β-谷甾醇对增生性瘢痕成纤维细胞作用机制的网络药理学分析  

Mechanism of beta-sitosterol on hypertrophic scar fibroblasts:an analysis based on network pharmacology

作　　者：左俊 马少林[1] Zuo Jun;Ma Shaolin(Department of Plastic Surgery,The First Affiliated Hospital of Xinjiang Medical University,Urumqi 830000,Xinjiang Uygur Autonomous Region,China)

机构地区：[1]新疆医科大学第一附属医院整形科,新疆维吾尔自治区乌鲁木齐市830000

出　　处：《中国组织工程研究》2024年第2期216-223,共8页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金地区基金项目(81760345),项目负责人:马少林:湖南省自然科学基金青年基金项目(2021JJ40487),项目负责人:左俊。

摘　　要：背景:目前针对增生性瘢痕有效的预防和治疗措施仍然有限。而大多数植物中草药不良反应小且来源丰富,为预防和治疗增生性瘢痕提供了新的思路和方法。目的:通过网络药理学和分子对接技术探讨植物来源β-谷甾醇作用于增生性瘢痕成纤维细胞的潜在分子机制,并通过细胞学实验进行初步验证。方法:通过网络药理学方法,利用相关数据库及软件筛选β-谷甾醇的药物靶点,获取增生性瘢痕相关疾病靶点,取交集得到β-谷甾醇作用于增生性瘢痕的潜在作用(交集)靶点,使用Cytoscape软件和STRING数据库构建“药物-靶点-疾病”网络和蛋白质互作网络(PPI),同时筛选出PPI中核心作用靶点。通过DAVID数据库对交集靶点进行GO生物学功能和KEGG通路富集分析,结合文献分析进一步确立与交集靶点关系密切的信号通路及核心靶点基因,应用AutoDock软件对β-谷甾醇和核心靶点蛋白进行分子对接。采用体外细胞实验验证β-谷甾醇对人增生性瘢痕成纤维细胞增殖、凋亡、细胞周期分布和核心靶点基因mRNA表达的影响。结果与结论:①β-谷甾醇和增生性瘢痕的交集靶点共56个,PPI网络中筛选出10个核心靶点:酪氨酸激酶(SRC)、丝裂原活化蛋白激酶3(MAPK3)、半胱氨酸蛋白酶3(CASP3)、载脂蛋白E(APOE)、雌激素受体1(ESR1)、固醇调节元件结合转录因子1(SREBF1)、过氧化物酶体增殖物激活受体α(PPARα)、C-反应蛋白(CRP)、细胞间黏附分子1(ICAM1)和过氧化氢酶(CAT)。②结合文献报道和KEGG通路、GO功能分析结果,进一步认定MAPK信号通路与交集靶点关系密切,确定MAPK3(即为ERK1-MAPK)、CASP3、P53和肿瘤坏死因子为其核心靶点。③分子对接结果提示β-谷甾醇与核心靶点蛋白结合良好。④细胞实验结果表明,100μmol/L的β-谷甾醇能抑制增生性瘢痕成纤维细胞增殖,降低线粒体膜电位、诱导细胞凋亡(P<0.01),使G1期细胞占�BACKGROUND:At present,effective preventive and therapeutic measures for hypertrophic scar are still limited.In contrast,most of botanical herbs have few side effects and abundant sources,offering new ideas and approaches for the prevention and treatment for hypertrophic scar.OBJECTIVE:To explore the potential molecular mechanism of plant-derivedβ-sitosterol on hypertrophic scar fibroblasts by network pharmacology and molecular docking techniques and to initially verify it by cytological experiments.METHODS:Through the network pharmacology,the relevant database and software were used to screen the drug targets ofβ-sitosterol and obtain the hypertrophic scar-related disease targets.The potential(intersection)targets ofβ-sitosterol on hypertrophic scar were obtained.Cytoscape software and STRING database were used to construct the“drug-target-disease”network and protein-protein interaction network,and screen out the core targets in the protein-protein interaction network.Gene ontology(GO)biological function and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses of intersection targets were conducted through the DAVID database,and the signaling pathways and core target genes closely related to the intersection targets were further identified through literature analysis.AutoDock software was used to perform the molecular docking ofβ-sitosterol and core target proteins.In vitro cellular assays were used to verify the effects ofβ-sitosterol on proliferation,apoptosis,cell cycle distribution and mRNA expression of core target genes in human hypertrophic scar fibroblasts.RESULTS AND CONCLUSION:There were 56 intersection targets ofβ-sitosterol and hypertrophic scar and 10 core targets were identified in the protein-protein interaction network,including tyrosine kinase,mitogen-activated protein kinase 3(MAPK3),cysteine protease 3(CASP3),apolipoprotein E,estrogen receptor 1,sterol regulatory element-binding transcription factor 1,peroxisome proliferator-activated receptor alpha,C-reactive prot

关 键 词：增生性瘢痕 成纤维细胞 Β-谷甾醇 细胞凋亡 MAPK3(ERK1-MAPK) 肿瘤坏死因子 CASP3 P53 网络药理学 细胞周期 

分 类 号：R452[医药卫生—治疗学] R363[医药卫生—临床医学] R364