西南医科大学学报2026，Vol.49Issue(3)：335-342,8.DOI:10.3969/j.issn.2096-3351.2026.03.012
AKI血清预处理BMSCs外泌体减轻HK-2细胞缺氧/复氧损伤的体外研究
In Vitro Study on Exosomes Derived from BMSCs Preconditioned with AKI Serum Alleviating Hypoxia/Reoxygenation Injury in HK-2 Cells
摘要
Abstract
Objective The aim of this study was to investigate the protective effect of exosomes derived from acute kidney injury(AKI)serum-preconditioned bone marrow mesenchymal stem cells(AKI-Exos)on hypoxia/reoxygenation(H/R)-induced injury in Human Kidney-2(HK-2)cells,and to determine whether this protection is mediated through inhibition of pyroptosis.Methods HK-2 cells were treated with different concentrations of cobalt chloride(CoCl₂).The CCK-8 assay was employed to screen for the appropri-ate concentration and duration to establish an in vitro hypoxia/reoxygenation(H/R)injury model.The experiment included the follow-ing groups:a blank control group,an H/R model group,an H/R+normal exosomes(Nor-Exo)group,and an H/R+AKI serum-preconditioned exosomes(AKI-Exo)group.Bone marrow mesenchymal stem cells were preconditioned with either normal fetal bovine serum or rat AKI serum,and Nor-Exos and AKI-Exos were isolated by ultrafiltration.The morphology and phenotype of exosomes were characterized by transmission electron microscopy,nanoparticle tracking analysis,and Western blotting for exosomal markers Alix,CD63 and TSG101.Uptake of exosomes by HK-2 cells was observed using fluorescent labeling.ELISA was employed to detect IL-1β and IL-18 levels in the cell culture supernatant of each group.Western blotting was used to assess the expression of pyroptosis-related proteins NLRP3,GSDMD,and IL-1β.Results Nor-Exos and AKI-Exos were successfully prepared and identified.Both exhibited typical bilayer membrane vesicular morphology,with particle sizes mainly within the exosomal range,expressed exosomal markers Alix,CD63,and TSG101,and could be internalized by HK-2 cells.Compared with the blank control group,H/R treatment markedly reduced HK-2 cell viability,significantly increased IL-1β and IL-18 levels in the supernatant,and upregulated the expression of pyroptosis-related proteins NLRP3,GSDMD,and IL-1β.Compared with the H/R group,both the H/R+Nor-Exo group and H/R+AKI-Exo group showed restored cell viability,reduced IL-1β and IL-18 levels,and decreased expression of NLRP3,GSDMD,and IL-1β to varying degrees,with a more pronounced improvement observed in the AKI-Exo group.Conclusion Exosomes derived from AKI serum-preconditioned bone marrow mesenchymal stem cells can attenuate H/R-induced injury and inflammatory responses in HK-2 cells by suppressing pyroptosis.These findings suggest that AKI-Exos may represent a potential therapeutic strategy for ischemia-reperfusion-related acute kidney injury.关键词
间充质干细胞/外泌体/缺氧/复氧损伤/细胞焦亡/肾小管上皮细胞/急性肾损伤Key words
Mesenchymal stem cells/Exosomes/Hypoxia/reoxygenation injury/Pyroptosis/Renal tubular epithelial cells/Acute kidney injury分类
医药卫生引用本文复制引用
刘浪,高一丹,陈冰雪,叶青青,雷碧,廖远洋,欧阳亚玲,刘慧..AKI血清预处理BMSCs外泌体减轻HK-2细胞缺氧/复氧损伤的体外研究[J].西南医科大学学报,2026,49(3):335-342,8.基金项目
四川省医学会科研基金(Q21089) （Q21089）
四川省医学科技创新研究会项目(HR25051) （HR25051）
西南医科大学附属医院科研基金(21071) （21071）
西南医科大学科研基金(2022QN112) （2022QN112）
