目的探讨硫氧还蛋白结合蛋白/维生素D3上调蛋白1(VDUP1)在不同时期哮喘病人嗜酸细胞中的表达及其与哮喘临床症状的关系.方法抽取正常自愿受试者14例,急性发病未经任何治疗的16例轻到中度哮喘病人和通过吸入糖皮质激素而处于哮喘缓解期的35例病人的外周静脉血15 ml进行外周血嗜酸细胞计数和诱导痰嗜酸细胞计数,随后行肺功能检查测定FEV1.0%和PEF%.分离纯化静脉血嗜酸细胞,提取总RNA.RT-PCR同时扩增特异性片段VDUP1和内参β-Actin.取10μl反应物进行1%琼脂糖凝胶电泳,用Gel-Pro软件分析凝胶成像,测定VDUP1和β-Actin灰度值,求VDUP1/β-Actin比值,并比较不同组别之间VDUP1表达的变化,分析VDUP1/β-Actin比值与诱导痰嗜酸细胞、FEV1.0%、PEF%的相关性.结果急性期未经治疗的哮喘病人嗜酸细胞(0.314±0.242)与正常人比较(0.532±0.279)显著降低,而经吸入糖皮质激素处于缓解期哮喘病人嗜酸细胞VDUP1/β-Actin比值(0.612±0.381)与正常对照组无显著差异.在急性发作未经治疗哮喘组嗜酸细胞VDUP1表达与FEV1.0%(r=0.587,P=0.046)和PEF%(r=0.563,P=0.033)呈正相关,而与诱导痰嗜酸细胞计数呈负相关关系(r=-0.436,p=0.049).结论嗜酸细胞VDUP1的表达与哮喘嗜酸细胞活化相关,并影响哮喘病人的临床症状.%Objective To investigate the expression ofvitamine D3 upregulated protein-1 (VDUP1) in peripheral eosinophils at different stagesof asthma and its correlation with clinical manifestations of asthma. Methods Fourteen normal volunteers and 51 mild to moderate asthma patients, including 16 untreated patients with asthma attack and 35 with asthma remission by continuously corticosterone inhalation. The symptom severity and pulmonary function indices were evaluated and induced sputum eosinophil counts and blood eosinophil count measured. VDUP1 and β-actin gene fragments were amplified simultaneously by RT-PCR from the total RNA of peripheral eosinophils, and 10 μl of the RT-PCR product underwent agarose gel electrophoresis and the VDUP1/β-actin ratio was obtained by Gel-Pro software. Results VDUP1/β-actin ratio significantly decreased in untreated patients with asthma attacks in comparison with normal volunteers (0.314±0.242 vs 0.532±0.279) but not in patients with asthma remission (0.612 ±0.381). In the former patients, a positive correlation of VDUP1/β-actin ratio was found with FEV1.0% (r=0.587, P=0.046) and %PEF (r=0.563, P=0.033), whereas an inverse one observed with sputum eosinophil count (r=-0.436, P=0.049). Conclusion VDUP1 expression in the eosinophils correlates to eosinophil activation and may influence the disease severity of asthma patients.
展开▼
