慢性阻塞性肺部疾病大鼠支气管成纤维细胞体外培养条件下结缔组织生长因子水平检测及意义

目的:探讨慢性阻塞性肺部疾病(COPD)大鼠和正常大鼠支气管成纤维细胞体外培养下结缔组织生长因子(CTGF)的蛋白活性、转录水平以及胶原合成水平。方法:以正常大鼠(对照组)和COPD模型大鼠(实验组)为细胞来源,组织块法进行成支气管成纤维细胞原代培养,应用第5代细胞为研究对象。比色法进行培养液羟脯氨酸含量检测,免疫组织化学染色技术、RT-PCR、免疫印迹方法进行CTGF定位检测、蛋白活性及转录水平观察。结果:实验组羟脯氨酸含量显著高于对照组。对照组大鼠支气管成纤维细胞有一定水平的CTGF表达,实验组成纤维细胞CTGF阳性表达增强,免疫印记及RT-PCR结果显示CTGF蛋白活性增强、转录水平升高。结论:CTGF在成纤维细胞中的高表达是大鼠COPD发生发展的重要环节。

Expression and significance of connective tissue growth factor in fibroblast of bronchus of rats with chronic obstructive pulmonary diseases in vitro

Objective:To investigate the expression and clinical significance of connective tissue growth factor(CTGF)in fibroblast of bronchus of rats with chronic obstructive pulmonary disease(COPD)in vitro.Methods:Cultured fibroblasts were got from normal and COPD rats,respectively.The content of hydroxyproline was detected by colorimetry.Immunohistochemistry staining,Western Bolt and RT-PCR were used to examine the protein activity and mRNA level of CTGF.Results:The content of hydroxyproline in COPD group was higher than that in the control group.In the experiment group,CTGF expressed significantly,the protein activity and mRNA level of CTGF increased significantly with a time-dependence.Conclusion:The high level of expression of CTGF in the fibroblast maybe plays an important role in the occurrence and progress of COPD.