慢性鼻窦炎患者MMP-9的表达及与上皮间质转化的相关性研究

doi:10.11958/20231439

天津医药 ›› 2024, Vol. 52 ›› Issue (3): 245-249.doi: 10.11958/20231439

慢性鼻窦炎患者MMP-9的表达及与上皮间质转化的相关性研究

宋喜(), 葛益林^△(), 李殷, 宋辉, 成嘉明

长沙市第一医院耳鼻咽喉头颈外科（邮编410005）

收稿日期:2023-09-20 修回日期:2023-11-06 出版日期:2024-03-15 发布日期:2024-03-13
通讯作者: ^△E-mail：paigug@163.com
作者简介:宋喜（1981），男，主治医师，主要从事耳鼻咽喉常见病的诊断和治疗方面研究。E-mail：sx18196737@126.com
基金资助:
湖南省卫生健康委员会科研计划项目(202107011445)

Expression of MMP-9 in patients with chronic rhinosinusitis and its correlation with epithelial-mesenchymal transition

SONG Xi(), GE Yilin^△(), LI Yin, SONG Hui, CHENG Jiaming

Department of Otolaryngology, Head and Neck Surgery, Changsha First Hospital, Changsha 410005, China

Received:2023-09-20 Revised:2023-11-06 Published:2024-03-15 Online:2024-03-13
Contact: ^△E-mail: paigug@163.com

宋喜, 葛益林, 李殷, 宋辉, 成嘉明. 慢性鼻窦炎患者MMP-9的表达及与上皮间质转化的相关性研究[J]. 天津医药, 2024, 52(3): 245-249.

SONG Xi, GE Yilin, LI Yin, SONG Hui, CHENG Jiaming. Expression of MMP-9 in patients with chronic rhinosinusitis and its correlation with epithelial-mesenchymal transition[J]. Tianjin Medical Journal, 2024, 52(3): 245-249.

摘要/Abstract

摘要：

目的探讨基质金属蛋白酶（MMP）-9参与慢性鼻窦炎（CRS）上皮间质转化（EMT）的过程及机制。方法收集42例需手术治疗的CRS患者息肉样变的中鼻甲组织以及8例行鼻中隔成形术患者的中鼻甲组织，免疫组织化学染色、qPCR和Western blot检测MMP-9的表达。体外培养原代人鼻上皮细胞HNEpc，分为Control组、TGF-β1组（5 μg/L TGF-β1干预）和TGF-β1+si-MMP-9组（转染si-MMP-9和5 μg/L TGF-β1干预）。细胞免疫荧光染色检测MMP-9表达，Western blot检测TGF-β1、MMP-9和EMT相关蛋白E-钙黏蛋白（E-Cadherin）、波形蛋白（vimentin）、α-平滑肌肌动蛋白（α-SMA）表达。结果（1）CRS伴鼻息肉（CRSwNP）组患者鼻黏膜组织中MMP-9表达阳性率（54.5%，12/22）明显高于CRS不伴息肉（CRSsNP）组（25.0%，5/20）和对照组（12.8%，1/8）。CRSwNP组患者鼻黏膜中MMP-9 mRNA和蛋白相对表达水平均高于CRSsNP组和对照组（P<0.05）。（2）与Control组相比，TGF-β1组TGF-β1、MMP-9、vimentin、α-SMA表达升高，E-cadherin表达下降（P<0.05）。与TGF-β1组相比，TGF-β1+si-MMP-9组TGF-β1、MMP-9、vimentin、α-SMA表达下降，E-cadherin表达升高（P<0.05）。结论 CRS患者MMP-9表达升高，其可能通过调控EMT来参与CRS发生发展。

关键词: 慢性鼻窦炎, MMP-9, TGF-β1, 上皮间质转化, 组织重塑

Abstract:

