经肾动脉灌注细胞间粘附分子－1小片段干扰RNA减轻移植肾缺血再灌注损伤  

A novel deliver way of gene silencing by renal artery infusion of naked small interference RNA in a rat renal transplant model

作　　者：罗光恒[1] 卢一平[1] 杨立[1] 宋珺[1] 马学[1] 张立元[1] 李幼平[1] 

机构地区：[1]四川大学华西医院移植中心,成都610041

出　　处：《中华器官移植杂志》2009年第1期10-13,共4页Chinese Journal of Organ Transplantation

基　　金：国家973重大课题基金（2003CB515504）;教育部博士点基金（JYB00402051025）

摘　　要：目的探讨经肾动脉灌注细胞间粘附分子1小片段干扰RNA（ICAM-1 siRNA）对移植肾缺血再灌注损伤（IRI）的影响。方法将Fiskr大鼠随机分为3组．接受原位肾移植。供肾在冷保存1h后，按分组要求分别经肾动脉灌注生理盐水（NS组）、ICAM-1 siRNA（siRNA组）和完全错配siRNA（siRNA错配组），然后进行移植，切除受者自身双肾。分别于移植后6、12、24、48和72h处死大鼠，测定血清肌酐（Cr），观察移植肾组织学变化，测定移植肾组织中ICAM-1及其mRNA的表达情况。结果与NS组和siRNA错配组相比，siRNA组各时点肾小管坏死和炎症细胞浸润程度均较NS组和siRNA错配组有所减轻。siRNA组12、24、48h时的血Cr值明显低于NS组和siRNA错配组（P=0．01～0.001），尤以24h时的相差幅度最大。从术后12h起，siRNA组肾组织中ICAM-1 mRNA的表达明显低于NS组和siRNA错配组（P=0．01～0.001），24h时的差异达到最大，ICAM-1表达的变化与ICAM-1 mRNA一致。NS组与siRNA错配组相比较，各观察指标的差异均无统计学意义（P〉0．05）。结论移植前经肾动脉灌注ICAM-1 siRNA可减轻移植肾IRI，其机理可能与下调ICAM-1及其mRNA的表达有关。Objective To investigate the potential preventive role of small interfering RNA targeting ICAM-1 （ICAM-lsiRNA） infusion through renal artery in renal transplant cold ischemia injury model. Methods Fisher rats underwent renal isografting. Following removal, the left kidney was preserved in 0 to 4 ℃ heparin normal saline （NS） solution for 1 h to reinforce the cold ischemia injury. Before transplant, either ICAM lsiRNA （0.1 mg/kg）, or （NS） or control siRNA was infused through the renal artery with the renal vein clamped ex vivo. The left kidney was then implanted and the right kidney was removed. At 6, 12, 24, 48 and 72 h post operation, serum Cr was measured, and the severity of tubular necrosis and inflamed cells infiltration was assessed by histological grading scale respectively. The expression of ICAM-1 mRNA and protein in isograft was detected by real-time RT-PCR and Western blot, respectively. Results Compared with the NS and control siRNA groups, the values of serum Cr in siRNA group were lower at every time point, and the difference after 12 h was statistically significant （P = 0. 01-0. 001 ）, but there was no significant difference between NS and control siRNA groups （P〉0. 05）. The tubular necrosis and inflamed cells infiltration were less severe in siRNA group than those in NS and control siRNA groups, with the difference being significant especially in 12, 24 and 48 h. After 12 h, the expression levels of ICAM-1 mRNA and protein in the kidney of siRNA group were significantly down-regulated as compared with NS and control siRNA groups, and the difference reached its peak at 24 h. There was no significant difference among 3 groups at 72 h （P〉0.05）. Conclusion ICAM-1 siRNA can significantly prevent and attenuate the renal damage secondary to ischemia by down-regulating the expression of ICAM-1 protein and mRNA.

关 键 词：RNA干扰 胞间粘附分子1 再灌注损伤 肾 移植 

分 类 号：R686[医药卫生—骨科学]