miR-455-5p Attenuates Inflammation of Airway Epithelial Cells Infected by Respiratory Syncytial Virus by Inhibiting SOCS3
Objective:To investigate the effect of miR-455-5p on the inflammatory response of airway epithelial cells infected with respiratory syncytial virus(RSV)and its mechanism.Methods:The level of serum miR-455-5p was measured by qRT-PCR in 30 health children(health group),mild group(n=41)and severe group(n=31)children with RSV infection.16HBE cells were divided into Control group,NC-agomir group,miR-455-5p-agomir group,NC-antagomir group and miR-455-5p-antagomir group.16HBE cells were transfected by Lipofectamine 3000 and cultured for 48 h.Cell proliferation of 16HBE cells were detected by CCK-8 method and EdU method,and apoptosis was detected by TUNEL method.The levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-6 and IL-8 in the supernatant of 16HBE cells were detected by ELISA method.The mRNA levels of miR-455-5p and suppressor of cytokine signaling-3(SOCS3)were detected by qRT-PCR.The protein levels of SOCS3,interferon-α(IFN-α),signal transducer and activator of transcription(STAT)1,STAT2,p-STAT1 and p-STAT2 were detected by Western blot.16HBE cells were divided into Blank group,NC group(cells transfected with NC-agomir),RSV+NC-agomir group and RSV+miR-455-5p-agomir group.16HBE cells of RSV+NC-agomir group and RSV+miR-455-5p-agomir group were infected with RSV virus solution,and Blank group and NC group 16HBE cells were cultured nor-mally.Then,cell proliferation,apoptosis,inflammatory factors,miR-455-5p and SOCS3 mRNA levels,SOCS3,IFN-α,STAT1,STAT2,p-STAT1 and p-STAT2 protein levels were detected according to the above methods.Results:Compared with the health group,the level of serum miR-455-5p in mild group and severe group was lower than that in health group(P＜0.05).The level of serum miR-455-5p in se-vere group was lower than that in mild group(P＜0.05).Compared with Control group and NC-agomir group,miR-455-5p level,relative cell viability and EdU positive rate in miR-455-5p-agomir group increased(P＜0.05),TUNEL positive rate decreased(P＜0.05),TNF-α,IL-6 and IL-8 levels in supernatant decreased(P＜0.05),SOCS3 mRNA and protein levels decreased(P＜0.05),IFN-α protein,STAT1 and STAT2 phosphorylation increased(P＜0.05).Compared with Control group and NC-antagomir group,miR-455-5p level,relative cell via-bility and EdU positive rate in miR-455-5p-antagomir group decreased(P＜0.05),TUNEL positive rate increased(P＜0.05),TNF-α,IL-6 and IL-8 levels in supernatant increased(P＜0.05),SOCS3 mRNA and protein levels increased(P＜0.05),IFN-α protein,STAT1 and STAT2 phosphorylation decreased(P＜0.05).Compared with Blank group and NC group,miR-455-5p level,relative cell viability and E-dU positive rate in RSV+NC-agomir group decreased(P＜0.05),TUNEL positive rate increased(P＜0.05),TNF-α,IL-6 and IL-8 levels in supernatant increased(P＜0.05),SOCS3 mRNA and protein levels increased(P＜0.05),IFN-α protein,STAT1 and STAT2 phosphoryla-tion decreased(P＜0.05).Compared with RSV+NC-agomir group,miR-455-5p level,relative cell viability and EdU positive rate in RSV+miR-455-5p-agomir group increased(P＜0.05),TUNEL positive rate decreased(P＜0.05),TNF-α,IL-6 and IL-8 levels in super-natant decreased(P＜0.05),SOCS3 mRNA and protein levels decreased(P＜0.05),IFN-α protein,STAT1 and STAT2 phosphorylation levels increased(P＜0.05).Conclusion:miR-455-5p is down-regulated in the serum of RSV-infected children,and up-regulated miR-455-5p activates IFN-α-mediated antiviral response in RSV-infected 16HBE cells by inhibiting SOCS3 transcription and expression.
miR-455-5pCytokine signal transduction inhibitor 3Respiratory syncytial virusAirway epithelial cellsInflammatory response
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