黄芪治疗局灶节段性肾小球硬化的分子作用机制及miRNA-mRNA调控网络构建  

Molecular Mechanism of Astragalus on Focal Segmental Glomerulosclerosis and mi RNA-m RNA Regulatory Networks

作　　者：程馨缘 杜根发 韩鹏勋[1] 王梦华 孙惠力[1] 易铁钢[1] Cheng Xinyuan;Du Genfa;Han Pengxun;Wang Menghud;Sun Huili;Yi Tiegang(The Fourth Clinical Medical College,Guangzhou University of Chinese Medicine,Shenzhen 518033,China;Guangzhou University of Chinese Medicine,Guangzhou 510405,China;Affiliated Hospital of Traditional Chinese Medicine,Guangzhou Medical University,Guangzhou 510180,China)

机构地区：[1]广州中医药大学第四临床医学院,深圳市中医院,广东深圳518033 [2]广州中医药大学,广州510405 [3]广州医科大学附属中医医院,广州510180

出　　处：《世界科学技术-中医药现代化》2023年第1期107-119,共13页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：国家自然科学基金委员会青年项目(82004156):从TFAM-STING通路探讨蒿甲醚改善肾小管上皮细胞重塑的分子机制,负责人:韩鹏勋;深圳市科技计划委员会基础研究项目(JCYJ20190812183603627):基于丙酮酸脱氢酶激酶调控Warburg效应探讨青蒿素衍生物抗糖尿病肾脏肥大的分子代谢基础,负责人:孙惠力。

摘　　要：目的通过网络药理学及生物信息学结合体内实验探讨黄芪治疗局灶节段性肾小球硬化的分子作用机制,并构建miRNA-mRNA调控互作网络。方法从GEO、CTD、DisGeNET和GeneCards数据库获取局灶节段性肾小球硬化疾病基因,借助TCMSP、BATMAN-TCM数据库及前期研究获得黄芪主要活性成分及相应靶点,筛选中药-疾病的交集基因为两者的共同靶点,并构建中药-成分-靶点-疾病调控网络及通路富集分析。STRING数据库构建蛋白互作网络(PPI),鉴定核心基因及其miRNAs,并进行分子对接验证。利用阿霉素建立肾小球硬化小鼠模型,活性成分黄芪甲苷干预后,Western blot检测核心靶点Akt1、p-Mapk1表达量。结果黄芪-疾病共有靶点146个,涉及槲皮素、山奈酚、甜菜碱和黄芪甲苷等主要活性成分及免疫-炎症、自噬-凋亡等相关通路,其中TNF、TLRs、PI3K-Akt及MAPK是核心信号通路。核心靶点有TNF、AKT1、JUN、IL6、IL1β、CASP3、VEGFA和MAPK1,及核心miRNAs有has-miR-203a-3p、has-miR-155-5p、has-miR-106a-5p和has-miR-34a-5p。分子对接显示核心靶点与相应成分结合活性良好;体内实验显示,黄芪甲苷上调Akt1、抑制p-MAPK1蛋白表达,改善FSGS肾损伤。结论黄芪可能主要通过槲皮素、山奈酚、甜菜碱、黄芪甲苷等主要成分经免疫-炎症和自噬-凋亡相关通路作用于局灶节段性肾小球硬化症,其中核心靶点和miRNAs在治疗过程中可能发挥重要作用。Objective Based on network pharmacology and bioinformatics combined with in vivo experiments to explore the Molecular mechanism of Astragalus on focal segmental glomerulosclerosis(FSGS),and to construct a miRNA-mRNA regulatory networks.Methods The genes of focal segment glomerulosclerosis were obtained from GEO,CTD,DisGeNET and GeneCards databases,and the main active components and corresponding targets of Astragalus were screened from TCMSP and BATMAN-TCM databases,and the overlap of Astragalus and FSGS genes were used as common targets.After that,the Astragalus-component-target-FSGS regulation network and pathway enrichment analysis were performed.Then protein interaction network(PPI) was constructed using STRING database to identify hub genes and miRNAs,and molecular docking verification was performed.The mouse model of glomerulosclerosis was established by adriamycin,After the intervention of astragaloside IV,and the expression levels of core targets Akt1 and p-Mapk1 were detected by Western blot.Results A total of 146 common targets of astragalus and FSGS were obtained,mainly involving active components of quercetin,kaempferol and astragaloside IV and the pathways of immuneinflammation and autophagy-apoptosis.TNF,TLRs,PI3K-Akt and MAPK are the core signaling pathways.The hub targets(TNF,AKT1,JUN,IL6,IL1β,CASP3,VEGFA and MAPK1) and hub miRNAs(has-miR-203a-3p,has-miR-155-5p,has-miR-106a-5p and has-miR-34a-5p)were identified.Molecular docking showed that the core target had good binding activity with corresponding compounds.In vivo experiments showed that astragaloside IV up-regulated Akt1,inhibited p-MAPK1 protein expression,and improved FSGS renal injury.Conclusion Astragalus may act on focal segmental glomerulosclerosis through quercetin,kaempferol,betaine,astragaloside IV via immune inflammation and autophagy apoptosis pathways.These core targets and miRNAs may play an important role in the process of the treatment.

关 键 词：黄芪 局灶节段性肾小球硬化 网络药理学 生物信息学 分子对接 

分 类 号：R-058[医药卫生]