靶向核转录因子-κB P65小干扰RNA对脓毒症所致小鼠急性肝损伤的影响  

Influence of siRNA targeting NF - κB P65 on sepsic acute liver injury in mice

作　　者：王君[1] 朱光发[1] 李丛锋[1] 靳丽妍[1] 吴春婷[1] 

机构地区：[1]首都医科大学附属北京安贞医院呼吸与重症医学科,北京100029

出　　处：《中国急救医学》2012年第2期108-111,I0002,共5页Chinese Journal of Critical Care Medicine

基　　金：2008年北京市教育委员会科技计划面上项目（No.KM200810025006）

摘　　要：目的 探讨靶向核转录因子（NF）-κB P65 小干扰RNA（siRNA）对脓毒症所致小鼠急性肝损伤的保护作用.方法 将70只雄性昆明小鼠随机分为四组：即健康对照组、脓毒症组、特异干扰组和乱序对照组,后三组每组均设置术后6、12 h两个时间点,每组每个时间点10只小鼠;术前1 h特异干扰组尾静脉注射NF-κB P65 siRNA逆转录病毒,乱序对照组注射Scrambled siRNA逆转录病毒,健康对照组及脓毒症组分别注射等体积生理盐水;除健康对照组小鼠均行盲肠结扎穿孔（CLP）法构建脓毒症急性肝损伤模型;于术后6、12 h留取肝组织标本,检测组织病理学改变,NF-κB P65蛋白表达水平,TNF-α mRNA及蛋白的表达水平.结果.与脓毒症组和乱序对照组比较,特异干扰组术后6、12 h肝内NF-κB P65蛋白的表达均降低,肝脏病理损害均减轻;特异干扰组术后6 h肝内TNF-α mRNA及蛋白水平显著降低（P〈0.05）.结论.靶向NF-κB P65 siRNA抑制NF-κB的表达后,能够抑制脓毒症所致过度炎症反应,减轻急性肝损伤.Objective To explore the protection of siRNA targeting NF - κB P65 from sepsic acute liver injury in mice. Methods 70 male Kunming mice were randomly divided into healthy control group ,sepsis group, specific interfering group and scrambled control group, and the latter three groups were divided into post - operational 6 and 12 hours subgroups, each of which consisted of 10 mice. Retrovirus vectors which contained NF-κB P65 siRNA were administered to mice by caudal veins in the specific interfering group, retrovirus vectors containing scrambled siRNA to the scrambled control group, and normal saline of the same volume to the healthy control group and the sepsis group. 1 hour later, a mouse model of sepsic acute liver injury was built by the meaning of cecal ligation puncture （CLP） in the two virus groups and the sepsis control group. At post - operational hours 6 and 12, the experimental mice were sacrificed and liver tissue samples were collected. Histopathologic changes were graded; level of NF - κB P65 protein and expression of TNF - a mRNA and protein were detected. Results The mouse model of sepsic acute liver injury was constructed successfully by the method of CLP. The expression level of NF - κB P65 and the liver injury of experimental mice were lightened by specific NF - κB P65 siRNA at post - operational hours 6 and 12 ; the level of TNF - a mRNA and protein declined （ P 〈 0.05 ） at post - operational 6 h in the specific interfering group. Conclusion The technology of small interfering RNA targeting NF - κB P65 could depress the expression of NF - κB, and further inhibit the excessive inflammatory reaction in sepsis, which lightened acute liver injury.

关 键 词：核转录因子-ΚB 小干扰RNA 逆转录病毒载体 急性肝损伤 

分 类 号：R692[医药卫生—泌尿科学]