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.
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.