碱性成纤维细胞生长因子反义寡核苷酸对大鼠肺纤维化的影响  

Effect of antisense oligodeoxynucleotide of basic fibroblast growth factor on rat model of pulmonary fibrosis

作　　者：谭维军[1] 程真顺[1] 谭秋月[1] 叶燕青[1] 江平[2] 

机构地区：[1]武汉大学中南医院呼吸内科,430071 [2]武汉大学中南医院普外科,430071

出　　处：《中华实验外科杂志》2014年第11期2490-2493,共4页Chinese Journal of Experimental Surgery

基　　金：湖北省自然科学基金资助项目（2012FFB04310）

摘　　要：目的 观察碱性成纤维细胞生长因子（bFGF）反义寡核苷酸对大鼠肺纤维化的影响.方法 将30只大鼠随机分为正常对照组、肺纤维化组、空载体转染组、反义bFGF转染组、正义bFGF转染组,每组6只.造模处理后,计算各组大鼠肺系数,比较肺组织纤维化程度,酶联免疫吸附试验（ELISA）检测血清和肺泡灌洗液bFGF含量及肺匀浆羟脯氨酸含量,逆转录-聚合酶链反应（RT-PCR）检测肺匀浆中bFGF、α-平滑肌肌动蛋白（α-SMA）、Ⅰ型胶原（Collagen Ⅰ）、转化生长因子-β1（TGF-β1）、结缔组织生长因子（CTGF）、Smad3和Smad7的mRNA表达.结果 对照组、肺纤维化组、空载体转染组、反义bFGF组、正义bFGF组大鼠肺系数值分别为：（7.5±1.5）％、（1O.8±1.4）％、（11.0±1.4）％、（8.8±1.7）％、（12.5±1.6）％;肺间质纤维化分级为：0、2.38±0.52、2.41±0.48、1.62±0.45、2.82±0.54;反义bFGF转染组肺系数明显降低（P＜0.05）,正义bFGF转染组肺系数明显增高（P＜0.05）;反义bFGF转染组肺间质纤维化明显减轻（P＜0.05）,正义bFGF转染组明显加重（P＜0.05）;反义bFGF转染组血清、肺泡灌洗液中bFGF含量及肺匀浆中羟脯氨酸含量明显降低（P＜0.05）,正义bFGF转染组含量明显增高（P＜0.05）;反义bFGF转染组肺匀浆中α-SMA、Collagen Ⅰ、TGF-β1、CTGF及Smad3 mRNA的表达水平明显降低（P＜0.05）,正义bFGF转染组表达明显增高（P＜0.05）,Smad7 mRNA的表达刚好相反.结论 bFGF反义寡核苷酸可以减轻博莱霉素诱导的大鼠肺纤维化,可能机制是bFGF反义寡核苷酸下调了TGF-β1 Smad信号通路.Objective To observe the effect of antisense oligodeoxynucleotide （AODN） of basic fibroblast growth factor （bFGF） on the rat models of pulmonary fibrosis.Methods Thirty rats were divided into five groups （6 rats of each group）：control group,pulmonary fibrosis group,empty vector transfection group,AODN group and RODN group.Lung coefficient was counted,pathologic changes in the lung tissue were observated to determine degree of pulmonary fibrosis,enzyme linked immunosorbent assay （ELISA） was used to determine the concentrations of hydroxyproline in lung tissue homogenate and the expression of bFGF in the serum and bronchoalveolar lavage fluid,and reverse transcription-polymerase chain reaction （RT-PCR） was employed to detect the mRNA expression of bFGF,α-smooth muscle actin （α-SMA）,type Ⅰ collagen,transforming growth factor-β1 （TGF-β1）,connective tissue growth factor （CTGF）,Smad3 and Smad7 in lung tissue homogenate.Results In control group,pulmonary fibrosis gToup,empty vector transfection group,AODN group and RODN group,the lung coefficient was 7.5 ± 1.5,10.8 ± 1.4,11.0 ± 1.4,8.8 ± 1.7 and 12.5 ± 1.6 respectively,the degree of pulmonary fibrosis was 0,2.38 ± 0.52,2.41 ± 0.48,1.62 ± 0.45 and 2.82 ±0.54 respectively,the synthesis of bFGF was significantly decreased in AODN group and significantly increased in RODN group （P 〈 0.05）,the concentration of HYP was markedly decreased in AODN group and markedly increased in RODN group （P 〈 0.05）,the mRNA expression of bFGF,α-SMA,type Ⅰ collagen,TGF-β1,CTGF and Smad3 was markedly decreased in AODN group and markedly increased RODN group （P 〈 0.05）,and that of Smad7 was markedly increased in AODN group and markedly decreased in RODN group.Conclusion：These results demonstrate that AODN of bFGF can inhibit the development of pulmonary fibrosis induced by Bleomycyn in rats possibly by down-regulating the TGF-β1-Smad signaling pathway.

关 键 词：碱性成纤维细胞生长因子 反义寡核苷酸 肺间质纤维化 转化生长因子-β1-Smad信号通路 

分 类 号：R363[医药卫生—病理学]