心内直视术患儿体外循环前后静脉血白细胞基因表达的比较  

Effects of cardiopulmonary bypass on neutrophil gene expression in children

作　　者：王寿勇[1] 张诗海[1] 姚尚龙[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院麻醉科,武汉市430022

出　　处：《中华麻醉学杂志》2004年第8期565-569,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金资助项目(30271255)

摘　　要：目的 比较心内直视术患儿体外循环(CPB)前后静脉血白细胞基因表达的变化。方法择期CPB下行心内直视手术的患儿8例,分别在CPB前(主动脉置管时)和CPB停机时采取静脉血标本5ml,提取RNA。以RNA为模板逆转录获取cDNA;然后以cDNA为模板体外转录得到生物素标记的cRNA,cRNA经提纯和片断化后与u133A基因芯片杂交。杂交后的芯片用专用激光共聚焦扫描仪采集荧光强度值,并用Affymetrix(r)Microarray Suite 5.0软件分析基因表达规律。结果CPB前表达开放的基因共7023个,CPB后表达开放的基因7 810个。与CPB前比较,有2 873个基因出现差异性表达,1 390个增高,1 483个降低,差异性变化在3倍以上者共有96个,本研究按基因编码产物的功能对这部分基因进行了初步分析。结论 CPB引起心内直视术患儿静脉血白细胞基因出现明显的差异性表达,其特点与全身性炎性反应的临床认识相符,某些在功能上与肺损伤有关的基因表达异常升高。Objective Cardiopulmonary bypass (CPB) induces generalized inflammatory response. The aim of the study was to investigate the changes in neutrophil gene expression profile during CPB. Methods Eight pediatric patients with ventricular septal defect (5 males, 3 females) aged 3-8 yrs, weighing 14-20 kg undergoing open heart surgery under hypothermic (27-29 ℃) CPB were studied. Their cardiac function was well preserved (NYHA class Ⅰ- Ⅱ) . The surgery was performed under fentanyl anesthesia supplemented with low concentration of enflurane. Blood samples were taken before (at cannulation of aorta) and at the termination of CPB. A total RNA . isolation kit (RNeasy Maxi kit, QIAGEN) was used in isolating total RNA. cDNA was made through reverse transcription reaction under bootstrapping of poly-(dT) T7 primer, and then the cDNA was transcripted to cRNA. Biotin labeled UTP and CTP were used during the course of cRNA synthesis. After purification and fragmentation the biotin labeled cRNA was hybridized with U133A gene chips (Affymetrix). Results The number of the expression present genes (detected) significantly increased after CPB compared with that before CPB. A total of 2 873 genes showed differential expression after CPB, and 96 genes among them had their expression levels increase over 3 times after CPB. Conclusion CPB induces significant alterations in the expression of child neutrophil genes which are associated with a variety of functions including gene expression regulation, inflammatory and immune response, metabolism, hematopoisis, cell growth and apoptosis etc.

关 键 词：CPB 静脉血 血白细胞 心内直视术 体外循环 患儿 表达 RNA 基因 DNA 

分 类 号：R726.5[医药卫生—儿科]