【摘要】 目的观察白细胞介素23(IL-23)基因沉默对肺组织IL-23蛋白表达、哮喘鼠气道炎症和支气管肺泡灌洗液(BALF)中干扰素(IFN-γ)、白细胞介素-5(IL-5)、肿瘤坏死因子-α(TNF-α)、细胞间黏附分子-1(ICAM-1)水平的影响.方法构建siRNA IL-23表达载体并通过脂质体转染获得高浓度的含有siRNA IL-23的浓缩液,RT-PCR和WB法检测IL-23mRNA及IL-23蛋白表达水平,确定转染成功.通过滴鼻方式吸入到卵蛋白(OVA)致敏的小鼠体内,观察其对BALF中炎症细胞的影响;ELISA方法检测支气管肺泡灌洗液(BALF)中细胞因子IFN-γ,IL-5,TNF-α,ICAM-1的变化;HE染色观察肺组织病理学改变;WB法检测肺组织中IL-23蛋白表达.结果与对照组相比,模型组及空质粒组BALF中IL-5,TNF-α,ICAM-1水平升高,IFN-γ水平下降,肺组织中IL-23蛋白表达量升高(P<0.01),沉默组上述各项指标明显下降,IFN-γ水平升高(P<0.01);病理学检测结果表明:与对照组相比,模型组和空质粒组气道内有大量炎性细胞浸润,沉默组炎症受到抑制(P<0.01).结论 IL-23在哮喘发病中起重要作用,阻断其基因表达可以明显改变哮喘症状.更多还原

【Abstract】 Objective To observe the effects of gene silencing of interleukin 23( IL-23) on IL-23 protein expression in lung tissue,airway inflammation in asthmatic rat and the levels of interferon-γ( IFN-γ),interleukin5( IL-5),tumor necrosis factor alpha( TNF-α),intercellular adhesion molecule-1( ICAM-1) in bronchoalveolar lavage fluid( BALF). Methods siRNA IL-23 expression vector was constructed and high concentration of siRNA containing IL-23 concentrate was gained by liposome transfection technology; RT-PCR and WB method were used to test the levels of IL-23 mRNA and IL-23 protein expression in order to identify the successful transfections.Mice was sensitized by intranasal ovalbumin( OVA) inhalation to observe its effects on inflammatory cells in BALF. Changes of IFN-γ,IL-5,TNF-α and ICAM-1 were detected via ELISA method. Pathological change was observed by HE staining and IL-23 protein expression was detected by WB in lung tissue. Results Compared with those in the control group,IL-5,TNF-α,ICAM-1 levels of the model group and the empty plasmid group increased,IFN-γ level declined in BALF and IL-23 protein expression in lung tissue rised( P < 0. 01). The above-mentioned indicators in silent group significantly decreased while IFN-γ level was enhanced( P < 0. 01).Pathological results showed that compared with the control group,model group and the empty plasmid group airway contained a large number of inflammatory cells infiltration,while the inflammation response in silent group was inhibited( P < 0. 01). Conclusion IL-23 may play an important role in the pathogenesis of asthma,blocking its gene expression could obviously relief the asthmatic symptoms.更多还原