摘要
目的评价脓毒症小鼠急性肺损伤时血红素氧合酶-1(HO-1)表达与自噬的关系。
方法雄性C57BL/6小鼠48只,6~8周龄,体重20~25 g,采用随机数字表法分为4组(n=12):假手术组(SH组)、脓毒症组(CLP组)、HO-1激动剂Hemin+脓毒症组(Hemin+CLP组)和HO-1抑制剂SnPP+脓毒症组(SnPP+CLP组)。CLP组、Hemin+CLP组和SnPP+CLP组采用盲肠结扎穿孔法制备脓毒症模型。Hemin+CLP组及SnPP+CLP组于术前2 h分别腹腔注射Hemin 50 μmol/kg和SnPP 50 μmol/kg。术后24 h时颈动脉取血样,行血气分析,记录PaO2,计算氧合指数;取肺组织,光镜下行肺损伤评分,透射电镜下计数自噬小体,测定湿重/干重比值(W/D比值),采用Western blot法测定HO-1、Beclin-1和微管相关蛋白1轻链3Ⅱ型(LC3Ⅱ)的表达水平。
结果与SH组比较,CLP组、Hemin+CLP组及SnPP+CLP组氧合指数降低,肺组织W/D比值、自噬小体计数和肺损伤评分升高,CLP组和Hemin+CLP组肺组织HO-1、Beclin-1和LC3Ⅱ的表达上调(P〈0.05)。与CLP组比较,Hemin+CLP组氧合指数和肺组织自噬小体计数升高,W/D比值和肺损伤评分降低,肺组织HO-1、Beclin-1和LC3Ⅱ的表达上调,SnPP+CLP组氧合指数和肺组织自噬小体计数降低,W/D比值和肺损伤评分升高,HO-1、Beclin-1和LC3Ⅱ的表达下调(P〈0.05)。
结论HO-1表达上调介导自噬,参与了脓毒症小鼠急性肺损伤内源性保护机制。
Objective To evaluate the relationship between heme oxygenase-I (HO-1) expression and autophagy during acute lung injury in septic mice. Methods Forty-eight male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were randomly divided into 4 groups (n= 12 each) using a random number table: sham operation group (group SH) , sepsis group (group CLP) , HO-1 agonist hemin plus sepsis group (group Hemin+CLP) and HO-1 inhibitor SnPP plus sepsis group (group SnPP+CLP). Sepsis was induced by cecal ligation and puncture in CLP, Hemin+CLP and SnPP+CLP groups. Hemin 50 μmol/kg and SnPP 50 μmol/kg were intraperitoneally injected at 2 h before operation in Heroin+ CLP group and SnPP+CLP group, respectively. Arterial blood samples were taken at 24 h after operation for blood gas a- nalysis, PaO2 was recorded, and oxygenation index was calculated. Lungs were removed for examination of pathological changes which were scored and for determination of autophagosome count (with an electron mi- croscope) , weight/dry weight ratio (W/D ratio) and expression of HO-1, Becling-1 and microtubule-asso-ciated protein 1 light chain 3 11 (LC3 ]I ) in lung tissues (by Western blot). Results Compared with group SH, oxygenation index was significantly decreased, and W/D ratio, autophagosome count and patho- logical scores were increased in C LP, Hemin+CLP and SnPP+CLP groups, and the expression of HO-1, Beclin-1 and LC3 Ⅱ was significantly up-regulated in CLP and Hemin+CLP groups (P〈0. 05). Compared with group CLP, oxygenation index and autophagosome count were significantly increased, W/D ratio and pathological scores were decreased, and the expression of HO-1, Beelin-1 and LC3 Ⅱwas up-regulated in group Hemin+CLP, and oxygenation index and autophagosome count were significantly decreased, W/D ra- tio and pathological scores were increased, and the expression of HO-1, Beclin-1 and LC3 II was down- regulated in group SnPP+CLP (P〈O. 05). Conclusion Up-regulated expression of HO-1 mediates autoph- agy, which is involved in the endogenous protective mechanism underlvinz acute lunz iniurv in sentic mice.
出处
《中华麻醉学杂志》
CSCD
北大核心
2017年第9期1145-1148,共4页
Chinese Journal of Anesthesiology
基金
天津市滨海新区卫生计生委科技项目(2015BWKL001)