Mechanism of MUC5AC secretion induced by lipid-associated membrane protein of Mycoplasma pneumoniae in human bronchial epithelial cells
Objective To investigate the mechanism of mucin 5AC(MUC5AC)expression stimulated by lipid-associated membrane protein(LAMPs)of Mycoplasma pneumoniae(MP)in human bronchial epithelial cells(HBEC).Methods MP was cultured in large quantities,LAMPs were extracted,and Western blot was used to detect the MUC5AC protein level of LAMPs induced HBEC.Specific siRNA was used to silence and overexpress interleukin 17(1L-17)in HBEC cells,respectively.Blank control group,LAMPs group,LAMPs+si-IL17 group,IL-17 group,and LAMPs+IL-17 group were set up.Western blot was used to analyze the effect of LAMPs on nuclear factor-κB(NF-κB)-related signaling molecules(NF-κB p65,p-NF-κB p65,IKK-β)and MUC5AC protein expression.After HBEC cells were pretreated with NF-κB inhibitors,Western blot was used to detect the secretion level of MUC5AC after LAMPs stimulation.Results Compared with the control group,LAMPs could stimulate a significant increase in the expression of MUC5AC protein(1.68±0.31)in HBEC cells,the difference was statistically significant(t=6.38,P＜0.05).Compared with the LAMPs group,the protein expression levels of NF-κB p65,p-NF-κB p65,IKK-β and MUC5AC in the LAMPs+si-IL17 group(respectively 1.12±0.37,1.67±0.27,7.53±0.31,1.26±0.17)were significantly reduced,the differences were statistically significant(all P＜0.01).After LAMPs+overexpression of IL-17,the protein expression levels of NF-κB signaling molecules NF-κB p65,p-NF-κB p65,IKK-β and MUC5AC(respectively 1.87±0.19,3.07±0.33,3.94±0.22,2.93±0.15)were significantly increased compared with the control group,IL-17 group,and LAMPs group,the differences were statistically significant(F=6.72,12.84,21.19,11.31;all P＜0.05).After treatment with NF-κB inhibitor,the expression of MUC5AC(1.21±0.10)in LAMPs-induced cells decreased significantly compared with before treatment(1.59±0.12),the difference was statistically significant(t=2.5 1,P=0.012).Conclusion MP-LAMPs stimulated the expression of MUC5AC in HBEC cells by inducing the NF-κB signaling pathway with IL-17.
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