HNE Induced MUC5AC Overexpression In CRSwNP Through TRAF6-Autophagy Pathway

Objective:Investigated the role of TRAF6 in the reduction of HNE-induced mucin MUC5 AC expression in CRSwNP.Methods:1、The expression of MUC5 AC,TRAF6,LC3 B,BECN1 and ATG5 in nasal mucosa of CRSwNP group and negative control group were detected by real-time quantitative PCR(q RT-PCR),immunehistochemistry(IHC)and enzyme linked immunosorbent assay(ELISA).2、Primary human nasal epithelial cells(HNECs)were isolated from normal nasal mucosa and cultured in vitro,and after HNE stimulation,the expression of MUC5 AC,TRAF6,BECN1,ATG5 and LC3 B was detected by western blotting,ELISA and q RT-PCR,HNECs were treated with HNE,Then treated with autophagy inhibitor 3-MA(3-Methyladenine),Cultured cells were analyzed using western blotting,q RT-PCR,and ELISA.3、Cultured primary HNECs were transfected with small interfering RNAs(si RNAs)to target TRAF6.Cultured cells were analyzed using western blotting,q RT-PCR,and ELISA.Using TRAF6 inhibitor acted on HNECs,Then treated with HNE,the expression trend of MUC5 AC,TRAF6,BECN1,ATG5 and LC3 B were detected by western blotting,q RT-PCR,and ELISA.Results:1、IHC,qRT-PCR and Western blot showed that the expression of TRAF6,ATG5,BECN1,ATG5 and MUC5AC in CRSwNP was up-regulated compared with normal control group(P < 0.05).2、Compared with the control group,the expression of MUC5 AC,BECN1,ATG5 and TRAF6 was up-regulated in HNE group,In autophagy inhibition group,the expression of LC3 B BECN1,ATG5 and MUC5AC was down regulated,And there was no significant difference in TRAF6 expression.(p <0.05).3、HNE-induced secretion of MUC5 AC was suppressed in normal primary HNECs by TRAF6-si RNA;After TRAF6 inhibitor action,the secretion trend of TRAF6 and MUC5AC showed the opposite trend to the normal group(p <0.05).Conclusions:HNE promotes autophagy through TRAF6,resulting in overexpression of MUC5 AC In CRSwNP.