Human Neutrophil Elastase Mediates MUC5AC Hypersecretion via the Tumour Necrosis Factor-α Converting Enzyme-Epidermal Growth Factor Receptor Signalling Pathway in vivo
Human Neutrophil Elastase Mediates MUC5AC Hypersecretion via the Tumour Necrosis Factor-α Converting Enzyme-Epidermal Growth Factor Receptor Signalling Pathway in vivo
复制标题
人中性粒细胞弹性蛋白酶通过体内肿瘤坏死因子-α 转换酶-表皮生长因子受体信号通路介导 MUC5AC 分泌过多
DOI：
10.1159/000509982
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发表时间：
2021-06-15
影响因子：
1.3
通讯作者：
Luo，Qing
中科院分区：
文献类型：
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作者：
Zhao，Junmei;Yang，Tian;Luo，Qing
目的探讨肿瘤坏死因子-α转化酶-表皮生长因子受体(TACE-EGFR)通路在人中性粒细胞弹性蛋白酶(HNE)诱导小鼠MUC5AC粘蛋白表达中的作用。苏木精-伊红(HE)和高碘酸-席夫(PAS)染色观察组织病理学变化。免疫组织化学方法检测TACE、EGFR和MUC5AC在鼻黏膜中的表达。用免疫印迹法检测p-EGFR、EGFR和TACE蛋白的表达，用ELISA法检测MUC5AC蛋白的表达水平。结果与对照组相比，HNE组鼻黏膜组织中可见不规则的上皮细胞、杯状细胞和粘膜下腺的增生。在鼻粘膜，PAS染色可见强阳性紫红色颗粒，TACE、EGFR、MUC5AC的mRNA和蛋白表达显著增加(p<；0.01)。HNE+TAPI-2组和HNE+AG1478组杯状细胞和粘膜下腺增生明显减少，PAS染色减弱。与HNE单独治疗组相比，HNE+TAPI-2治疗组小鼠TACE、EGFR、MUC5AC基因和蛋白以及p-EGFR蛋白的表达水平显著降低(p<；0.01)。在HNE+AG1478组小鼠，EGFR和MUC5AC的mRNA和蛋白水平以及p-EGFR蛋白的表达显著降低(p<；0.01)，而TACE的mRNA和蛋白的表达在HNE+AG1478组和HNE组之间无显著差异(p>；0.05)。结论通过建立一种新的、稳定的小鼠鼻高分泌模型，TAPI-2或AG1478可以抑制HNE诱导的MUC5AC的产生。这表明MUC5AC粘蛋白在体内的表达是由涉及HNE-TACE-EGFR信号通路的级联反应所介导的。
ObjectivesThe objective of this study is to examine the role of the tumour necrosis factor-α converting enzyme-epidermal growth factor receptor (TACE-EGFR) pathway in human neutrophil elastase (HNE)-induced MUC5AC mucin expression in mice.MethodFour groups of mice, treated with HNE alone (HNE group), HNE plus TACE inhibitor (HNE+ TAPI-2 group), HNE plus EGFR inhibitor (HNE+ AG1478 group), and untreated (control group), were used in the experiment. Histopathological changes were monitored by haematoxylin-eosin (HE) and periodic acid-Schiff (PAS) staining. TACE, EGFR, and MUC5AC expression in the nasal mucosa were determined using immunohistochemistry. The expression of p-EGFR, EGFR, and TACE protein was analysed on Western blots, and MUC5AC protein levels were assessed via ELISA. TACE, EGFR, and MUC5AC expression in the nasal mucosa were determined using real-time quantitative PCR.ResultsCompared to the control group, HE-stained tissues from the HNE group showed an irregular epithelium as well as goblet cell and submucosal glandular hyperplasia. In the nasal mucosa, strongly positive fuchsia granules were seen in PAS staining and significant increases in TACE, EGFR, MUC5AC mRNA, and protein expression were detected (p< 0.01). The HNE+ TAPI-2 and HNE+ AG1478 groups had significantly less goblet cell and submucosal gland hyperplasia as well as weaker PAS staining. Compared to mice treated with HNE alone, in HNE+ TAPI-2-treated mice, the levels of TACE, EGFR, and MUC5AC mRNA and protein as well as p-EGFR protein were significantly reduced (p< 0.01). In HNE+ AG1478-treated mice, EGFR and MUC5AC mRNA and protein levels and p-EGFR protein expression were reduced significantly (p< 0.01), but the difference in TACE mRNA and protein expression between the HNE+ AG1478 and HNE groups was not significant (p> 0.05).ConclusionUsing a newly developed, stable experimental model of nasal hypersecretion in mice, we showed that TAPI-2 or AG1478 inhibited HNE-induced MUC5AC production. This suggests that MUC5AC mucin expression in vivo is mediated by a cascade involving the HNE-TACE-EGFR signalling pathway.