中兽医医药杂志2025，Vol.44Issue(5)：1-11,11.DOI:10.13823/j.cnki.jtcvm.2025.048
续断-熟地黄提取物对IR-HepG2细胞糖代谢的调节及作用靶点分析
Regulation of glucose metabolism in IR-HepG2 cells by Dipsaci Radix-Rehmanniae Radix Praeparata extract and analysis of its mechanism of action
摘要
Abstract
This study aimed to investigate the mechanism of Dipsaci Radix and Rehmanniae Radix Praeparata compound extract on regulating glucose metabolism in HepG2 cells by using network pharmacology,molecular docking analysis,and cellular experiments,and provide reference material for the treatment of type 2 diabetes in dogs and cats by traditional Chinese medicine.Initially,active compounds of Dipsaci Radix and Rehmanniae Radix Praeparata along with their disease-related targets were retrieved from TCMSP,GeneCards,and OMIM databases.Subsequently,protein-protein interaction(PPI)network analysis,Gene Ontology and KEGG pathway enrichment analyses were performed,followed by molecular docking validation of binding affinity between core compounds and key targets.Finally,optimal herbal concentrations were determined via MTT assay.Insulin-resistant HepG2 cell models were established under hyperglycemic and hyperinsulinemic conditions.The regulatory effects on glucose metabolism were verified by quantifying mRNA expression of core targets and glycolytic/gluconeogenic enzymes using RT-qPCR.The result showed that 26 active compounds from Dipsaci Radix and Rehmanniae Radix Praeparata were identified,among which sylvestrosideⅢ_qt,japonine and cauloside A_qt were core constituents.A total of 402 overlapping targets were determined,among which STAT3,EGFR,HSP90AA1,AKT1,and IL-6 were core therapeutic targets for type 2 diabetes mellitus(T2DM).Cellular assays demonstrated the optimal concentration of the herbal compound extract was 10-6 g/mL,which could significantly promote cell proliferation(P<0.05).RT-qPCR results indicated the extract could significantly upregulate the mRNA expression of AKT1(P<0.01),INSR(P<0.01),GLUT4(P<0.05),AMPK(P<0.05),GCK(P<0.01)and GYS2(P<0.01),and significantly downregulate the mRNA expression of STAT3(P<0.001),EGFR(P<0.05),HSP90AA1(P<0.01),IL-6(P<0.001),GSK-3β(P<0.01),PEPCK(P<0.05)and G6Pase(P<0.001).It is concluded that Dipsaci Radix and Rehmanniae Radix Praeparata can ameliorate insulin resistance through multi-component,multi-target,and multi-pathway regulation,effectively alleviate cellular damage from insulin resistance,downregulate inflammatory responses,enhance the expression of key insulin signaling proteins,and mitigate glucose metabolism dysfunction.These findings establish a mechanistic foundation for clinical treatment of canine and feline diabetes with Dipsaci Radix and Rehmanniae Radix Praeparata.关键词
续断-熟地黄/网络药理/HepG2细胞/胰岛素抵抗/糖代谢Key words
Dipsaci Radix-Rehmanniae Radix Praeparata/network pharmacology/HepG2 cells/insulin resistance/glucose metabolism分类
农业科技引用本文复制引用
赫英辉,岳自鹏,郝淋耀,李树鹏,李玉荣,霍书英..续断-熟地黄提取物对IR-HepG2细胞糖代谢的调节及作用靶点分析[J].中兽医医药杂志,2025,44(5):1-11,11.基金项目
河北农业大学科研发展基金(JY2018005) （JY2018005）
