摘要
目的观察脓毒症大鼠血清对RAW264．7细胞的免疫刺激作用及谷氨酰胺（Gin）的影响，并探讨其保护作用的机制。方法SD大鼠18只，随机分为假手术组（sham组）、脓毒症组、Gin处理组（GLN组）。采用盲肠结扎穿孔术（CLP）制备大鼠脓毒症模型，术后即刻从尾静脉注射Gln0．75g／kg（GLN组）或等量生理盐水（sham和脓毒症组）。3组均于术后6h采血并取肺组织，并将血清与RAW264．7细胞孵育4h后收集上清液。鲎试剂法定量测定血浆内毒素浓度；比色法检测血浆D-乳酸和血清Gin水平；酶联免疫吸附法（ELISA）测定肺组织及血清作用于RAW264．7细胞后上清液中肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）水平。结果3组血清Gln浓度无明显差异。GLN组血浆D-乳酸和内毒素水平明显低于脓毒症组[D-乳酸：（2．04±0．13）mg／L比（5．08±0．47）mg／L；内毒素：（0．011±0．003）mg／L比（5．085±1．647）mg／L，P均〈0．01]。脓毒症组血清刺激内毒素激活的RAW264．7细胞释放炎症因子明显高于sham组，GLN组明显低于脓毒症组（P〈0．05或P〈0．01），而GLN组与sham组比较差异无统计学意义。GLN组肺组织匀浆中TNF—α浓度明显低于脓毒症组[（31．01±14．23）μg／L比（80．18±26．27）μg／L，P〈0．01]。结论脓毒症大鼠血清对巨噬细胞释放炎症因子有明显的促进作用，而Gin可抑制脓毒症血清的该作用，这可能与Gin改变肠道通透性有关。
Objective To investigate the effects of serum in rats with sepsis on RAW264.7 macrophage immunostimulation, the influence of glutamine （Gln） on the serum and the protective effects of Gln on the organism. Methods Eighteen Sprague-Dawley （SD） rats were randomly divided into three groups： sham operation group （sham group）, sepsis group and Gin-treatment group （GLN group）. The septic model was established by cecal ligation and puncture （CLP）. After operation, Gln 0.75 g/kg （GLN group） or saline （sham and CLP groups） was immediately injected into a tail vein. Serum and lung tissue were collected at 6 post-operative hours in these three groups, and the supernatant was collected after RAW264.7 macrophages were incubated in the serum for 4 hours. The concentration of endotoxin （LPS） in plasma was detected by limulus test ; the levels of Dqactate in plasma and Gln in serum were detected by spectrophotometric assay. The level of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in lung tissue and the supernatant collected as the method above was detected by enzyme linked immunosorbent assay （ELISA）. Results There was no difference of Gln concentration in serum in the three groups. In GLN group, the plasma concentrations of D-lactate and LPS were significantly lower than those in CLP group [D-lactate： （2.04±0.13） mg/L vs. （5.08±0.47） mg/L, LPS： （0.011±0.003） mg/L vs. （5.085±1.647） mg/L, both P〈0.01]. The serum of CLP group stimulated the RAW264.7 macrophages activated by LPS to release higher cytokines than those in sham group, while in the GLN group, the cytokines released were remarkably lower than those in CLP group （P〈0.05 or P〈0.01）, but there was no difference between GLN group and sham group. The concentration of TNF-α in lung tissue homogenate of GLN group was significantly lower than that in CLP group [（31. 01±＋14. 23） μg/L vs. （80. 18±26.27） μg/L, P〈0.01]. Conclusion The serum of rats with sepsis can apparently improve the effect of macrophage to release cytokines, and Gln may inhibit this serum action. The mechanism is probably related to the change of intestinal permeability by Gln.
出处
《中国中西医结合急救杂志》
CAS
北大核心
2009年第1期38-41,共4页
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care
基金
上海市科委科研计划项目（054119516）
