ESTABLISHMENT OF NEW IMMUNOSUPPRESSIVE STRATEGY USING GENE TRANSFECTION TECHNIQUE

利用基因转染技术建立新的免疫抑制策略

基本信息

批准号：
12470274
负责人：
FUKUSHIMA Norihide
金额：
$ 8.7万
依托单位：
OSAKA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12470274/
关键词：
HEART TRANSPLANATION GENE TRANSFECTION END-STAGE CARDIAC FAILURE nFkB DECOY ORGAN PRESERVATION HVJ-LIPOSOME METHOD ISCHEMIA-REPERFSION INJURY 遺伝子導入法 HVJ-liposome法虚血耐性 nFkB-cis element decoy ラット拒絶耐性

项目摘要

BACKGROUND : Nuclear factor-kappaB (NFkappaB) is critical for the transcription of multiple genes involved in myocardial ischernia-reperfusion injury. Therefore, we hypothesized that blocking NFkappaB would attenuate ischemia-reperfusion injury after prolonged myocardial preservation, resulting in an improvement in cardiac function.METHODS : Double-stranded oligodeoxynucleotides with a specific affinity for NFkappaB (NFkappaB decoy group) or a "rambled decoy group were transfected into rat hearts using a hemagglutinating virus ofjapan-liposome method. After 16 "urs of preservation in Euro-Collins solution at 4 degrees C, the cardiac grafts were heterotopically transplanted into recipient rats of the same strain. RESULTS: Fluorescein isothiocyanatestaining showed introduction of double-stranded oligonucleotides into the nuclei of endothelial cells and cardiomyocytes. After 1 hour of reperfusion the NFkappaB decoy group showed significantly higher degrees of recovery of left ventricular function as well as significantly lower levels of serum creatine phosphokinase, myocardial water content, tissue 1L-8, and neutrophil infiltration than did the scrambled decoy group (p < 0.05). CONCLUSIONS : Gene transfection of the NFkappaB decoy attenuates ischemia-reperfusion injury after prolonged heart preservation. As a result, this method appears to be a novel strategy for enhanced myocardial preservation.

背景：核因子-kappaB (NFkappaB) 对于参与心肌缺血再灌注损伤的多个基因的转录至关重要。因此，我们假设阻断 NFkappaB 可以减轻长期心肌保存后的缺血再灌注损伤，从而改善心功能。方法：将具有 NFkappaB 特异性亲和力的双链寡脱氧核苷酸（NFkappaB 诱饵组）或“散乱诱饵组” 16“urs后使用日本血凝病毒-脂质体方法转染至大鼠心脏。 4℃保存于欧洲柯林斯溶液中后，将心脏移植物异位移植到同一品系的受体大鼠体内。结果：异硫氰酸荧光素染色显示双链寡核苷酸引入内皮细胞和心肌细胞的细胞核。再灌注1小时后，NFkappaB诱饵组的左心室功能恢复程度显着高于乱序诱饵组，并且血清肌酸磷酸激酶、心肌含水量、组织1L-8和中性粒细胞浸润水平显着低于乱序诱饵组（p < 0.05）。结论：NFkappaB 诱饵基因转染可减轻长期心脏保存后的缺血再灌注损伤。因此，该方法似乎是增强心肌保存的新策略。