Exosomes derived from mesenchymal stem cells inhibit the proliferation,invasion and inflammation of rheumatoid arthritis fibroblast-like synoviocytes through microRNA-125b-5p
Objective To investigate the biological effects and mechanisms of mesenchymal stem cell-extracellular vesicles(MSC-EVs)on rheumatoid arthritis(RA)fibroblast-like synoviocytes(MH7A).Methods MH7A cell line was divided into control group and MSC-EVs group.MSC-EVs were collected from the culture supernatant of MSC by differential centrifugation.The control group was cultured with rou-tine methods,and MH7A cells were incubated with MSC-EVs for 24 h in the MSC-EVs group.After the two groups of cells reached the sample collection conditions,the immunofluorescence experiment,Tran-swell experiment,enzyme-linked immunosorbent assay(ELISA),and quantitative real-time polymerase chain reaction(qRT-PCR)were used to observe the effect of MSC-EVs on the biological behavior of MH7A cells and its molecular mechanism.The independent sample t test was used for statistical analysis.Results The results of nanoparticle tracking analysis and transmission electron microscopy showed that MSC-EVs were successfully extracted by differential centrifugation,and then immunofluorescence confirmed that MSC-EVs could be uptaken by MH7A cells.The results of cell proliferation showed that the gray value of proliferation cell nuclear antigen(Ki-67)fluorescence in the MSC-EVs group(4.01±1.01)was signifi-cantly lower than that in the control group(11.67±1.53)(4=-7.273,P<0.01).Transwell results showed that the invasion ability of MSC-EVs group(70.00±5.29)was significantly lower than that of the control group(110.67±6.11)(t=-8.714,P<0.01).ELISA results showed that the levels of inflam-matory factors in the MSC-EVs group were significantly lower than those in the control group[tumor necro-sis factor-α(TNF-α):t=-5.345,P<0.01;interleukin(IL)-1β:t=-3.742,P<0.05;IL-6:t=-7.579,P<0.01;IL-8:t=-6.390,P<0.01].The results of qRT-PCR showed that the expression level of microRNA(miR)-125b-5p in the MSC-EVs group(14.83±2.48)was significantly higher than that in the control group(1.01±0.15)(t=-8.714,P<0.01).Conclusion MSC-EVs could inhibit the prolifera-tion,invasion and inflammation of MH7A cells by up-regulating the expression of miR-125b-5p.