基于动物实验和网络药理学的板参散防治蛋鸡脂肪肝综合征作用机制分析

Analysis of the mechanism of Banshensan in the prevention and treatment of fatty liver syndrome in laying hens based on animal experiments and network pharmacology

为探究板参散防治蛋鸡脂肪肝综合征的作用机制,试验使用高能低蛋白日粮饲喂蛋鸡造模,检测板参散对模型蛋鸡肝脏组织形态和肝脏抗氧化性的影响,然后采用网络药理学方法获得板参散防治脂肪肝综合征的靶点蛋白信息,并对其进行蛋白互作关系分析、GO功能注释、KEGG信号通路富集分析和分子对接分析。结果表明:与模型组相比,饲喂板参散的药物组蛋鸡肝细胞内脂肪小滴蓄积减少,肝细胞形态结构正常,肝细胞间隙清晰,肝脏丙二醛含量显著降低(P<0.05),总抗氧化能力显著提高(P<0.05);筛选得到板参散中β-谷甾醇、木犀草素、丹参酮ⅡA及隐丹参酮等89个活性成分,其主要作用于丝氨酸/苏氨酸蛋白激酶Akt(AKT1)、肿瘤抗原P53(TP53)和信号转导子与转录激活子3(STAT3)等94个潜在蛋白靶点,β-谷甾醇、木犀草素、丹参酮ⅡA与关键靶点AKT1、TP53均具有良好的分子对接活性;核心靶点主要富集于细胞凋亡过程的负调控、细胞凋亡过程的正调控和RNA聚合酶Ⅱ启动子转录的正调控等生物学过程,大分子复合物、核质、受体复合物等细胞组分,酶结合和相同蛋白结合等分子功能,以及脂质和动脉粥样硬化、PI3K/Akt信号通路、缺氧诱导因子信号通路等信号通路。说明板参散可通过多成分、多靶点、多通路机制防治脂肪肝。

In order to explore the mechanism of Banshensan compound in the prevention and treatment of fatty liver syndrome in laying hens,in the experiment,high-energy and low-protein diets were used to feed laying hens for the modeling,and the effects of Banshensan on the liver tissue morphology and liver antioxidant activity of the model laying hens were detected.Then,the network pharmacology method was used to obtain the target information of Banshensan for the prevention and treatment of fatty liver syndrome,and the protein-protein interaction analysis,GO function annotations,KEGG signaling pathway enrichment analysis and molecular docking analysis were carried out.The results showed that,compared with the model group,the accumulation of fat droplets in the hepatocytes of the drug group was reduced;the morphological structure of hepatocytes was normal;the hepatocyte space was clear;the content of malondialdehyde in the liver was significantly reduced(P<0.05),and the total antioxidant capacity was significantly increased(P<0.05).A total of 89 active ingredients,including β-sitosterol,luteolin,tanshinone Ⅱ A and cryptotanshione were screened and obtained,which played a role in 94 potential targets such as serine/threonine protein kinase Akt(AKT1),tumor antigen P53(TP53) and signal transducer and activator of transcription 3(STAT3).β-sitosterol,luteolin and tanshinone Ⅱ A had good molecular docking activities with key targets AKT1 and TP53.The core targets were mainly enriched in biological processes such as the negative regulation of apoptosis,the positive regulation of apoptosis and the positive regulation of RNA polymerase Ⅱ promoter transcription,cellular components such as macromolecular complexes,nucleoplasm and receptor complexes,molecular functions such as enzyme binding and same protein binding,as well as lipid and atherosclerosis,PI3K/Akt signaling pathway,HIF-1 signaling pathway and others.The results suggested that Banshensan could prevent and treat fatty liver through multi-component,multi-target and multi-pathway mechanisms.