CTGF表达在大鼠放射性肺纤维化中的作用  

Effect of connective tissue growth factor (CTGF) expression on radiation pulmonary fibrosis in rats

作　　者：黄山英[1] 宋良文[1] 张勇[1] 孙丽[1] 李杨[1] 

机构地区：[1]北京放射医学研究所病理研究室,100850

出　　处：《中华放射医学与防护杂志》2005年第3期213-216,共4页Chinese Journal of Radiological Medicine and Protection

基　　金：国家自然科学基金资助项目(30170292);军事医学科学研究基金资助项目(2002)

摘　　要：目的本研究结合体内外实验探讨CTGF在放射性肺纤维化(RPF)发生过程中的作用及与α-SMA之间的相互关系。方法应用MTT比色法检测60Coγ射线照射对人肺成纤维细胞(HLF)的促增殖作用,用Westernblot法观察CTGF和α-SMA在HLF中的表达,苦味酸天狼星红偏振光显微镜法检测Ⅰ、Ⅲ型胶原纤维的增减变化,免疫组织化学图像分析观察CTGF和α-SMA在RPF过程中的表达变化。结果体外实验显示,5Gy60Coγ射线照射能促进HLF增殖,并转化成肌成纤维细胞(MFb);CTGF蛋白合成增加,12h达高峰;α-SMA蛋白合成增加,48h处于高峰状态。体内实验显示,照射后1～3个月大鼠肺组织有少量胶原产生,以Ⅲ型胶原为主;6个月肺组织大量胶原产生,以Ⅰ型胶原为主;CTGF于照后1周在增生的大鼠肺Fb中开始合成,随照后时间延长而逐渐增强,3个月时最明显。照后1周可见增生的MFb中有α-SMA表达,随照后时间延长逐渐增多,3个月时达高峰。结论照射能诱导CTGF表达增高,后者能促进肺Fb向MFb转化,增强合成和分泌Ⅰ、Ⅲ型胶原的能力,参与了放射性肺纤维化的发生发展。Objective To explore the effect of connective tissue growth factor(CTGF) on initiation of radiation pulmonary fibrosis (RPF) and the relation to α_smooth muscle actin (α_SMA). Methods The promotive effect of CTGF on proliferation of human lung fibroblasts (HLF) by 5 Gy of ()^(60)Co γ_rays was determined by MTT colorimetry.The expressions of CTGF and α_SMA in HLF were observed by Western blot.Changes of collagen Ⅰ and Ⅲ in rat lungs were determined by Sirius red staining and polarization microscopy.Expressions of CTGF and α_SMA in RPF were observed with immuneohisto_chemistry and analysis. Results Expressions of CTGF and α_SMA were increased.CTGF reached its peak at 24 h after irradiation,whereas α_SMA still kept at a highlevel 72 h after irradiation.A small amount of collagen was produced in rat lung one month after irradiation,in which type Ⅲ collagen was the primary component.However,a large amount of collagen was produced in rat lung 3_6 months after irradiation,in which type Ⅰ was dominant.CTGF began to expression 1 week after irradiation in proliferative fibroblasts of rat lung,and it was most evident 3 months after irradiation.α_SMA began to express in proliferative myofibroblasts 1 week after irradiation, and the high peek was reached at 3 months after irradiation. Conclusion Irradiation can induce expression of CTGF in pulmonary tissue and the later can promote the transformation of fibroblasts to myofibroblasts,strengthen the ability of synthesis and secretion of type Ⅰ and Ⅲ collagen.;

关 键 词：放射性肺纤维化 CTGF ^60COΓ射线照射 苦味酸-天狼星红 人肺成纤维细胞 Western Α-SMA表达 MTT比色法 Ⅲ型胶原纤维 免疫组织化学 肌成纤维细胞 蛋白合成 时间延长 促增殖作用 BLOT法 大鼠肺组织 相互关系 发生过程 方法应用 

分 类 号：R818[医药卫生—放射医学]