摘要
目的:探讨针对核因子-κB(NF-κB)P65的小干扰RNA(siRNA)在脓毒症急性肺损伤防治中的保护作用。方法:将体质量18 g～20 g,雄性,昆明小鼠70只,随机分为健康组、脓毒症组、特异干扰组及乱序对照组,后3组均设置术后6 h、12 h 2个亚组(每组10只)。术前1 h特异干扰组尾静脉注射NF-κB P65 siRNA反转录病毒,乱序对照组注射scrambled siRNA反转录病毒,健康组及脓毒症组注射等量0.9%氯化钠液;分别对脓毒症组、特异干扰组及乱序对照组小鼠行盲肠结扎穿孔(CLP)手术构建急性肺损伤模型;术后6 h、12 h留取肺组织标本,观察肺病理改变;湿干质量比值(W/D);肺NF-κB染色强度;NF-κB mRNA、基质金属蛋白酶-9 mRNA(MMP-9 mRNA)及蛋白的活性水平。结果:采用CLP法成功构建小鼠急性肺损伤模型。与脓毒症组和乱序对照组比较,特异干扰组术后6 h、12 h肺组织较同时段脓毒症及乱序对照组肺组织病理损害及W/D值,NF-κB P65 mRNA,MMP-9 mRNA及蛋白活性减低,其中在6 h差异有统计学意义(P<0.05)。结论:针对NF-κB P65的siRNA抑制NF-κB的表达后,能够抑制脓毒症所致的早期的过度炎症反应,减轻急性肺损伤。
Objective：To explore the protection of siRNA against nuclear factor-κB（NF-κB）P65 from septic acute lung 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 containing 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 septic acute lung 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 lung tissue samples were collected.Histopathologic changes,level of wet/dry ratio,NF-κB intensity by immunohistochemical staining,expression of NF-κB P65 mRNA,matrix metalloproteinase-9 mRNA（MMP-9 mRNA）,protein activity were detected.Results： The mouse model of septic acute lung injury was constructed successfully by the method of CLP.Compare to sepsis group and scrambled control group at the corresponding time,the expression level of NF-κB P65 mRNA,the lung injury of experimental mice,wet/dry ratio,the level of MMP-9 mRNA and protein activity declined,significant differences were seen at post-operational 6 h（P〈0.05）.Conclusion： The technology of small interfering RNA against NF-κB P65 could depress the expression of NF-κB,and further inhibit the early phasic excessive inflammatory reaction in sepsis,which lightened acute lung injury
出处
《心肺血管病杂志》
CAS
2012年第3期333-337,共5页
Journal of Cardiovascular and Pulmonary Diseases
基金
北京市教育委员会科技计划面上项目(编号:KM200810025006)
北京市自然科学基金项目(7112039)
