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目的探讨谷氨酰胺(Gln)对内毒素诱导幼鼠脑组织核因子-κB(NF-κB)和热休克蛋白70(HSP70)表达的影响。方法将10日龄Wistar大鼠随机分为内毒素组(LPS组),9 g/L盐水对照组(NS组)和Gln组。LPS组大鼠腹腔注射LPS 5 mg/kg,制备内毒素血症大鼠模型;Gln组于腹腔注射LPS前1 h腹腔注射Gln 1.346 g/kg;对照组腹腔注射同量9 g/L盐水。免疫组织化学方法检测各组幼鼠大脑皮层NF-κB和HSP70的定位表达,Western blot法检测其脑组织HSP70水平。采用SPSS 12.0软件进行统计学分析。结果在腹腔注射LPS后6 h,LPS组幼鼠大脑皮层神经元NF-κB阳性染色定位于细胞核,LPS注射后2 h,Gln组幼鼠NF-κB神经元细胞核阳性染色被明显抑制。免疫组织化学方法检测LPS注射后6 h LPS组幼鼠大脑皮层神经元细胞核HSP70阳性染色明显减弱,免疫印迹分析显示蛋白表达量明显降低,以24 h为显著。LPS组幼鼠LPS注射后2、6 h,其脑组织HSP70表达量接近于NS组,Gln组LPS注射后2 h皮层神经元细胞核HSP70阳性染色信号增强;LPS注射后2、6、12、24 h Gln组HSP70表达明显增加,与LPS组比较均有显著差异(Pa<0.01)。结论Gln可上调内毒素诱导的幼鼠脑组织HSP70表达,并降低NF-κB活化,调节脑组织局部的免疫防御反应。
Objective To investigate the effects of glutamine on the expression of nuclear factor-κB (NF-κB) and heat shock protein 70 (HSP70) in brain of immature rats induced by endotoxin. Methods 10-day-old Wistar rats were randomly divided into LPS group, 9 g / L saline control group (NS group) and Gln group. Rats in LPS group were injected intraperitoneally with LPS 5 mg / kg to prepare rat model of endotoxemia. Gln group was injected intraperitoneally with Gln 1.346 g / kg 1 h before intraperitoneal injection of LPS, while control group received intraperitoneal injection of 9 g / L saline. Immunohistochemistry was used to detect the localization of NF-κB and HSP70 in the cerebral cortex of each group. Western blot was used to detect the level of HSP70 in brain tissue. SPSS 12.0 software was used for statistical analysis. Results The positive staining of NF-κB in the cerebral cortex neurons of LPS group was located in the nucleus 6 h after intraperitoneal injection of LPS. The positive staining of NF-κB neurons in the young Gn group was significantly inhibited 2 h after LPS injection. Immunohistochemistry showed that HSP70 positive staining of neurons in cerebral cortex of LPS group was significantly weakened 6 h after LPS injection, and Western blot analysis showed that the expression of HSP70 protein was significantly decreased at 24 h. At 2 and 6 h after LPS injection, the expression of HSP70 in brain tissue of LPS group was similar to that of NS group. The signal of HSP70 positive staining in nucleus of cortex neurons of LPS group was enhanced 2 h after LPS injection. At 2, 6, 12 and 24 h Gln group HSP70 expression was significantly increased, compared with LPS group were significantly different (Pa <0.01). Conclusion Gln can up-regulate endotoxin-induced HSP70 expression in brain tissue of young rats, decrease the activation of NF-κB and regulate the local immune defense response in brain tissue.