Lycopene Inhibits Inflammatory Airway Mucous Hypersecretion Through Epidermal Growth Factor Receptor

Objective: To research the therapeutic effect and related signalingpathways of lycopene on the pathological mucin（MUC）5ac in the mucushypersecretion airway epithelial cells model that stimulated by humanneutrophil elastase（HNE）.Methods: We divide the human bronchial epithelial cells of the lineHBE16that were cultured into seven groups，each group were treated with0,25,50,75,100,200,400umol/L lycopene.Using methyl thiazolyltetrazolium(MTT) absorption method to observe the survival of the cellsHBE16in each group,and we make sure the role of lycopene dose. Thecultured cells HBE16were divided the cells into five groups， andintervented with lycopene and epidermal growth factor receptor inhibitorAG1478:①the negative control group;②the HNE treated group at adosage of50nmol/L;③the HNE (50nmol/L) and lycopene (100umol/L)treated group;④the HNE (50nmol/L) and AG1478(5umol/L) treated group;⑤the HNE (50nmol/L),lycopene (100umol/L) and AG1478(5umol/L)treatment groups. The mRNA expression of MUC5ac and EGFR in each group were measured by Reverse transcription-PCR(RT-PCR);Measuringthe MUC5ac protein distribution with enzyme linked immunosorbentassay(ELISA);To further observe the MUC5ac protein expression of cellsintervention with lycopene, cellular immunofluorescence and laserconfocal microscope technology was provided. Using Western Blot todetect the protein expression of EGFR and p-EGFR in each group cells.Results: The cells HBE16showed a concentration-dependent.Thecells have no obvious damage when the concentration of lycopene was lessthan or equal with100umol/L.While the concentration of lycopene isgreater than100umol/L,dead cells and adherent cells graduallyincreased,MTT results showed that the best role concentration of lycopeneis100umol/L. To compared with the control group, the absorbance areavalues of MUC5ac mRNA and MUC5ac protein was significantlyincreased in HNE treatment group;we also learn that the expression ofMUC5ac protein significantly increased imaging from cellularimmunofluorescence and laser confocal microscope technology in HNEtreatment group;The expression changes of EGFR mRNA, EGFR proteinand p-EGFR protein were significantly grow in number in HNE treatmentgroup. The lycopene pretreatment group compared with HNE-treated group,The expression changes of MUC5ac mRNA and protein, EGFR mRNA andprotein, P-EGFR protein were significantly deduced;we further observethat the protein expression of MUC5AC increasingly deduced from the confocal imaging.Comparing the lycopene and AG1478pretreatment groupwith the HNE and AG1478pretreatment group, the expression changes ofMUC5ac mRNA and protein, EGFR mRNA and protein, p-EGFR proteinobviously decreased.All the results that confirmed by the statisticalmethod shows that the differences had statistical significannt(P <0.01).Conclusion: Lycopene inhibits airway mucous hypersecretion byreducing EGFR and blocking EGFR signaling pathways.At the meantime,the research shows that lycopene is feasible for the therapy of airwaymucous hypersecretion related diseases.