降钙素基因相关肽体外转染对大鼠同种异体肺移植物缺血再灌注损伤的影响  

The effect of ex vivo adenovirus-mediated gene transfection of CGRP on the ischemical reperfusion injury of rat left lung allograft

作　　者：陈椿[1] 陈道中[3] 陈春美[4] 张小宁[2] 

机构地区：[1]福建医科大学附属协和医院胸外科, 福州350001 [2]广东省人民医院 [3]福建医科大学附属协和医院心外科, 福州350001 [4]福建医科大学附属协和医院神经外科, 福州350001

出　　处：《中华实验外科杂志》2008年第11期1469-1471,共3页Chinese Journal of Experimental Surgery

基　　金：福建省卫生厅百千万人才基金资助项目

摘　　要：目的观察和探讨腺病毒介导降钙素基因相关肽（CGRP）体外转染对大鼠同种异体肺移植物缺血再灌注损伤（IRI）的影响。方法SD大鼠建立同种异体左肺移植模型。设立假手术对照组（A组）、空白对照组（B组）、空载体对照组（C组）和基因转染组（D组）。D组供肺体外转染增强型绿色荧光蛋白标记降钙素基因相关肽重组腺病毒载体（A（15-CGRP-EGFP），颗粒滴度为3．2×10^9vp／ml。供肺保存6h行左肺原位移植。4h后检测动脉血气分析和气道内压（AWP）；荧光显微镜下观察移植肺组织绿色荧光变化；放射免疫法测定移植肺组织CGRP含量；检测移植肺湿-干重比率（W／D）比率、超氧化物歧化酶（SOD）活性、丙二醛（MDA）含量、髓过氧化物酶（MPO）活性；光镜观察移植肺病理组织学变化；酶联免疫吸附试验（ELISA）测定移植肺肿瘤坏死因子（TNF）-α、白细胞介素（IL）-10表达水平。结果B、C、D组出现明显IRI反应。但D组IRI反应明显减轻。荧光显微镜下C组和D组标本呈强绿色荧光反应。放射免疫法测定D组肺组织CGRP含量明显升高，AWP下降，PaO2上升、PaCO2下降、W／D比率下降，SOD活性增高，MDA含量和MPO活性下降；D组病理组织学检查示肺组织炎细胞浸润、水肿和渗出改变明显减轻；TNF-α表达水平下调、IL-10表达水平增强。结论CGRP体外转染可明显减轻IRI对供肺的损害，对大鼠肺移植的供肺起保护作用。Objective To investigate the effect of ex vivo adenovirus-mediated gene transfection of CGRP on ischemia-reperfusion injury （IRI） of rat left lung allograft. Methods A left lung isotransplantation formwork was constructed using SD rats. The SD rats were randomly divided into 4 groups ： A, the sham operation; B,the blank; C, blank adenovirus vector; D,the donor lung was retroperfused with Ads- CGRP-EGFP with the drop tite being 3.2×10^9 vp/ml. The donor lung which had been perfused was perserved for 6 h in the 4℃ LPDG fluid. Then the left lung orthotopic transplantation was performed. After 4 h, airway pressure was measured and arterial blood gas analysis was made to determine PaO2 and PaCO2. Lung allograft tissue was detected under the fluorescence microscopy to examine the green fluorescence changes. CGRP in lung tissue was assayed by radioactive immunoassay. The graft lung wet-to-dry （W/D） ratio, superoxide dismutase （SOD） activity, malondialdehyde （MDA） contents and myeloperoxidase （MPO） activity were detected. Graft pathohistological changes were examined under a light microscope. The expression of tumor necrosis factor （TNF）- α and interleukin （IL）-10 was quantified by ELISA. Resuits In groups B, C and D,all left lung allografts had IRI,but that in group D was obviously lightened. Specimens in group C and group D expressed strong green fluorescence under the fluorescence microscopy. The contents of CGRP in group D was obviously higher. The AWP,PaO2 and the activity of SOD were increased, but W/D ratio, the content of MDA and the activity of MPO decreased in group D. The lung tissue edema,interstitial inflammation and exudation in group D were obviously alleviated. Lower expression of TNF-α and higher expression of IL-10 were detected by ELISA. Conclusion Ex vivo adenovirus-mediated gene transfection of CGRP can obviously lighten IRI which can damage the donor lung, so can protect donor lung allograft from IRI.

关 键 词：降钙素基因相关肽 肺移植 再灌注损伤 基因转染 

分 类 号：R285.5[医药卫生—中药学]