摘要
目的通过测定支气管哮喘(简称哮喘)大鼠肺组织中钙调神经磷酸酶(Calcineurin,CaN)蛋白、mRNA的表达及CaN的活性,探讨CaN在哮喘大鼠气道重塑中的作用。方法将20只Wistar大鼠随机分为哮喘组和对照组,每组10只。鸡卵清白蛋白溶液致敏及激发复制大鼠哮喘模型。HE染色观察气道炎症情况;图像分析观察支气管管壁厚度;实时定量(Real-time)PCR测定肺组织中CaNmRNA水平;免疫组织化学法检测肺组织中CaN蛋白的相对表达量;生物化学法检测大鼠肺组织CaN活性;流式细胞仪测定外周血单个核细胞细胞周期时相分布;ELISA法测定血浆中白介素4(IL-4)和肿瘤坏死因子α(TNF-α)的含量。结果①哮喘组大鼠支气管管壁厚度较对照组明显增加(P<0.01);②哮喘组大鼠肺组织中CaNmRNA水平的相对表达量、CaN蛋白的相对表达量及CaN的活性均较对照组高(分别P<0.05、P<0.01和P<0.01);③哮喘组大鼠血浆中IL-4和TNF-α含量较对照组明显增加(分别P<0.01,P<0.05);④与对照组相比,哮喘组大鼠外周血单个核细胞处于G0/G1期的百分率降低,处于S期、S+G2/M期百分率增加(P值均<0.01);⑤大鼠肺组织CaN活性分别与大鼠支气管管壁厚度呈正相关(P<0.01),与处于S期、S+G2/M期的外周血单个核细胞百分率呈正相关(P值均<0.05),与血浆中IL-4的含量呈正相关(P<0.05);血浆中IL-4含量与TNF-α含量及大鼠支气管管壁厚度亦呈正相关(P值均<0.05)。结论哮喘大鼠肺组织CaN的活性增加,其可能通过促进外周血单个核细胞的分裂、增殖及活化产生IL-4和TNF-α而参与哮喘气道重塑的形成。
Objective To investigate the expreession and activity of calcineurin（CaN） in asthmatic rat lungs,and to explore the role of CaN in airway remodeling with asthma. Methods Twenty Wistar rats were randomly divided into two groups：the control group and the asthma group, 10 rats each group. The rats were sensitized and challenged with ovalbumin to establish the asthmatic model, so did the rats in control group with saline instead. The following parameters were measured： airway inflammation by HE staining,the bronchial wall thickness by computer-assisted image analysis system; the expression of CaNmRNA by Real-time PCR,the protein expression of CaN by immunohistochemistry and the activity of CaN by biochemical method;the cell cycle distribution of monocyte in peripheral blood by flow cytomelry and the level of IL-4 and TNF-α by ELISA. Results ①The bronchial wall thickness of the asthma group was significantly increased compared with that of the control group （ P 〈0.01）. ②The expression of CaNmRNA,the protein expression of CaN and the activity of CaN in the asthma group were significantly higher than those of the control group（ P 〈0.05, P 〈0.01, P 〈0.01,respectively）. ③The level of IL-4 and TNF-α in plasma of asthma group was significantly higher than that in control group （ P 〈0.01, P 〈 0.05 ,respectively）. ④When compared with the control group, the percentage of monocyte in the G0/G1 phase was lower in the asthma group. Howover,the percentage of monocyte in the S phase and S＋G2/M phase was significantly higher （all P 〈0.01）. ⑤There was a positive correlation between the activity of CaN in rat lungs and the percentage of monocyte in the S phase and S＋G2/M phase in peripheral blood （all P 〈0.05） ,the same between the activity of CaN and the level of IL-4 in plasma or the bronchial wall thickness（ P 〈0.05, P 〈0.01, respectively）. There was a positive correlation between the level of IL-4 and the level of TNF-α in plasma（ P 〈0.05） ,and so between the level of IL -4 in plasma and the bronchial wall thickness （ P 〈0.05）. Conclusions The activity of CaN in asthmatic rat lungs was increased and the rising of it might play an important part in the development of airway remodeling in asthma by promoting the monocyte division and hyperplasia,or even causing the producing of IL-4 or TNF-α.
出处
《中华哮喘杂志（电子版）》
CAS
2010年第2期77-80,共4页
Chinese Journal of Asthma(Electronic Version)
