基于转录组学揭示GLP-1受体激动剂对肾小球系膜细胞的保护机制  

Transcriptomics-Based Exploration on the Protective Mechanism of GLP-1 Receptor Agonist on Glomerular Mesangial Cells

作　　者：王燕萍[1] 王丽静[1] 齐利琴 高若男 刘礼斌[1] 王林曦[1] WANG Yanping;WANG Lijing;QI Liqin;GAO Ruonan;LIU Libin;WANG Linxi(Department of Endocrinology and Metabolism,Fujian Institute of Endocrinology,Fujian Medical University Union Hospital,Fuzhou 350001,China)

机构地区：[1]福建医科大学附属协和医院内分泌代谢科,福建省内分泌研究所,福州350001

出　　处：《福建医科大学学报》2024年第5期298-306,共9页Journal of Fujian Medical University

基　　金：福建省自然科学基金项目(2019J01150,2021J01757)。

摘　　要：目的探讨胰高血糖素样肽-1受体激动剂(GLP-1RA)对糖尿病肾损伤保护的相关机制。方法选用小鼠肾小球系膜细胞(MCs)为体外研究对象,并分为对照组(C组)、高糖高脂组(GP组)和GLP-1RA exendin-4处理组(EX-GP组)。采用CCK-8检测细胞存活率;采用转录组测序(RNA-seq)对细胞进行转录组测序;采用qRT-PCR测定磷酸果糖激酶(Pfk)、柠檬酸合酶(Cs)、α-酮戊二酸脱氢酶(Ogdh)、微管相关蛋白轻链3(LC3)、重组人自噬效应蛋白(Beclin-1)、线粒体DNA(mtDNA)、沉默信息调节因子1(Sirt-1)和过氧化物酶体增殖物激活受体辅助活化因子1α(Pgc-1α)的基因表达。结果在高糖高脂诱导的肾小球MCs损伤模型中,GP组MCs存活率降低,EX-GP组存活率恢复。RNA-seq结果显示,3组间的基因表达有明显差异。通过京都基因与基因组百科全书(KEGG)富集分析,基因差异表达通路涉及细胞脂肪酸代谢过程、mTOR信号通路、自噬等。GP组抑制了Pfk表达,EX-GP组可部分恢复其表达(P<0.05)。exendin-4处理后,mtDNA、LC3和Beclin-1基因表达均增加(P<0.05)。与GP组比较,EX-GP组的Sirt-1和Pgc-1α基因表达升高(P<0.05)。结论RNA-seq及能量代谢相关基因研究提示,GLP-1RA exendin-4改善高糖高脂诱导的MCs损伤机制可能涉及能量代谢相关通路以及Sirt-1、Pgc-1α和自噬、糖酵解基因的表达。Objective To explore the protective mechanism of glucagon-like peptide-1 receptor agonist(GLP-1RA)against diabetic kidney injury.Methods In this study,mouse glomerular mesangial cells(MCs)were selected for in vitro study,and the glomerular MCs were divided into the control group(Group C),the high-glucose and high-fat treatment group(Group GP)and the GLP-1RA exendin-4 treatment group(Group EX-GP).The cell viability was measured by CCK-8;the transcriptome of the cells was sequenced by RNA-seq;the gene expression of phosphofructokinase(Pfk),citrate synthase(Cs),α-ketoglutarate dehydrogenase(Ogdh),microtubule-associated protein light chain 3(LC3),recombinant gene expression of human autophagy effector protein(Beclin-1),mitochondrial DNA(mtDNA),silencing information regulator 1(Sirt-1)and peroxisome proliferator-activated receptor-assisted activator 1α(Pgc-1α)were determined by qRT-PCR.Results In the high glucose and high fat-induced glomerular MCs injury model,the survival rate of MCs was reduced in the GP group and restored in the EX-GP group.By using RNA-seq,there were significant gene expression differences among Group C,GP and EX-GP.By KEGG enrichment analysis,the gene differential expression pathways involved cellular fatty acid metabolism,mTOR signaling pathway,autophagy and so on.The expression of Pfk gene was inhibited in GP group,but its expression could partially restore in the EX-GP group(P<0.05).After exendin-4 treatment,mtDNA gene expression was increased(P<0.05),and the expressions of autophagy related genes LC3 and Beclin-1 were significantly increased(P<0.05).The expression of Sirt-1 and Pgc-1αgenes in EX-GP group was increased compared with GP group(P<0.05).Conclusion RNA-seq and energy metabolism-related gene studies suggested that the mechanism of GLP-1RA exendin-4 amelioration of high-glucose and high-fat-induced injury in MCs may involve the energy[JP1]metabolism-related signaling pathway and the expression of Sirt-1,Pgc-1α,autophagy and glycolysis genes.

关 键 词：糖尿病肾病 胰高血糖素样肽-1受体激动剂 肾小球系膜细胞 能量代谢 

分 类 号：R692.9[医药卫生—泌尿科学] R587.2[医药卫生—外科学]