摘要
目的 评价磷脂酰肌醇-3激酶（PI3K）/蛋白激酶B（Akt）/核因子E2相关因子2（Nrf2）信号通路在兔内毒素休克诱发急性肺损伤中的作用.方法 健康雄性新西兰大白兔30只,2月龄,体重1.5～2.0 kg,采用随机数字表法分为3组（n=10）：对照组（C组）、急性肺损伤组（ALI组）和渥曼青霉素组（W组）.W组耳缘静脉注射渥曼青霉素0.6 mg/kg（溶于0.08 ml/kg二甲基亚砜中）,C组和ALI组耳缘静脉注射等容量生理盐水.30 min后,ALI组和W组耳缘静脉注射脂多糖5 mg/kg（溶于2 ml生理盐水中）制备急性肺损伤模型,C组耳缘静脉注射等容量生理盐水.注射脂多糖或生理盐水后6h时处死动物,取肺组织,光镜下观察病理学结果,并进行肺损伤评分,测定肺组织湿重/干重比值（W/D比值）,采用Western blot法测定肺组织磷酸化Akt（p-Akt）、Nrf2和血红素氧合酶-1（HO-1）的表达,采用实时荧光定量PCR法测定肺组织Nrf2 mRNA和HO-1 mRNA的表达.结果 与C组比较,ALI组和W组肺损伤评分和肺组织W/D比值升高,p-Akt、Nrf2及其mRNA和HO-1及其mRNA表达上调（P＜0.05）;与ALI组比较,W组肺损伤评分和肺组织W/D比值升高,p-Akt、Nrf2及其mRNA和HO-1及其mRNA表达上调（P＜0.05）.结论 PI3K/Akt/Nrf2信号通路激活是兔内毒素休克诱发急性肺损伤时机体的适应性调节机制.
Objective To evaluate the role of phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/nuclear factor E2-related factor 2 （Nrf2） signaling pathway in endotoxic shock-induced acute lung injury （ALI） in rabbits.Methods Thirty healthy male New Zealand white rabbits, aged 2 months, weighing 1.5-2.0 kg, were randomly divided into 3 groups （n =10 each） using a random number table： control group （group C）, ALI group, and wortmannin group （group W）.In group W, wortmannin 0.6 mg/kg （in 0.08 ml/kg dimethyl sulfoxide） was injected via the auricular vein, while the equal volume of normal saline was given in C and ALI groups.And 30 min later, lipopolysaccharide （LPS） 5 mg/kg （in 2 ml of normal saline） was injected via the auricular vein in ALI and W groups, while the equal volume of normal saline was given in group C.The rabbits were sacrificed at 6 h after LPS or normal saline injection.The lung was immediately removed for microscopic examination and for determination of wet/dry lung weight ratio （W/D ratio）, expression of phosphorylated Akt （p-Akt） , Nrf2 and heme oxygenase-1 （HO-1） （by Western blot）, and expression of Nrf2 and HO-1 mRNA （using fluorescent quantitative real-time reverse transcriptase-polymerase chain reaction）.The pathological changes of the lung were scored.Results Compared with group C, the lung injury scores and W/D ratio were significantly increased, and the expression of p-Akt, Nrf2 protein and mRNA, and HO-1 protein and mRNA was up-regulated in ALI and W groups （P〈0.05）.Compared with group ALI, the lung injury scores and W/D ratio were significantly increased,and the expression of p-Akt, Nrf2 protein and mRNA, and HO-1 protein and mRNA was up-regulated in group W （P〈0.05）.Conclusion Activation of PI3K/Akt/Nrf2 pathway is the regulatory mechanism of the body adapting to the development of endotoxic shock-induced ALI in rabbits.
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2015年第10期1257-1260,共4页
Chinese Journal of Anesthesiology
基金
国家自然科学基金（81372096）
天津市卫计委科技基金（2014KZ048）
