Protective effect of bone marrow mesenchymal stem cell-derived exosomes on HK-2 cells with hypoxia-re-oxygenation injury through regulating Beclin1 expression
Objective To investigate the protective effect and molecular mechanism of bone marrow mesenchymal stem cells(BMSCs)-derived exosomes against hypoxia-reoxygenation(H/R)injury in human tubular epithelial cells(HK-2).Methods BMSCs Exo were extracted using an extracellular vesicle kit,and their morphology was ob-served under transmission electron microscopy.Western blot was used to detect specific marker proteins on the sur-face of the extracellular vesicles.PHK67 green fluorescence was used to label the extracellular vesicles and observe the uptake phenomenon of HK-2 cells.HK-2 cells were divided into control group(NC),normal group+exosome gro-up(Exo),H/R induction group(H/R),and H/R+exosome group(H/R+Exo).CCK-8 assay was used to detect cell vi-ability;flow cytometry was used to detect cell apoptosis rate;immunofluorescence staining was used to detect reactive oxygen species(ROS)level;oxidative stress-related detection kits were used to determine MDA,GSH content;quan-titative real time polymerase chain reaction(qRT-PCR),Western blot,immunofluorescence staining were used to de-tect the expression level of Beclin1 in the HK-2 cells.Results BMSCs Exo was successfully obtained and could be taken up by HK-2 cells.Compared with NC group,cell viability and GSH level in H/R group significantly decreased,while cell apoptosis,ROS,MDA significantly improved(all P＜0.05).Compared with H/R group,cell viability,GSH level group were significantly improved and cell apoptosis,ROS,MDA were significantly decreased in H/R+E xo group(all P＜0.05).Compared with NC group,the expression level of Beclin1 in H/R group was significantly im-proved,after treatment with exosomes in H/R group,the expression level of Beclin1 significantly decreased in H/R+Exo group(all P＜0.05).Conclusions BMSCs-Exo can alleviate H/R-induced injury of HK-2 cells by downregu-lating the expression of Beclin1.
NETL
NSTL
万方数据
