巨噬细胞p38MAPK的激活与钙通道阻滞剂的关系

Relationship of calcium antagonist to the phosphorylated p38 MAPK in LPS-treated M

目的 探讨p38丝裂酶原活化蛋白激酶 (p38MAPK)在炎症反应细胞内的激活与细胞内第二信使—钙离子的关系。方法 分离大鼠腹腔巨噬细胞 (M) ,分为维拉帕米预处理组、SB2 0 35 80预处理组及对照组 ,再予内毒素 (LPS)刺激 ,观察细胞内磷酸化p38的变化及细胞上清液中肿瘤坏死因子 (TNF -α)含量的变化。结果 LPS处理后M中p38磷酸化水平较高 ,而SB2 0 35 80预处理组细胞内p38磷酸化水平明显低于对照组 (P <0 0 1) ,维拉帕米预处理组细胞内p38磷酸化水平与对照组相比无明显差异 (P >0 0 5 )。维拉帕米预处理组和SB2 0 35 80预处理组细胞上清液中TNF -α的含量均明显低于对照组。结论 钙通道阻滞剂维拉帕米可明显减少LPS刺激后MTNF -α的产生 (P <0 0 1) ;维拉帕米预处理组的M经LPS刺激后 ,磷酸化p38仍有较高水平的表达 ,提示细胞内钙离子并未参与p38的磷酸化过程。

Objective To investigate the relationship between phosphorylated p38 in LPS-treated M and calcium antagonist.Methods Rat M were exposed to LPS in the presence of verapamil(specific calcium antagonist) and SB203580(specific antagonist of p38).Cellular protein were extracted and analyzed by Western blot for phosphorylated form of p38.Tumor necrosis factor(TNF)-α were measured by ELISA on the cellular supernatants.Results Compared with control group, cells pretreated with SB203580 led to reduction of LPS-induced p38 phosphorylation, and verapamil had no significant effect on p38 phosphorylation. TNF-α in the pretreatment group of verapamil and SB 203580 was lower than that in control group.Conclusion Although specific calcium antagonist verapamil can significantly reduce the production of TNF-α in the LPS-treated M,there was no significant relationship between[Ca 2+ ] and the phrosphorylation of p38 in LPS-treated M.