Effect of adipose derived stem cell exosomes on fibrogenesis of human endometriosis cells
Objective To investigate the effects of adipose-derived stem cell-derived exosomes(ADSCs-Exos)on the proliferation and fibrosis-related factor expression of endometriosis cells(12Z).Methods The ADSCs-Exos were extracted by ultracentrifugation,identified by transmission electron microscopy,nanoparticle tracking analysis(NTA),and Western blotting.The uptake of exosomes by 12Z cells was observed by PKH26 labeling exosome method.The effects of ADSCs-Exos(12.5,25.0,50.0 μg·mL-1)on the proliferation of 12Z cells were detected by CCK-8 assay.The 12Z cells were divided into four groups:control group,model group,and ADSCs-Exos(12.5,25.0,50.0 μg·mL-1)group.Except for the control group,the other groups were induced to fibrosis by TGF-β1(5 ng·mL-1)for 24 h,and the expression of fibrosis-related gene mRNA was detected by real-time quantitative PCR(qRT-PCR)after drug treatment.Results The ADSCs-Exos extracted showed round particles that were clearly visible.The average particle size of the total extract was 122.6 nm,and the concentration was 1.8 × 106 particles·mL-1.CD9,CD63,and TSG101 were expressed in ADSCs-Exos,and Calnexin protein was not expressed,consistent with the characteristics of exosomes.12Z cells successfully internalized and phagocytosed ADSCs-Exos.The CCK-8 assay results showed that compared with the control group,the relative survival rate of 12Z cells was significantly lower after 24 h of treatment with 25.0 μg·mL-1ADSCs-Exos(P＜0.01).The qRT-qPCR results showed that compared with the control group,the expression levels of CTGF,COL1A1,and α-SMA mRNA in the model group were significantly incresed(P＜0.001).Compared with the model group,the expression levels of CTGF,COL1A1,and α-SMA mRNA in the 12.5,and 25.0 μg·mL-1 ADSCs-Exos groups were lower(P＜0.05,0.01,0.001),and the expression level of α-SMA mRNA in the 50.0 μg·mL-1 ADSCs-Exos group was significantly lower(P＜0.001).Conclusion Adipose mesenchymal stem cell exosomes can be swallowed by endometriosis cells,which can further inhibit the proliferation of endometriosis cells and reduce the expression level of cell fibrosis-related factor genes,thus playing a role in the treatment of endometriosis.
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