体内转染核因子-κB诱捕物可减轻急性缺血性肾损伤  

Transfection of nuclear factor-κB decoy oligodeoxynucleotides prevents ischemic acute renal failure in rats

作　　者：曹长春[1] 丁小强[1] 欧周罗[2] 刘春凤[1] 李鹏[1] 王磊[1] 朱春芳[2] 

机构地区：[1]复旦大学附属中山医院肾内科,200032 [2]复旦大学上海医学院分子生物学与生物化学系

出　　处：《中华医学杂志》2003年第18期1597-1602,共6页National Medical Journal of China

基　　金：国家自然科学基金资助项目 (3 980 0 0 70 )

摘　　要：目的　探讨核因子 κB (NF κB)诱捕物寡聚核苷酸 (ODN)对急性缺血性肾损伤的保护作用。方法　采用肾动脉夹闭法制备大鼠缺血性急性肾衰竭 (iARF)模型 ,应用鱼精蛋白 脂质体法经肾动脉转染NF κB诱捕物ODN进行治疗。 2 4只大鼠被分成 4组 :假手术组 ,急性肾衰竭组 (iARF组 ) ,NF κB诱捕物ODN治疗组 (NF κB组 )和错配物ODN处理组。应用生化和组织学指标检测肾脏损伤的程度 ;凝胶电泳迟滞分析检测肾组织NF κB/DNA结合活性 ;免疫组化和RT PCR技术分别检测单核 巨噬细胞浸润 (M/MΦ)和单核细胞趋化蛋白 1(MCP 1)的表达。结果　经鱼精蛋白 脂质体法转染NF κB诱捕物ODN 12h后 ,ODN主要分布在肾小管上皮。与假手术组比较 ,iARF组血清Cr、BUN水平分别增加 10倍和 5倍 ( 2 5 6 μmol/L± 84 μmol/Lvs 2 5 μmol/L± 5 μmol/L和 4 3 4 7μmol/L± 13 4 8μmol/Lvs8 4 5mmol/L± 1 0 7mmol/L ,Ps<0 0 1) ;肾小管损伤评分明显升高 ( 3 6 3± 0 15vs 0 0 0± 0 0 0 ,P <0 0 1) ;NF κB /DNA结合活性明显增加 [中位数 (M) :1 75vs0 15 ,P <0 0 5 ];M/MΦ以及MCP 1的表达水平明显上升。与iARF组比较 ,NF κB组经诱捕物ODN治疗后 ,血清Cr水平下降 70 % ( 79μmol/L± 2 1μmol/Lvs 2 5 6 μmol/L± 84Objective To investigate the effect of nuclear factor κB (NF κB)decoy oligodeoxynucletides (ODN) on acute ischemic renal failure (iARF). Methods The right kidney was resected and the left renal artery was isolated so as to establish the animal model of iARF in 18 SD rats. Then the 18 rats were divided into 3 groups of 6 rats to be infused into the left renal artery with protamine liposome (PL) wrapped NF κB decoy ODN ((NF κB group), scrambled ODN (scrambled group), or nothing (iARF group) and then underwent clamping of the left renal artery for 60 minutes. Another six rats underwent sham operation with infusion of normal saline and without clamping of the renal artery (sham group). Twenty four hours later, blood was extracted from the inferior vena cava to detect the serum creatinine (Cr) and blood urine nitrogen (BUN). Normal saline was infused into the aorta to lavage the kidney. Then the kidney was taken to undergo histologic examination and examination of NF κB/DNA binding activity, monocyte/macrophage (M/MΦ) infiltration and MCP 1 gene and protein expression. Results In the rats injected with PL wrapped NF κB decoy ODN fluorescence was mainly distributed in the glomeruli 2 hours after injection of NF κB decoy ODN, mainly in the renal tubules 12 hours later, and remarkably subsided after 24 hours. In the rats injected with un wrapped NF κB decoy ODN, no fluorescence was seen at any time point. After 24 h of reperfusion, compared with sham operated animals, the serum Scr and BUN levels of the iARF group were about 10 times and 5 times those of the sham operation group (256 μmol/L±84 μmol/L vs. 25 μmol/L±5 μmol/L and 43 mmol/L±13 mmol/L vs. 8.45 mmol/L±1.07 mmol/L respectively. both P <0.001), the NF κB/DNA binding activity was markedly elevated [with the median values 1.75 vs.0.15 relative density unit (RDU), P <0.005], the renal tubular damage score was significantly higher (3.63±0.15 vs. 0.00±0.00 scores, P <0.01), M/MΦ infiltration and the expression of MCP 1

关 键 词：急性肾功能衰竭 NF-KB 单核细胞趋化蛋白-1 寡聚核苷酸 治疗 

分 类 号：R692.5[医药卫生—泌尿科学]