基于网络药理学及分子对接方法探讨当归补血汤治疗慢性肾脏病的作用机制
The Mechanism of Angelicae Sinensis Decoction in the Treatment of Chronic Kidney Disease Based on Network Pharmacology and Molecular Docking
胡雅玲 1刘爽 2张紫媛 2刘文媛 2刘宇翔 3张晓东 2方敬爱2
作者信息
- 1. 山西医科大学第一医院 (太原 030001);山西医科大学 (太原 030001)
- 2. 山西医科大学第一医院 (太原 030001)
- 3. 山西医科大学 (太原 030001)
- 折叠
摘要
目的:基于网络药理学及分子对接方法分析当归补血汤治疗慢性肾脏病的作用机制,构建慢性肾衰竭大鼠模型进行实验验证.方法:利用数据库筛选当归补血汤活性成分及靶点、慢性肾脏病疾病靶点,取二者交集,构建蛋白互作(PPI)网络模型,对交集靶点进行GO与KEGG通路富集分析,构建药物-疾病靶点-通路网络图,得到当归补血汤治疗慢性肾脏病的关键靶点.对药物核心成分和疾病靶蛋白进行分子对接.构建慢性肾衰竭大鼠模型,运用免疫组化及 Western Blotting方法对核心靶点进行验证.结果:(1)当归补血汤治疗慢性肾脏病的潜在靶点涉及氧化应激、细胞粘附、凋亡等生物学过程,涉及蛋白激酶复合物、细胞外基质等细胞组成,涉及蛋白激酶活性、DNA结合转录因子结合、细胞因子受体结合等分子功能,涉及NF-κB信号通路、FoxO信号通路等多种信号途径.(2)筛选关键基因SIRT1、TLR4、MCP-1 与当归补血汤主要活性成分黄芪甲苷、毛蕊异黄酮、槲皮素进行分子对接,配体化合物均能良好的包埋在受体靶蛋白的活性口袋中.(3)构建慢性肾衰竭大鼠模型,给予当归补血汤结肠透析治疗,与正常组相比,CRF模型组肾组织SIRT1、Claudin-1、ZO-1 表达降低,TLR4、NF-κBp65、MCP-1 表达升高,给予当归补血汤结肠透析干预后,SIRT1、Claudin-1、ZO-1 表达升高,TLR4、NF-κBp65、MCP-1 表达降低,差异有统计学意义(P<0.05).结论:当归补血汤可通过多靶点、多途径发挥治疗慢性肾脏病的作用,当归补血汤经结肠透析治疗可能通过激活SIRT1 抑制TLR4 受体在肾脏细胞的表达,抑制NF-κBp65 乙酰化,从而抑制下游炎症反应,并通过修复肾脏细胞紧密连接结构改善慢性肾衰竭大鼠肾组织损伤.
Abstract
Objective:Analyzed the mechanism of Angelicae Sinensis Decoction in the treatment of chronic kidney disease based on the methods of network pharmacology and molecular docking.The rat model of chronic renal failure was constructed to verify the analysis results in vivo.Methods:Using databases to screen the active components and targets of Angelicae Sinensis Decoction,download the chronic kidney disease targets,take the intersection of those targets,construct the protein interaction(PPI)network model,use metascape platform to enrich and analyze the GO and KEGG pathways of the intersection targets,construct the drug-dis-ease target-pathway network diagram,and obtain the key targets of Angelicae Sinensis Decoction in the treatment of chronic kidney disease.Run autodock Vina for molecular docking of drug core components and disease target proteins.The rat model of chronic renal failure was established,and the core targets were verified by immunohistochemistry and Western blotting.Results:(1)The potential targets of Angelicae Sinensis Decoction in the treatment of chronic kidney disease involved biological processes such as oxidative stress,cell adhesion and apoptosis,cell composition such as protein kinase complex and extracellular matrix,molecular functions such as protein kinase activity,DNA binding,transcription factor binding and cytokine receptor binding.It also involved NF-κB sig-nal pathway,FOXO signal pathway and other signal pathways.(2)The key targets SIRT1,TLR4 and MCP-1 were screened for mo-lecular docking with astragaloside Ⅳ,calycosin and quercetin,the main active components of Angelicae Sinensis Decoction.The lig-and compounds can be well embedded in the active pocket of the receptor target protein.(3)The rat model of chronic renal failure was established and treated with Angelicae Sinensis Decoction colon dialysis.Compared with the normal group,the expression of SIRT1,Claudin-1 and ZO-1 in renal tissue of CRF model group were decreased,the expressions of TLR4,NF-κBp65 and MCP-1 were increased.After colon dialysis intervention with Angelicae Sinensis Decoction,the expressions of SIRT1,Caudin-1 and ZO-1 were increased,TLR4,NF-κBp65 and MCP-1 were decreased,and the difference was statistically significant(P<0.05).Conclu-sion:Angelicae Sinensis Decoctio can play an important role in the treatment of chronic kidney disease through multiple targets and pathways.Angelicae Sinensis Decoctio may activating SIRT1 and inhibit the expression of TLR4 receptor in renal cells,inhibition NF-κ Bp65 acetylation so as to control the downstream inflammatory response and improve the renal tissue injury of CRF rats by re-pairing the tight junction structure of renal cells.
关键词
当归补血汤/慢性肾脏病/网络药理学/结肠透析/炎症反应Key words
Angelicae sinensis decoction/Chronic kidney disease/Network pharmacology/Colon dialysis/Inflammation引用本文复制引用
基金项目
山西省科技厅应用基础研究计划(青年基金)(201901D211484)
山西省科技厅国际合作项目(201803D421063)
出版年
2024
NETL
NSTL
万方数据