血红素氧合酶1在鼻息肉组织中的表达及糖皮质激素的调节作用

摘要

目的 检测抗氧化应激反应的血红素氧合酶1(Heme oxygenase-1,HO-1)在慢性鼻-鼻窦炎伴鼻息肉(CRSwNP)患者鼻息肉组织中的表达及可能的调节机制.方法 收集25例CRSwNP患者的鼻息肉组织,并选取19例鼻中隔偏曲患者的钩突黏膜组织作为对照组.采取实时定量反转录聚合酶链反应(quantitative real-time PCR,qRT-PCR)、免疫组化及Western blot检测鼻息肉及钩突组织中HO-1的表达.另外选取15例鼻中隔偏曲患者的钩突黏膜组织进行组织培养,检测不同细胞因子和糖皮质激素对鼻黏膜组织中HO-1 mRNA的调节作用.以SPSS 20.0软件进行统计学分析.结果 HO-1 mRNA和蛋白质在鼻息肉组织中表达水平为1.220±0.397、1.409 ±0.701,明显高于对照组的0.464±0.318、0.017 ±0.114,差异有统计学意义(U值分别为22.00、1.00,P值均＜0.05).免疫组化结果显示HO-1在鼻息肉组织中的黏膜上皮细胞、黏膜下腺体细胞及炎症细胞上均有表达.体外组织刺激试验提示白细胞介素(IL) 17A刺激后HO-1 mRNA表达上调,刺激前为1.000,不同浓度IL-17A刺激后分别为17.264±4.275、19.128±4.605,差异有统计学意义(U=0,P＜0.05).地塞米松刺激后,HO-1 mRNA明显下调,刺激前为1.000,不同浓度地塞米松刺激后分别为0.370±0.101、0.316 ±0.167,差异有统计学意义(U=0,P＜0.05).转化生长因子β1(TGF-β1)刺激后,HO-1mRNA表达水平下调,刺激前后分别为0.217±0.322、0.070±0.070,差异有统计学意义(U=0,P＜0.05).结论 鼻息肉组织中HO-1表达上调,并受糖皮质激素抑制,提示HO-1可能可以作为糖皮质激素干预的一个靶点.%Objective To evaluate the expression and possible modulation of heme oxygenase-1 (HO-1) in nasal polyps of patients with chronic rhinosinusitis with nasal polyps (CRSwNP).Methods Nasal polyps and uncinate process tissues were collected from 25 CRSwNP patients and 19 healthy controls with nasal septal deviation.HO-1 expression was examined using qRT-PCR, immunohistochemistric staining and Western blot analysis.Moreover, additional uncinate process mucosal samples of 15 healthy controls with nasal septal deviation were harvested for nasal explant culture experiments.HO-1 expression was measured in cultured nasal explant in response to specific inflammatory and glucocorticoid stimulation.SPSS 20.0 software was used to analyze the data.Results The mRNA and protein expression of HO-1 was significantly increased in polyp tissues, 1.220 ± 0.397 in mRNA and 1.409 ± 0.701 in protein, compared with healthy controls 0.464 ± 0.318 in mRNA and 0.017 ± 0.1147 in protein (U =22.00 in mRNA and U =1.00 in protein, both P ＜ 0.05).The immunohistochemical results showed that HO-1 was mainly distributed in the epithelial layer, submucosal glands and inflammatory cells in nasal tissues.Nasal explant culture experiments demonstrated that HO-1 mRNA was upregulated by IL-17A.The HO-1 mRNA level before the stimulation was 1.000, and 17.264 ±4.275 after the stimulation of 1ng/ml IL-17A (U =0, P ＜ 0.05), 19.128 ±4.605 after the stimulation of l0 ng/ml IL-17A (U =0, P ＜0.05), but was significantly suppressed after stimulation with glucocorticoids (dexamethasone, DEX).The mRNA level after the glucocorticoids stimulation was 0.370 ± 0.101 (U =0, P ＜ 0.05) and 0.316 ± 0.167 (U =0, P ＜ 0.05)respectively.Furthermore, the HO-1 mRNA was inhibited by TGF-β1, the mRNA level was 0.217 ±0.322 (U =0, P ＜ 0.05), 0.070 ± 0.070 (U =0, P ＜ 0.05), respectively.Conclusion Increased HO-1 expression may play a role in the pathogenesis of CRSwNP, which may be considered as the therapeutic target.