靶向OX40基因的siRNA抑制大鼠肝脏移植排斥反应的研究  

Prolonged rat liver allograft survival by in vivo targeting OX40-siRNA OX40-OX40L co-stimulatory cascade blockade

作　　者：武聚山[1] 夏仁品[1] 卢实春[1] 张毅[1] 娄金丽[2] 李宁[1] 

机构地区：[1]首都医科大学附属北京佑安医院肝胆外科,100069 [2]北京市卫生局肝炎研究所

出　　处：《中华普通外科杂志》2008年第7期516-519,共4页Chinese Journal of General Surgery

基　　金：教育部留学回国人员科研启动基金（2007-1108）,首都医科大学基础与临床合作基金资助项目（2006JL55）

摘　　要：目的探讨RNA干扰阻断OX40-OX40L共刺激通路对大鼠移植肝存活时间的影响。方法制作原位肝脏移植物大鼠模型，供体DA大鼠，受体Lewis大鼠。移植术毕时将5ml含5×10^9pfu的pLVTHM-OX40-siRNA重组慢病毒经阴茎背静脉在10秒内注射受鼠体内，术后观察受鼠存活时间，移植肝脏进行组织病理学检查，采用ELISA法检测大鼠外周血IL-2、IFN-γ的水平，并进行受者大鼠脾细胞对供者大鼠脾细胞的混和淋巴细胞反应（MLR）。结果转染组大鼠肝脏移植物的存活时间为（74．0±9．3）d，明显长于对照组（7．3±0．5）d和未转染组（7．5±0．5）d；转染组移植肝组织炎细胞浸润、间质水肿、肝组织坏死程度较对照组和未转染组减轻；耐受组大鼠脾脏中T淋巴细胞刺激T淋巴细胞增殖的能力较对照组降低（t=25．1，P〈0．01）；移植术后第7d，转染组血清IL-2浓度为（46±8．4）pg／ml，IFN-γ浓度为（202．7±14．6）pg／ml，均显著低于对照组和未转染组（分别t=176．4，45．5，P〈0．01）。结论转染靶向OX40的siRNA，可通过阻断OX40-OX40L共刺激通路，抑制大鼠肝脏移植后的排斥反应，延长大鼠移植肝的存活时间。Objective To investigate the effect of blockading OX40-OX40L co-stimulatory signaling on the survival time of liver allograft in rat. Methods siRNA-expression vectors were constructed to targeting OX40. 3 -5 minutes before DA to Lewis orthotopic liver transplantation was performed, 5 × 10^9 pfu of targeting OX40 siRNA plasmid DNA were diluted in 5 ml of phosphate buffered saline （PBS） and injected intravenously into recipient Lewis rat over a period of 10 seconds. Serum IL-2 and IFN-γ levels were assayed by ELISA, and mix lymphocyte response （MLR） were tested by ^3H-thymidine. Results The survival time of recipients in siRNA treatment group （74. 0 ±9. 3 ） was significantly longer than that in control group [ （7.3 ± 0. 5 ） days ]. In experiment group, the inflammatory cell infiltration and liver tissue structure destruction were very slight. The concentration of serum IL-2 was much lower in siRNA treatment group [ （46 ± 8.4） pg/ml ] than that in control group [ （ 286. 5 ± 14. 6） pg/ml ]. Meanwhile, the concentration of serum IFN-γ was much lower in siRNA treatment group [ （202. 7 ± 14. 6） pg/ml] than that in control group [ （ 1682. 7 ± 87. 9 ） pg/ml ]. Conclusion Administration of OX40-siRNA can blockade OX40-OX40L co-stimulatory signaling pathway, hence inhibit the rejection of liver allograft.

关 键 词：肝移植 移植物排斥 OX40 小干扰RNA 

分 类 号：R686[医药卫生—骨科学]