基于生物信息分析及实验验证探讨赤芍-附子治疗慢加急性肝衰竭的作用机制
Mechanism of Paeoniae Radix Rubra and Aconiti Lateralis Radix Praeparata in Treatment of Acute-on-chronic Liver Failure Based on Bioinformation Analysis and Experimental Validation
田晓玲 1张彧 1杜珊 1伍梦思 1谭年花 1陈斌1
作者信息
- 1. 湖南中医药大学第一附属医院,长沙 410007
- 折叠
摘要
目的:通过网络药理学、分子对接及动物实验探究赤芍-附子治疗慢加急性肝衰竭(ACLF)的作用机制.方法:通过网络药理学获得赤芍-附子治疗ACLF的可能靶点和相关信号通路,并通过分子对接技术检验核心成分与对应关键靶点的结合活性.采用牛血清白蛋白皮下及尾静脉注射联合脂多糖(LPS)+D-氨基半乳糖(D-GalN)腹腔注射构建ACLF大鼠模型,实验设置正常组(NC)、模型组、赤芍-附子组(5.85 g·kg-1)、促肝细胞生长颗粒组(HGFG,4.05g·kg-1).苏木素-伊红(HE)及马松(Masson)染色观察大鼠肝组织病理变化.酶联免疫吸附测定法(ELISA)检测白细胞介素-6(IL-6)、B细胞淋巴瘤-2(Bcl-2)、胱天蛋白酶-3(Caspase-3)、白蛋白(ALB)表达水平.实时荧光定量聚合酶链式反应(Real-time PCR)、蛋白免疫印迹法(Western blot)检测磷脂酰基醇3-激酶(PI3K)、蛋白激酶B(Akt)、磷酸化(p)-PI3K、p-Akt mRNA及蛋白表达水平.结果:网络药理学筛选得到赤芍-附子活性成分49种,作用靶点103个,ACLF疾病靶点3 317个,其中74个与赤芍-附子药物靶点重合.蛋白质-蛋白质相互作用(PPI)网络关键节点为Aktl、肿瘤坏死因子(TNF)、IL-6、Bcl-2、Caspase-3等;基因本体(GO)功能分析与京都基因与基因组百科全书(KEGG)富集分析筛选出多种信号通路,其中PI3K/Akt信号通路出现频率最高.分子对接显示,药物核心成分与对应关键靶点均具有较好的结合活性.动物实验证实,赤芍-附子可显著改善ACLF大鼠肝组织病理损伤,减少炎症因子的释放、肝细胞凋亡,并上调PI3K/Akt信号通路表达水平.结论:通过网络药理学-分子对接和体内实验,该研究证实了赤芍-附子减轻肝细胞炎性损伤、抑制肝细胞凋亡的作用效应,并初步探索了其具体机制,可能与参与调节PI3k/Ak信号通路有关.
Abstract
Objective:To explore the mechanism of action of Paeoniae Radix Rubra and Aconiti Lateralis Radix Praeparata(CSFZ)in the treatment of acute-on-chronic liver failure(ACLF)through network pharmacology,molecular docking,and animal experiments.Methods:Network pharmacology was used to identify potential targets and related signaling pathways for the treatment of ACLF with CSFZ.Molecular docking was used to examine the binding activity of the core components with corresponding key targets.An ACLF rat model was established by subcutaneous and tail vein injections of bovine serum albumin combined with lipopolysaccharide(LPS)+D-galactosamine(D-GalN)intraperitoneal injection.A normal control group(NC),a model group,a CSFZ group(CSFZ,5.85 g·kg-1),and a hepatocyte growth-promoting granule group(HGFG,4.05 g·kg-1)were set up in this study.Pathological changes in rat liver tissue were observed using hematoxylin and eosin(HE)and Masson staining.Enzyme-linked immunosorbent assay(ELISA)was used to detect the expression levels of interleukin-6(IL-6),B-cell lymphoma-2(Bcl-2),Caspase-3,and albumin(ALB).Real-time quantitative polymerase chain reaction(Real-time PCR)and Western blot were used to measure the mRNA and protein expression levels of phosphoinositide 3-kinase(PI3K),protein kinase B(Akt),phosphorylated PI3K(p-PI3K),and phosphorylated Akt(p-Akt).Results:Network pharmacology screening identified 49 active ingredients of CSFZ,103 action targets,and 3 317 targets related to ACLF.Among these,74 targets overlapped with CSFZ drug targets.Key nodes in the protein-protein interaction(PPI)network included Aktl,tumor necrosis factor(TNF),IL-6,Bcl-2,and Caspase-3.Gene Ontology(GO)functional analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis identified multiple signaling pathways,with the PI3K/Akt signaling pathway being the most frequent.Molecular docking showed that the core components of the drug exhibited good binding activity with the corresponding key targets.Animal experiments confirmed that CSFZ significantly improved liver tissue pathological damage in ACLF rats,reduced the release of inflammatory factors and liver cell apoptosis,and upregulated the expression levels of the PI3K/Akt signaling pathway.Conclusion:Through network pharmacology,molecular docking,and in vivo experiments,this study confirms the effect of CSFZ in reducing liver cell inflammatory damage and inhibiting liver cell apoptosis.The specific mechanism may be related to its involvement in regulating the PI3K/Akt signaling pathway.
关键词
慢加急性肝衰竭/赤芍/附子/磷脂酰基醇3-激酶(PI3K)/蛋白激酶B(Akt)/网络药理学Key words
acute-on-chronic liver failure/Paeoniae Radix Rubra/Aconiti Lateralis Radix Praeparata/phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt)/network pharmacology引用本文复制引用
出版年
2025
国家科技期刊平台
NSTL
维普
万方数据