Human Neutrophil Elastase Induced Goblet Cells Overexpression In Chronic Rhinosinusitis Through Tumour Necrosis Factor-αconverting Enzyme

Objectives： In order to provide evidence of HNE induced goblet cellsoverexpression in CRS by TACE and to further understanding the pathogenesis ofCRS,we detected the expression of human neutrophil elastase (HNE), tumor necrosisfactor-α converting enzyme (TACE) and MUC5AC in CRS patients，and researchedthe effect of HNE induce MUC5AC production by TACE in vitro cell cultureexperiments.Methods：We take the sinus mucosa of20CRS with polyps patients and20CRSwithout polyps patients as two experimental groups,and take10normol sinonasalmucosa cases as control group.We used HE staining and PAS staining to observe thepathological changes of each sample,detected the expression of HNE, TACE andMUC5AC in the sinonasal mucosa by using immunohistochemical methods, andapplied the fluorescence quantitative PCR method to detect the expression of HNE,TACE and MUC5AC mRNA in CRS. In addition,we developed a primary culturesystem of human nasal mucosa epithelial cell in vitro, and usedimmunocytochemistry methods to indentificated the cell source of the nasal mucosalepithelium cells. After the cells had stably passaged,we gave them HNE or tumornecrosis factor-converting enzyme inhibitor-1(TAPI-1)as stimulation or interventionfactors,then detected the expression of MUC5ACmRNA in cell by using fluorescentquantitative PCR method.Results：The HE staining showed that in the sinus mucosa with or withoutpolyps CRS patients,the main pathological features is the hyperplasia of goblet cell,inflammatory cell and submucosal gland.In the sinus mucosa of the two experimentalgroups,we can see they have strong expression in PAS staining,while the controlgroup only have a weak positive expression.The immunohistochemistry demonstratedthat the expression of MUC5AC, TACE and HNE in the sinus mucosa of with orwithout CRS patients were all higher than in the control patients, and the differencehave statistically significant (P<0.05), moreover,the expression have no difference (P>0.05) between the two CRS groups. The results of the fluorescence QuantitativePCR showed that, compared with the control group,the expression of MUC5AC,TACE and HNE mRNA in the two CRS groups were increased, and the differenceswere statistically significant(P <0.05), but there have no significant differencebetween the two CRS groups (P>0.05). After the nasal epithelial cells weresuccessfully cultured, we confirmed they were epithelial cells byimmunocytochemistry with cytokeratin AE1antibody.By using the the fluorescenceQuantitative PCR,we found that the expression of MUC5ACmRNA in cells that havegiven HNE was higher than that in untreated cells, and the difference wasstatistically significant (P <0.05); while the cells pretreated with tumor necrosisfactor-converting enzyme inhibitor TAPI-1and the cells simple treated with TAPI-1,there MUC5ACmRNA expressions were significantly reduced compared with theexpressions of the untreated cells and the HNE stimulated cells,and the differencehave statistically significant (P <0.01).Conclusion：The expressions of MUC5AC, TACE and HNE were all increasedin with or without CRS, and HNE can induce MUC5AC production by TACE in thatdemonstrate HNE-TACE signal path has an important role in the process of the gobletcells hyperplasia in CRS.