Exosomes of Placental MSCs Inhibit the Progression of Pulmonary Fibrosis in Silicosis by Regulating the NF-κB Signaling Pathway
Objective To explore the mechanism of human placental mesenchymal stem cells exosomes(hPMSCs-Exo)on silica(SiO2)-induced pulmonary fibrosis in mouse alveolar macrophages(MH-S).Methods Exosomes were extracted by differential ultracentrifugation Western blot,transmission electron microscopy,and nanoparticle size tracking were used to identify the exosomes.MTT assay was used to detect the effect of different concentrations of SiO2 on the viability of MH-S proliferating cells at 24 hours,combined with RT-qPCR to detect the expression of collagen typeⅠ(COL-Ⅰ)and α-smooth muscle(α-SMA)in MH-S cells,to select the optimal stimulating mass concentration of SiO2 for the subsequent experiments.MH-S cells were divided into blank control group,SiO2 group,SiO2+DMSO group,SiO2+PDTC group,SiO2+DMSO+hPMSCs group,SiO2+PDTC+hPMSCs group,SiO2+DMSO+hPMSCs-Exo group,and SiO2+PDTC+hPMSCs-Exo group.RT-q PCR was used to detect the relative transcript levels of cellular inflammatory factors IL-1β,IL-6,and TNF-α.Western blot was used to detect the NF-κB signaling pathway and apoptosis of each group.Results After SiO2 stimulation of MH-S cells,the expression of lung fibrosis-related genes COL-Ⅰ and α-SMA increased,the expression of inflammatory factors TNF-α,IL-1β and IL-6 increased.The NF-κB pathway was activated,and the expression of its related proteins and apoptotic factors were up-regulated,which contributed to the development of fibrosis(P all＜0.01).After the addition of the inhibitor PDTC,which inhibits the NF-κB signaling pathway,the NF-κB pathway was inhibited,the expression of its related proteins were reduced.The expression of inflammatory factors TNF-α,IL-1β,IL-6 were reduced,the expression of pulmonary fibrosis-related genes COL-Ⅰ,α-SMA were attenuated,the apoptosis rate was reduced,and the development of pulmonary fibrosis was inhibited(P all＜0.05).After inhibition of the NF-κB pathway in transplanted hPMSCs cells and hPMSCs-Exo,the expression of pulmonary fibrosis-related genes were significantly reduced,the release of inflammatory factors were decreased,apoptosis was inhibited,and the development of PF was effectively intervened(P all＜0.05).Conclusion hPMSCs-Exo may inhibit the progression of PF in silicosis by regulating the NF-κB signaling pathway.
human placental mesenchymal stem cells exosomespulmonary fibrosisNF-κB signaling pathwayapoptosis
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