阻断p38MAPK信号通路降低脑死亡大鼠肾脏免疫原性

Blockade of the p38 MAPK signaling pathway reduces the immunogenicity of kidneys from brain dead rats

目的探讨降低脑死亡大鼠肾脏免疫原性的有效途径。方法雄性Wistar大鼠30只，体质量180～200g，随机均分3组，脑死亡＋SB203580组：诱导大鼠脑死亡时，静脉注射SB203580（1mg／100g·Wt）；脑死亡组：诱导大鼠脑死亡。脑死亡后机械呼吸6h，如大鼠血压〉80mm Hg（1mmHg=0．133kPa），取肾脏；对照组：正常大鼠麻醉后取肾脏。逆转录-聚合酶链反应（RT—PCR）检测肾脏肿瘤坏死因子（TNF）-α和白细胞介素（IL）-1β mRNA表达，Western blot检测肾脏磷酸化p38丝裂原活化蛋白激酶（p38MAPK）以及TNF-α和IL-1β蛋白表达。结果肾脏TNF-α和IL-1β mRNA和蛋白表达，以及磷酸化p38MAPK蛋白表达，脑死亡组比对照组显著增加（P〈0．01）；脑死亡＋SB203580组比脑死亡组显著下降（P〈0．05），但比对照组明显增加（P〈0．01）。结论SB203580能阻断p38MAPK信号通路，减少脑死亡大鼠肾脏磷酸化p38MAPK和促炎细胞因子表达，可望成为降低其免疫原性的一条有效途径。

Objective To discuss an efficient way which reduces the immunogenicity of kidneys from brain dead （BD） rats. Methods Thirty male Wistar rats,weighing from 180 to 200 g,were randomly and equally divided into 3 groups：BD ＋ SB203580 group：SB203580 （1 mg/100 g · Wt） was given intravenously into the rats as BD was induced in the rats;BD group：the rat BD was induced, and 6 h after onset of BD,if the mean blood pressure of the rats was more than 80 mmHg, the kidney from the BD rats was harvested;Control group：normal rats whose kidneys were harvested under the condition of anesthesia. RTPCR was used to detect the expression of TNF-α and IL-1β mRNA. Western blot was applied to detect the expression of both TNF-α and IL-1β proteins,and the phosphorylated p38 MAPK proteins. Results The expression levels of both TNF-α and IL-1β mRNA and proteins, and the phosphorylated p38 MAPK proteins were significantly increased in BD group compared to control group （P 〈 0.01 ）, and those in BD ＋ SB203580 group were significantly decreased compared to BD group （ P 〈 0.05 ） , however, those were significantly increased compared to control group （ P 〈 0. 01 ）. Conclusion SB203580 could inhibit p38 MAPK signaling pathway, decrease the expressions of both phosphorylated p38 MAPK proteins and pro-inflammatory cytokines in kidneys from the BD donor rats, and reduce the immunogen city of the kidneys.