间充质干细胞来源外泌体减轻少突胶质细胞损伤的机制研究OA

Objective To investigate whether mesenchymal stem cell-derived exosomes(MSC-Exo)regulate ferroptosis by modulating the JAK2/STAT3 signaling pathway to alleviate hypoxia/re-oxygenation(H/R)-induced oligodendrocyte injury.Methods Oligodendrocytes were randomly divided into control,H/R,and MSC-Exo groups.Except for the control group,cells underwent H/R treatment.Western blot analysis was used to detect protein expression of oligodendrocyte markers(MAG,Olig2,MOG,MBP),exosome markers(TSG101,CD81,CD63),endoplasmic reticulum protein(calnexin),ferroptosis-related proteins(GPX4,TFR),and JAK2/STAT3 signaling pathway components(JAK2,p-JAK2,STAT3,p-STAT3).Cell viability was assessed by CCK-8 assay.Spectrophotometric methods were employed to measure caspase-3 activity,glutathione(GSH)content,Fe2+levels,and malondialdehyde(MDA)concentrations.Apoptosis and reactive oxygen species(ROS)production were analyzed by flow cytometry using Annexin-V/PI double staining.To verify the role of JAK2/STAT3 pathway in MSC-Exo-mediated inhibition of ferroptosis,a JAK2 inhibitor(JAK2i)was applied,and cells were assigned to H/R,MSC-Exo,JAK2i,and JAK2i+MSC-Exo groups.The protein expression levels of GPX4,TFR,p-JAK2,and p-STAT3,as well as cell viability,GSH,Fe2+,MDA,apoptosis,and ROS production,were determined using the aforementioned methods.Results Compared with the control group,the H/R group showed significantly decreased cell viability and GSH content,and increased ROS,MDA,Fe2+,and apoptosis(P<0.05).Protein levels of TFR,p-JAK2,and p-STAT3 were significantly upregulated,while GPX4 was downregulated(P<0.05).Compared with the H/R group,both JAK2i and MSC-Exo treatments significantly inhibited p-JAK2,p-STAT3,and TFR expression,upregulated GPX4 expression,increased cell viability and GSH content,and reduced ROS,MDA,Fe2+,and apoptosis levels(P<0.05).Conclusions MSC-Exo inhibit ferroptosis by suppressing the JAK2/STAT3 signaling pathway,thereby alleviating H/R-induced oligodendrocyte injury.