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

摘要

Objective To explore ihe proleclion of siRNA LargeLing NF - kB P65 from sepsic acule liver injury in mice. Methods 70 male Kunming mice were randomly divided inlo heallhy conlrol group,sepsis group, specific interfering group and scrambled conlrol group, and ihe laller ihree groups were divided inlo posl - operalional 6 and 12 hours subgroups, each of which consisted of 10 mice. Relrovirus veclors which contained NF - kB P65 siRNA were adminislered Lo mice by caudal veins in ihe specific interfering group, relrovirus veclors containing scrambled siRNA lo ihe scrambled conlrol group, and normal saline of ihe same volume lo ihe heallhy conlrol group and ihe sepsis group. 1 hour laler, a mouse model of sepsic acule liver injury was buill by ihe meaning of cecal ligalion punclure ( CLP) in ihe Iwo virus groups and ihe sepsis conlrol group. Al posl - operalional hours 6 and 12, ihe experimental mice were sacrificed and liver lissue samples were collected. Hislopathologic changes were graded; level of NF - kB P65 prolein and expression of TNF - a. mRNA and protein were delected. Results The mouse model of sepsic acule liver injury was constructed successfully by the melhod of CLP. The expression level of NF - kB P65 and the liver injury of experimenlal mice were lightened by specific NF - kB P65 siRNA al post - operalional hours 6 and 12; the level of TNF - a mRNA and prolein declined ( P < 0. 05 ) at posl - operational 6 h in ihe specific interfering group. Conclusion The technology of small interfering RNA largeting NF - kB P65 could depress ihe expression of NF - kB , and further inhibil the excessive inflammalory reaction in sepsis, which lightened acute liver injury.%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.