谷氨酰胺对脓毒症小鼠腹腔巨噬细胞细胞因子分泌的影响  

Effects of glutamine on inflammatory cytokine release in murine peritoneal macrophages during sepsis

作　　者：袁媛[1] 王学敏[1] 梁梦凡[1] 江伟[1] 

机构地区：[1]上海交通大学医学院第六人民医院麻醉科,上海200233

出　　处：《上海交通大学学报（医学版）》2008年第7期822-826,共5页Journal of Shanghai Jiao tong University:Medical Science

基　　金：上海市科委科研计划项目(054119516)~~

摘　　要：目的研究谷氨酰胺(Gln)体内作用对脓毒症小鼠腹腔巨噬细胞细胞因子分泌的影响。方法昆明小鼠60只,随机分为假手术组(Sham组,n=20)、手术对照组(Con组,n=20)、术后Gln处理组(Gln组,n=20)。Gln组和Con组小鼠采用盲肠结扎穿孔法制备脓毒症模型,建模后即刻经尾静脉分别注射Gln 0.75 g/kg和等量生理盐水;Sham组小鼠仅开腹翻动盲肠,术后处理同Con组。三组均于6 h后采血收集血清,腹腔灌洗分离巨噬细胞;ELISA法测定巨噬细胞裂解液及血清中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)和IL-10的水平;RT-PCR测定巨噬细胞TNF-αmRNA表达;Western blotting检测巨噬细胞热休克蛋白72(HSP72)表达。结果Gln组腹腔巨噬细胞裂解液中TNF-α和IL-6水平明显低于Con组(P<0.01或0.05),而与Sham组相比无显著差异(P>0.05)。Gln组血清中TNF-α水平明显低于Con组(P<0.05),但两组TNF-α、IL-6和IL-10水平均高于Sham组。Gln组小鼠腹腔巨噬细胞HSP 72表达较Con组和Sham组显著升高(P<0.01)。Con与Gln组巨噬细胞TNF-αmRNA表达无显著差异,但均明显高于Sham组(P<0.01)。结论Gln体内作用可以抑制脓毒症小鼠腹腔巨噬细胞释放TNF-α和IL-6,同时增加其HSP 72的表达。Objective To investigate the in vivo effects of glutamine （Gin） on inflammatory cytokine release in murine peritoneal macrophages during sepsis. Methods Sixty Kunming mice were randomly divided into sham-operation group （ Sham group, n = 20） , operation control group （ Con group, n = 20） and Gin-treatment group （ Gin group, n = 20 ）. Sepsis was induced by cecal ligation and puncture in Gin group and Con group, and Gin （0.75 g/kg） or saline was immediately administered via single tail vein injection. Serum was collected and macrophages were harvested from peritoneal lavage at 6 h in these three groups. Intracellular and serum cytokines of tumor necrosis factor-α（TNF-α） , interleukin（IL）-6 and IL-10 were detected by ELISA. The expression of TNF-α mRNA in macrophages was analyzed by RT-PCR, and the expression of heat shock protein （HSP） 72 in macrophages was evaluated by Western blotting. Results Gin group demonstrated significantly lower intracellular TNF-α and IL-6 levels than Con group（P 〈0.01 or 0.05） , while there was no significant difference between Gin group and Sham group （P 〉 0.05）. The serum TNF-α level was significantly lower in Gin group than in Con group（P 〈0.05）, while the levels of serum TNF-α, IL-6 and IL-10 in these two groups were higher than those in Sham group. Gin treatment led to significant HSP 72 expression in macrophages compared to Con group and Sham group（P 〈0.01 ）. There was no significant difference in the expression of TNF-α mRNA in macrophages between Con group and Gin group, while both were significantly higher than Sham group （P 〈0.0l ）. Conclusion Gin treatment significantly inhibits TNF-α and IL-6 release, and increases HSP 72 expression in murine peritoneal macrophages during sepsis.

关 键 词：谷氨酰胺 巨噬细胞 细胞因子 热休克蛋白 

分 类 号：R459.7[医药卫生—急诊医学]