医学分子生物学杂志2025，Vol.22Issue(2)：139-145,7.DOI:10.3870/j.issn.1672-8009.2025.02.006
外泌体miR-24作为海绵体增强肾小管上皮表达S100 A8促进线粒体损伤和肾结石的致病机制研究
miR-24 Acts as Spongy of S100A8 in Kidney Tubular Epithelium Exo-somes to Enhance Mitochondrial Damage and Promote Kidney Stone Disease
摘要
Abstract
Objective To investigate theeffect of miR-24 and S100A8 in exosomes of an in vitro kidney stone cell model on kidney stone disease and the underlying mechanism.Methods Hu-man proximal tubular epithelial HKC-8 cells were used to establish a kidney stone cell model in vitro.HKC-8 cells were divided into four groups:the control group,the model group,the miR-24 mimic+model group(miR-24 mimic was transfected into the cells of the model group),and the mimic NC+model group(mimic NC was transfected into the cells of the model group).Bioinfor-matics analysis was performed to analyze the relationship of miR-24 and S100A8 3′-UTR by using TargetScan v7.2.Dual luciferase reporter assay was used to verify the direct interactions between miR-24 and S100A8 3′-UTR.Exosomes were extracted.The levels of miR-24 in exosomes was deter-mined by qPCR.Western blotting was used to determine the protein expression levels of CD9,CD63 and S100A8 in exosomes,and the protein expression levels of PINK1,Parkin and Cytochrome C(Cyt-C)in cells.The levels of inflammatory cytokines TNF-α,IL-1β and IL-6 in cell supernatants and malondialdehyde(MDA)in cells were determined by ELISA.The levels of reactive oxygen species(ROS)were determined by DCFH-DA fluorescent dye method.Results Dual luciferase reporter assays had proved a direct interactions between miR-24 and S100A8 3′-UTR.miR-24 was down-regulated and S100A8 was up-regulated in the exosomes of the Model group(P<0.05),the expression levels of mitophagy markers of PINK1,Parkin and Cytochrome C were up-regulated(P<0.05),the intracellular ROS and MDA levels were up-regulated(P<0.05),TNF-α,IL-1β and IL-6 were up-regulated in cell supernatants in the Model group(P<0.05)when compared with those in the Control group.The above indexes were all partially reversed in the miR-24 mimic+Mod-el group(P<0.05),while there had been no significant difference in the mimic NC+Model group(P>0.05)when compared with those in the Model group.Conclusion MiR-24 was down-regula-ted and as"spongy"of S100A8 in exosomes of kidney stone cell model to up-regulate S100A8 ex-pression,enhance mitochondrial damage and promote kidney stone disease.关键词
肾结石病/草酸钙结晶/肾小管上皮细胞/线粒体自噬/氧化应激/炎症Key words
kidney stone disease/calcium oxalate crystals/renal tubular epithelial cells/mitochondrial autophagy/oxidative stress/inflammation分类
临床医学引用本文复制引用
迪亚尔·地里木拉提,摆俊博,刘开放,娜菲莎·吐尔地,李佳..外泌体miR-24作为海绵体增强肾小管上皮表达S100 A8促进线粒体损伤和肾结石的致病机制研究[J].医学分子生物学杂志,2025,22(2):139-145,7.基金项目
新疆医科大学青年科研起航专项基金(No.2022YFY-QKQN-08) This work was supported by a grant from the Youth Scientific Research Initiation Special Fund Project of Xinjiang Medical University(No.2022YFY-QKQN-08) （No.2022YFY-QKQN-08）
