›› 2012, Vol. 32 ›› Issue (11): 1472-.doi: 10.3969/j.issn.1674-8115.2012.11.015

• 论著(基础研究) • 上一篇    下一篇

谷氨酰胺对脓毒血症大鼠肠淋巴液抑制RAW264.7细胞分泌功能作用的影响

梁梦凡, 王学敏, 薛 瑛, 江 伟   

  1. 上海交通大学附属第六人民医院麻醉科/ICU, 上海 200233
  • 出版日期:2012-11-28 发布日期:2012-11-30
  • 通讯作者: 江 伟, 电子信箱: jiangw@sjtu.edu.cn。
  • 作者简介:梁梦凡(1981—), 女, 博士;电子信箱: celle707@163.com。
  • 基金资助:

    上海市科委科研计划项目(054119516)

Effects of glutamine on secretion suppression of RAW 264.7 cells induced by mesenteric lymph in rats with sepsis

LIANG Meng-fan, WANG Xue-min, XUE Ying, JIANG Wei   

  1. Department of Anesthesiology/ICU, the Sixth People's Hospital, Shanghai Jiaotong University, Shanghai 200233, China
  • Online:2012-11-28 Published:2012-11-30
  • Supported by:

    Shanghai Science and Technology Committee Foundation, 054119516

摘要:

目的 研究静脉注射谷氨酰胺(Gln)对脓毒血症大鼠肠淋巴液抑制RAW264.7细胞分泌功能作用的影响。方法 将SD大鼠随机分为假手术组(Sham)、脓毒血症组(CLP)和Gln组,每组10只。取CLP组和Gln组大鼠,以盲肠结扎穿孔法制备脓毒血症模型,术后10 min 内Gln组经尾静脉注射Gln 0.75 g/kg,Sham组和CLP组注射等量生理盐水。各组分别取5只大鼠引流肠淋巴液,另5只于术后6 h时采集血样。将各组淋巴液分别与RAW264.7细胞共孵育4 h,收集培养上清液。ELISA法测定肠淋巴液和细胞培养上清液中肿瘤坏死因子α (TNF-α)和白介素6 (IL-6)的浓度,比色法检测肠淋巴液Gln及血浆D-乳酸浓度。结果 CLP组和Gln组肠淋巴液中TNF-α和IL-6的含量均明显高于Sham组(P<0.05);CLP组细胞培养上清液中TNF-α和IL-6的浓度显著低于Sham组(P<0.05),而Gln组细胞培养上清液中TNF-α浓度显著高于CLP组(P<0.05)。三组肠淋巴液中Gln浓度比较差异均无统计学意义(P>0.05)。Gln组血浆中D-乳酸浓度明显低于CLP组(P<0.05)。结论 脓毒血症大鼠肠淋巴液对LPS激活后的RAW264.7细胞分泌功能有明显抑制作用,Gln能部分减轻该抑制作用。

关键词: 谷氨酰胺, 脓毒血症, 肠淋巴液, 巨噬细胞, 肿瘤坏死因子α, 白介素6

Abstract:

Objective To investigate the effects of intravenous administration of glutamine (Gln) on secretion suppression of RAW 264.7 cells induced by mesenteric lymph in rats with sepsis. Methods SD rats were randomized into sham-operation group (Sham group), sepsis group (CLP group) and Gln group, with 10 rats in each group. Sepsis model was established by cecal ligation and puncture in CLP group and Gln group. Within 10 minutes after model establishment, 0.75 g/kg Gln was administered via single tail-vein injection in Gln group, while the same amount of normal saline was administered in Sham group and CLP group. Mesenteric lymph was obtained from 5 rats in each group, and blood samples were taken from the other rats 6 h after model establishment in each group. RAW264.7 cells were incubated with lymph for 4 h in each group, and the supernatants were harvested. The concentrations of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in mesenteric lymph and supernatants were measured by ELISA, and the concentrations of Gln in mesenteric lymph and D-lactate in plasma were determined by spectrophotometric assay. Results The concentrations of TNF-α and IL-6 in mesenteric lymph in CLP group and GLN group were significantly higher than those in Sham group (P<0.05). The concentrations of TNF-α and IL-6 in supernatants in CLP group were significantly lower than those in sham group (P<0.05), while the concentration of TNF-α in supernatants in Gln group was significantly higher than that in CLP group (P<0.05). There was no significant difference in the concentrations of Gln in mesenteric lymph among three groups (P>0.05). The concentration of D-lactate in plasma in Gln group was significantly lower than that in CLP group (P<0.05). Conclusion Secretion of RAW264.7 cells activated by LPS can be suppressed by mesenteric lymph in rats with sepsis, which can be partially alleviated by Gln treatment.

Key words: glutamine, sepsis, mesenteric lymph, macrophage, tumor necrosis factor-α, interleukin-6