分泌性中耳炎病灶组织中伴有Kazal基序富含半胱氨酸的逆转诱导蛋白的表达及其与细胞外基质重塑、骨破坏及细胞凋亡的相关性

Expression of reversion-inducing cysteine-rich protein with Kazal motifs in secretory otitis media lesion tissues and its correlation with extracellular matrix remodeling,bone destruction and cell apoptosis

目的探讨分泌性中耳炎病灶组织中伴有Kazal基序富含半胱氨酸的逆转诱导蛋白(RECK)的表达及其与细胞外基质重塑、骨破坏及细胞凋亡的相关性。方法选择58例接受开放式乳突改良根治手术或完壁式乳突根治手术的分泌性中耳炎患者,术后留取分泌性中耳炎病灶组织;另取同期因外伤接受外耳道成形术的44例患者,术后留取正常外耳道上皮组织。检测两种组织中以下指标的mRNA表达量:RECK、细胞外基质重塑分子[基质金属蛋白酶2(MMP2)、转化生长因子β1(TGF-β1)、Smad 2/3、基质金属蛋白酶组织抑制因子(TIMP)1、TIMP2]、骨破坏分子[Runt相关转录因子2(RUNX2)、骨保护素(OPG)、核因子κB受体活化因子(RANK)、RANK配体(RANKL)]及细胞凋亡基因[第10号染色体缺失的磷酸酶及张力蛋白同源的基因(PTEN)、含半胱氨酸的天冬氨酸蛋白水解酶3(Caspase-3)、Livin、细胞周期蛋白D1(Cyclin D1)、增殖细胞核抗原(PCNA)]。分析分泌性中耳炎局部组织中RECK mRNA表达量与细胞外基质重塑分子、骨破坏分子、细胞凋亡基因mRNA表达量的相关性。结果分泌性中耳炎病灶组织中RECK、TIMP1、TIMP2、RUNX2、OPG、Livin、Cyclin D1、PCNA的mRNA相对表达量均低于正常外耳道上皮组织,而MMP2、TGF-β1、Smad 2/3、RANK、RANKL、PTEN、Caspase-3的mRNA相对表达量均高于正常外耳道上皮组织(均P<0.05)。分泌性中耳炎病灶组织中RECK的相对mRNA表达量与TIMP1、TIMP2、RUNX2、OPG、Livin、Cyclin D1、PCNA的mRNA相对表达量呈正相关,与MMP2、TGF-β1、Smad 2/3、RANK、RANKL、PTEN、Caspase-3的mRNA相对表达量呈负相关(均P<0.05)。结论分泌性中耳炎病灶组织中RECK呈低表达,这能够促进细胞外基质重塑、骨质破坏及细胞凋亡的发生。

Objective To explore the expression of reversion-inducing cysteine-rich protein with Kazal motifs(RECK) in secretory otitis media lesion tissues and its correlation with extracellular matrix remodeling,bone destruction and cell apoptosis.Methods Fifty-eight secretory otitis media patients received modified open radical mastoidectomy or complete mastoidectomy were selected,and the secretory otitis media lesion tissues were retained after operation;forty-four patients undergoing reconstruction of the external auditory canal due to trauma were selected during the same period,and the epithelial tissues of normal external auditory canal were retained after operation.The mRNA expressions of following indicators in both types of tissues were detected,including RECK,extracellular matrix remodeling factors[matrix metalloproteinase 2(MMP2),transforming growth factor β1(TGF-β1),Smad 2/3,tissue inhibitor metalloproteinase(TIMP) 1,TIMP2],bone destruction factors[Runt related transcription factor 2(RUNX2),osteoprotegerin(OPG),receptor activator of nuclear factor-κ B(RANK),RANK ligand(RANKL)],and cell apoptosis gene[gene of phosphatase and tensin homolog deleted on chromosome 10(PTEN),cysteine aspartic acic specific protease-3(Caspase-3),Livin,Cyclin D1,proliferating cell nuclear antigen(PCNA)].The correlation of RECK mRNA expression in secretory otitis media lesion tissues with mRNA expressions of extracellular matrix remodeling factors,bone destruction factors and cell apoptosis gene was analyzed.Results Compared to the epithelial tissues of normal external auditory canal,the relative mRNA expressions of RECK,TIMP1,TIMP2,RUNX2,OPG,Livin,Cyclin D1 and PCNA decreased in secretory otitis media lesion tissues,whereas the relative mRNA expressions of MMP2,TGF-β1,Smad 2/3,RANK,RANKL,PTEN and Caspase-3 increased(all P<0.05).The relative mRNA expression of RECK in secretory otitis media lesion tissues was positively correlated with the relative mRNA expressions of TIMP1,TIMP2,RUNX2,OPG,Livin,Cyclin D1 and PCNA,while was negatively correlated with the relative mRNA expressions of MMP2,TGF-β1,Smad 2/3,RANK,RANKL,PTEN and Caspase-3(all P<0.05).Conclusion RECK in secretory otitis media lesion tissues exhibits a low-expression,which can promote the occurrence of extracellular matrix remodeling,bone destruction and cell apoptosis.