海南医科大学学报2025，Vol.31Issue(19)：1458-1468,11.DOI:10.13210/j.cnki.jhmu.20241121.004
基于WGCNA分析维甲酸受体在hBMSCs成脂分化中的调控网络
Exploring regulatory network of retinoic acid receptors in adipogenic differentiation of hBMSCs based on WGCNA
摘要
Abstract
Objective:To investigate the regulatory networks of retinoic acid receptors(RARs)in the adipogenic differentia-tion of hBMSCs using weighted gene co-expression network analysis(WGCNA)and provide a basis for elucidating the molecular mechanisms of retinoic acid involved in osteoporosis.Methods:EBSeq R package was employed to analyze the differentially ex-pressed genes of hBMSCs and the samples at five time points of 0,7,14,21,and 28 days during adipogenic induction,the WGC-NA R package was used to identify the gene modules including the retinoic acid receptors related genes and construct the co-expression networks during adipogenic differentiation of hBMSCs.Cytoscape software was used to visualize the regulatory net-works.Oil Red O staining was used to detect the effects of all-trans retinoic acid(ATRA)in adipogenic phenotype of hBMSCs,qPCR was used to detect the expression of target genes.RARB shRNA lentiviral particles were used to knock down the expres-sion of the RARB gene,and the gene expression levels of the focal adhesion pathway and adipogenic marker genes were detected.Results:The genes associated with RARs(RARB,RARG,CRABP2 and RARRES1)were located in the gene module which is the most relevant to hBMSCs group,which were significantly down-regulated during adipogenic differentiation,and were close-ly related to signal pathways including focal adhesion,PI3K-Akt,ECM-receptor interaction,regulation of action cytoskeleton,TNF,Rap1,Jak-STAT,Osteoclast differentiation and MAPK etc.RXRA was located in the gene module most relevant to 7-day adipogenic induction group.It was closely related to some classic pathways(PI3K-Akt,HIF-1,Estrogen,JAK-STAT etc.)and metabolism pathways(fatty acid degradation,steroid biosynthesis,tyrosine metabolism,beta-alanine metabolism etc.).ATRA up-regulated the expression of focal adhesion pathway genes such as ITGA4,PXN,CCND1 and RAC2,and downregulated the ex-pression of adipogenic marker genes PPARG and FABP4,thus inhibiting adipogenic differentiation.After the RARB level was significantly reduced in the RARB shRNA treatment group compared to the control group,the expression of focal adhesion path-way genes was downregulated while the expression of adipogenic marker genes was upregulated.Conclusion:RARs and retinoid x receptors(RXRs)play different regulatory roles in the adipogenic differentiation of hBMSCs.RARs may be negative regulators in adipogenic differentiation,inhibiting the initiation of adipogenesis;while RXRs may be positive regulators in adipogenesis by mod-ulating cell metabolism to facilitate the development of differentiation.The inhibition of RARs on adipogenesis may be related to the focal adhesion pathway by RARβ receptor.关键词
维甲酸受体/权重基因共表达网络分析/人骨髓间充质干细胞/成脂分化/黏着斑通路Key words
Retinoic acid receptor/WGCNA/hBMSCs/Adipogenic differentiation/Focal adhesion pathway分类
医药卫生引用本文复制引用
陈思羽,彭嘉欣,农诗蕾,陈依凡,曹俊..基于WGCNA分析维甲酸受体在hBMSCs成脂分化中的调控网络[J].海南医科大学学报,2025,31(19):1458-1468,11.基金项目
This study was supported by the National Natural Science Foundation of China(81860166) 国家自然科学基金资助项目(81860166) （81860166）
