扶脾柔肝颗粒对肝纤维化模型大鼠的改善作用机制研究
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篇名: 扶脾柔肝颗粒对肝纤维化模型大鼠的改善作用机制研究
TITLE: Mechanism of Improvement Effects of Fupi Rougan Granules on Hepatic Fibrosis Model Rats
摘要: 目的:研究扶脾柔肝颗粒(FRG)对肝纤维化模型大鼠的改善作用机制。方法:将大鼠随机分为空白组、模型组、秋水仙碱片组(化学药阳性对照,0.2mg/kg)、扶正化瘀胶囊组(中药阳性对照,0.415g/kg)和FRG低、中、高剂量组(20、40、80g/kg),除空白组和模型组各11只大鼠外(各取1只用于判断是否造模成功),其余每组10只。除空白组外,其余各组大鼠腹腔注射50%CCl4橄榄油溶液并灌胃30%乙醇以复制肝纤维化模型。造模成功后,各给药组大鼠灌胃相应药物,空白组和模型组大鼠灌胃等体积生理盐水,每天1次,连续4周。末次灌胃后,观察大鼠肝组织病理形态学变化;检测大鼠血清中透明质酸(HA)、层粘连蛋白(LN)、Ⅲ型前胶原(PCⅢ)、Ⅳ型胶原蛋白(ColⅣ)水平;检测大鼠肝组织中Beclin-1、LC3-Ⅱ蛋白表达水平;检测大鼠肝组织中蛋白激酶B(Akt)、腺苷酸活化蛋白激酶(AMPK)、哺乳动物雷帕霉素靶蛋白(mTOR)、p70核糖体蛋白S6激酶(p70S6K)的mRNA和蛋白表达水平。结果:与空白组比较,模型组大鼠肝小叶结构紊乱,纤维组织增生明显,部分有假小叶形成;血清中HA、LN、PCⅢ、ColⅣ水平和肝组织中Beclin-1、LC3-Ⅱ蛋白表达水平以及Akt、AMPK、mTOR、p70S6K的mRNA和蛋白表达水平均显著升高(P<0.01)。与模型组比较,FRG各剂量组大鼠肝组织损伤明显减轻,血清和肝组织中上述指标(FRG低剂量组LN、PCⅢ除外)水平均显著降低(P<0.05或P<0.01)。结论:FRG可改善大鼠肝纤维化,其作用机制可能与下调肝组织中自噬相关蛋白和Akt/AMPK/mTOR/p70S6K信号通路相关蛋白的表达有关。
ABSTRACT: OBJECTIVE:To study the mechanism of improvement effects of Fupi rougan granule (FRG)on hepatic fibrosis model rats. METHODS :The rats were randomly divided into blank group ,model group ,Colchicine tablet group (chemical positive control ,0.2 mg/kg),Fuzheng huayu capsule group (TCM positive control ,0.415 g/kg),FRG low-dose ,medium-dose and high-dose groups (20,40,80 g/kg),with 10 rats in each group ,except for 11 rats in blank group and model group (one rat was used to judge whether the modeling was successful ). Except for blank group ,other groups were given intraperitoneal injection of 50% CCl4 olive oil solution and intragastric administration of 30% ethanol to induce hepatic fibrosis model. After modeling , administration groups were given relevant medicine intragastrically ;blank group and model group were given constant volume of normal saline intragastrically ,once a day ,for consecutive 4 weeks. After last administration ,morphology changes of liver tissue in rats were observed. The serum levels of HA ,LN,PCⅢ and Col Ⅳ in rats were detected ,and protein expression of Beclin- 1 and LC3-Ⅱin liver tissue were also determined. mRNA and protein expression of Akt ,AMPK,mTOR,p70S6K were detected in liver tissues of rats. RESULTS :Compared with blank group ,the structure of hepatic lobules in the model group was disordered ,the proliferation of fibrous tissue was obvious ,and some pseudolobules were formed ;the serum levels of HA ,LN,PCⅢ and Col Ⅳ, the protein expression of Beclin- 1 and LC 3-Ⅱ in liver tissue as well as mRNA and protein expression of Akt ,AMPK,mTOR and p70S6K were increased significantly (P<0.01). Compared with model group ,the liver injury of rats in FRG groups was significantly relieved ,and the levels of the above indexes in serum and liver tissue (except for LN and PC Ⅲ in FRG low-dose group) were significantly reduced (P<0.05 or P<0.01). CONCLUSIONS :FRG can improve hepatic fibrosis in rats ,the mechanism of which may be associated with down-regulating the expression of autophagy associated protein and Akt/AMPK/mTOR/ p70S6K signaling pathway related protein.
期刊: 2021年第32卷第21期
作者: 安祯祥,何远利,唐东昕,黄丹,王敏,王芳
AUTHORS: AN Zhenxiang,HE Yuanli,TANG Dongxin,HUANG Dan,WANG Min,WANG Fang
关键字: 扶脾柔肝颗粒;肝纤维化;自噬;Akt/AMPK/mTOR/p70S6K信号通路;大鼠
KEYWORDS: Fupi rougan granule ;Hepatic fibrosis ;Autophagy;Akt/AMPK/mTOR/p70S6K signaling pathway ;Rats
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