降钙素基因相关肽转基因对同种异体移植血管病变的防治作用  

Prevention of allograft vasculopathy by transfection of calcitonin gene-related peptide

作　　者：张小宁[1] 陈道中[1] 郑宇辉[2] 梁建钢[2] 杨焕星[2] 林小文[2] 周小玲[3] 郑大利[4] 

机构地区：[1]福建医科大学附属协和医院心外科,福州350001 [2]福建医科大学附属协和医院病理科,福州350001 [3]福建医科大学附属协和医院中心实验室,福州350001 [4]福建医科大学基础医学院

出　　处：《中华医学杂志》2008年第45期3217-3221,共5页National Medical Journal of China

基　　金：福建省自然科学基金资助项目（2006j0094）

摘　　要：目的研究降钙素基因相关肽（CGRP）转基因对同种异体移植血管病变（AV）的防治作用，探讨其作用机制。方法大鼠分3组：（1）转染AdS—CGRP—EGFP组（A组），（2）转染AdS—EGFP组（B组），（3）未转染基因组（C组）。分别于术后4周、8周取标本，行病理、细胞凋亡检查，观察CGRP、EGFP基因及VCAM-1、iNOS表达。结果A组术后4周CGRP表达阳性，A组、B组术后4周EGFP基因表达阳性。A组术后4周、8周管腔闭塞指数低于B组和C组。A组术后d周细胞凋亡指数（AI）低于B组和C组。免疫组化显示，A组术后4周VCAM-1表达低于B组和C组，A组iNOS表达仅术后4周内膜低于B组和C组。结论CGRP基因转染后有效地防治了术后4周AV。Objective To explore suppression of allograft vasculopathy （AV） by transfer of the calcitonin gene-related peptide （CGRP）. Methods The descending thoracic aortas from 39 Lewis rats were grafted to the abdominal aortas of 39 F344 rats, and then the recipient rats were randomized into 3 groups ： Group A transfected with AdS-CGRP-EGFP, a gene construct containing sequences from the adenoviral oncoprotein, CGRP, and enhanced green fluorescent protein , Group B, transfected with AdS-EGFP containing sequences from the adenoviral oncoprotein and enhanced green fluorescent protein, and Group C without transfection. Four and 8 weeks later the abdominal aortas of 6 recipient rats from each were collected respectively. The degree of vascular obstruction was observed by microscopy. Frozen tissue sections were made and the inverted phase contrast fluorescence microscopy was used to observe the green fluorescence showing the expression of CGRP and enhanced green fluorescent protein （EGFP）. RT-PCR was used to detect the expression of CGRP. Immunohistochemistry was used to examine the expression of inducible nitric oxide synthase （iNOS） and vascular cell adhesion molecule-1 （VCAM-1）. The apoptosis index （AI） was detected by TUNEL method. Results CGRP expression was positive in both Group A and B 4 weeks later and negative in both groups 8 weeks later. Group A is approximately normal in pathological morphology 4 weeks later. The vascular luminal occlusion score of Group A was lower than Groups B and C 4 weeks later, and significantly higher than that Ⅰ the same group （ P 〈 0.05 ）, however, still lower than those of Groups B and C 8 weeks later. The AI 4 weeks later of Group A was significantly lower than those of Groups B and group C, however, there was no significance in AI among the 3 groups. Four weeks later the VCAM-1 expression levels in the tunicae intima, media, and adventita of Group A were all significantly lower than those of Group B C 4 weeks later, however, the iNOS expression level o

关 键 词：移植 同种 血管疾病 血管细胞黏附分子-1 一氧化氮合酶 

分 类 号：R686[医药卫生—骨科学]