Objective To investigate the mechanism of matrix metalloproteinase (MMP)-9 involved in epithelial mesenchymal transformation (EMT) in chronic sinusitis (CRS). Methods The expression of MMP-9 from polypoid middle turbinate tissue was detected by immunohistochemical staining qPCR and Western blot assay in 42 patients with CRS and 8 patients underwent septoplasty. Primary human nasal epithelial cells HNEpc were cultured in vitro and divided into the control group, the TGF-β1 group (5 μg/L TGF-β1 intervention) and the TGF-β1+si-MMP-9 group (transfected with si-MMP-9 and 5 μg/L TGF-β1 intervention). The expression of MMP-9 was detected by cell immunofluorescence staining. Expression levels of TGF-β1, MMP-9 and EMT-related proteins E-cadherin, vimentin and α-SMA were detected by Western blot assay. Results (1) The positive expression rate of MMP-9 was significantly higher in the nasal mucosa of CRS with nasal polyps (CRSwNP) group (54.5%, 12/22) than that of the CRS without polyps (25.0%, 5/20) group and the control group (12.8%, 1/8). The relative expression levels of MMP-9 mRNA and protein in nasal mucosa were higher in the CRSwNP group than those in the CRSsNP group and the control group (P<0.05). (2) Compared with the control group, the expressions levels of TGF-β1, MMP-9, vimentin and α-SMA were increased in the TGF-β1 group, while the expression of E-cadherin was decreased (P<0.05). Compared with the TGF-β1 group, expression levels of TGF-β1, MMP-9, vimentin and α-SMA were decreased in the TGF-β1+si-MMP-9 group, and the expression of E-cadherin was increased (P<0.05). Conclusion The expression of MMP-9 is increased in CRS patients, which may be involved in the development of CRS through the regulation of EMT.

Key words: chronic sinusitis, MMP-9, TGF-β1, epithelial mesenchymal transformation, tissue remodeling

中图分类号:

R765.4

图/表 6

图1 免疫组化染色检测3组患者鼻黏膜组织MMP-9表达（×400）

Fig.1 Expression of MMP-9 in nasal mucosa tissue of patients in 3 groups detected by immunohistochemical staining (×400)

图2 Western blot 检测3组患者鼻黏膜/息肉组织中MMP-9和TGF-β1表达

Fig.2 Western blot analysis of MMP-9 and TGF-β1 expression in nasal mucosa/polyp tissue of patients in 3 groups

表1 3组患者MMP-9 mRNA及蛋白、TGF-β1蛋白表达水平变化 ($\bar{x} \pm s$)

Tab.1 Changes in expression levels of MMP-9 mRNA and protein and TGF-β1 protein in the 3 groups

组别	n	MMP-9 mRNA	MMP-9蛋白	TGF-β1蛋白
对照组	8	1.00±0.03	1.00±0.02	0.21±0.03
CRSsNP组	20	1.37±0.13^a	1.14±0.06^a	0.33±0.04^a
CRSwNP组	22	1.67±0.24^ab	1.98±0.37^ab	0.68±0.15^ab
F		42.967^＊＊	76.995^＊＊	87.763^＊＊

图3 细胞免疫荧光染色检测MMP-9表达（×400）

Fig.3 Detection of MMP-9 expression by cellular immunofluorescence staining (×400)

图4 Western blot 检测3组细胞中MMP-9、TGF-β1和EMT相关蛋白表达 A：Control组；B：TGF-β1组；C：TGF-β1+si-MMP-9组。

Fig.4 Western blot analysis of expression levels of MMP-9, TGF-β1 and EMT-related proteins in the 3 groups of cells

表2 3组细胞中MMP-9、TGF-β1和EMT相关蛋白表达水平比较（$\bar{x} \pm s$）

Tab.2 Comparison of expression levels of MMP-9, TGF-β1 and EMT-related proteins between 3 groups of cells

组别	n	TGF-β1	MMP-9	E-cadherin	vimentin	α-SMA
Control组	3	0.18±0.03	0.16±0.02	0.69±0.03	0.25±0.10	0.13±0.08
TGF-β1组	3	0.74±0.16^a	0.63±0.08^a	0.29±0.06^a	0.61±0.11^a	0.64±0.16^a
TGF-β1+si-MMP-9组	3	0.13±0.04^b	0.17±0.03^b	0.48±0.04^b	0.27±0.06^b	0.27±0.09^b
F		36.737^＊＊	84.273^＊＊	64.697^＊＊	14.435^＊＊	15.584^＊＊

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慢性鼻窦炎患者MMP-9的表达及与上皮间质转化的相关性研究

Expression of MMP-9 in patients with chronic rhinosinusitis and its correlation with epithelial-mesenchymal transition

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