pathway against Liver Injury for NASH Rats

目的:观察祛痰活血方调控LPS抑制TLR4/Myd88/NF-κB信号通路抗NASH大鼠肝损伤的作用。方法:30只SPF级雄性SD大鼠随机分为正常组、模型对照组和祛痰活血方组,正常组给予普通饲料,模型对照组和祛痰活血方组给予高脂饮食8周建立NASH模型,造模成功后正常组和模型对照组灌胃生理盐水,祛痰活血方组给予中药灌胃,每日1次,连续4周。观察大鼠肝脏组织病理,血清ALT、AST、TC、TG、LPS,以及肝组织TLR4、Myd88、NF-κB表达的变化。结果:(1)与正常组比较,模型对照组大鼠肝小叶结构紊乱,可见大量气球样变细胞,局部可见少量炎症细胞浸润,并呈现大小不一的脂滴,祛痰活血方组大鼠肝脏脂肪变程度及肝细胞炎症均较模型对照组明显改善;(2)与正常组比较,模型对照组ALT、AST、TC、TG及LPS含量均明显升高,祛痰活血方组ALT、AST、TC、TG及LPS水平较模型对照组明显下降;(3)与正常组比较,模型对照组TLR4、MyD88及NF-κBmRNA水平和蛋白表达均显著上调(P<0.05),与模型对照组比较,祛痰活血方组TLR4、MyD88及NF-κBmRNA水平和蛋白表达均显著下调(P<0.05)。结论:祛痰活血方可以减少LPS产生,抑制TLR4/MyD88/NF-κB信号通路从而改善NASH大鼠肝损伤。

Objective:To observe the effect of Qutan Huoxue Formula on LPS inhibiting TLR4/Myd88/NF-κB signaling pathway against liver injury for NASH rats.Methods:30 SPF male SD rats were randomly divided into a normal group,a model controlled group and a Qutan Huoxue Formula group,the normal group was given normal feed,the model controlled group and the Qutan Huoxue Formula group were given high-fat diet for 8 weeks to establish NASH model.After successful modeling,the normal group and the model controlled group were gavaged with normal saline,and the Qutan Huoxue Formula group was given gavage of traditional Chinese medicine once a day for 4 weeks.Observed the pathology of rat liver tissue,serum ALT,AST,TC,TG,LPS,and the changes of liver tissue TLR4,Myd88,NF-κB expression.Results:(1)Compared with the normal group,the liver lobule structure of the model controlled group rats was disordered,a large number of balloon-like metamorphic cells were seen,a small amount of inflammatory cell infiltration was seen locally,and fat droplets of varying sizes were shown.The degree of liver fatty change and hepatocyte inflammation in the Qutan Huoxue Formula group were significantly improved compared with the model controlled group;(2)Compared with the normal group,the levels of ALT,AST,TC,TG and LPS in the model controlled group were significantly increased,and the levels of ALT,AST,TC,TG and LPS in the Qutan Huoxue Formula group were significantly decreased compared with the model controlled group;(3)Compared with the normal group,the mRNA and protein expressions of TLR4,MyD88,and NF-κB in the model controlled group were significantly up-regulated(P<0.05),compared with the model group,the mRNA and protein expressions of TLR4 MyD88 and NF-B in the model controlled group were significantly down-regulated(P<0.05).Conclusion:Qutan Huoxue Formula can reduce LPS production,inhibit TLR4/MyD88/NF-κB signaling pathway and improve liver injury in NASH rats.