咯利普兰通过MAPK/ERK信号通路抑制中性粒细胞的黏附  

Rolipram inhibits neutrophils adhesion by MAPK and ERK signaling pathway

作　　者：孔学礼 王亚楠 丛心宇 李佳 陈武 姜代勋 KONG Xueli;WANG Yanan;CONG Xinyu;LI Jia;CHEN Wu;JIANG Daixun(Animal Science and Technology College/Beijing Key Laboratory of Traditional Chinese Veterinary Medicine,Beijing University of Agriculture,Beijing 102206,China)

机构地区：[1]北京农学院动物科学技术学院/兽医学(中医药)北京市重点实验室,北京102206

出　　处：《北京农学院学报》2018年第2期58-63,共6页Journal of Beijing University of Agriculture

基　　金：北京市自然科学基金资助项目(6152003/6112007)

摘　　要：【目的】研究磷酸二酯酶4(PDE4)特异性抑制剂咯利普兰抑制中性粒细胞黏附的有关分子机制,明确其抗炎的主要信号通路,为PDE4选择性中药抑制剂的药理研究提供参考。【方法】猪中性粒细胞的提取采用密度梯度离心法,中性粒细胞与猪内皮细胞的黏附采用虎红比色法,中性粒细胞黏附分LFA-1和Mac-1表达量的检测采用流式细胞技术,中性粒细胞p38MAPK、ERK等磷酸化活性检测采用western blot法。【结果】咯利普兰可显著抑制fMLP刺激的中性粒细胞和内皮细胞的黏附(P<0.05),极显著抑制fMLP刺激的中性粒细胞LFA-1和Mac-1的表达(P<0.01),阻断fMLP刺激的中性粒细胞ERK磷酸化活性(P<0.01),而对p38MAPK磷酸化活性无显著影响。【结论】咯利普兰可抑制中性粒细胞和内皮细胞黏附,其机制与阻断中性粒细胞ERK磷酸化进而抑制LFA-1和Mac-1表达有关。【Objective】To study the molecular mechanism of rolipram,a specific inhibitor of phosphodiesterase type 4(PDE4),on the inhibition of neutrophils adhesion,elucidate the main pathway of anti-inflammation,and provide a reference for the pharmacological study of PDE4 selective herbal inhibitors.【Methods】Density gradient centrifugation was used to extract porcine neutrophils,Tiger red colorimetric method was used to detect the adhesion of neutrophils to porcine endothelial cells,the adhesion molecules,LFA-1 and Mac-1,of neutrophils were detected by flow cytometry,and the phosphorylation of p38MAPK and ERK in neutrophils was detected by Western blot.【Results】Rolipram could significantly inhibit fMLP-stimulated adhesion of neutrophils and endothelial cells(P<0.05),significantly inhibit fMLP-stimulated expression of LFA-1 and Mac-1 on neutrophils(P<0.01),block fMLP-stimulated ERK phosphorylation activity in neutrophils(P<0.01),but could not show any significant effect on p38MAPK phosphorylation.【Conclusion】Rolipram can inhibit the adhesion of neutrophils and endothelial cells,and its mechanism maybe relate to blocking ERK phosphorylation and inhibiting LFA-1 and Mac-1 expression of neutrophils.

关 键 词：ERK p38MAPK 黏附分子 LFA-1 MAC-1 中性粒细胞 

分 类 号：S859.7[农业科学—临床兽医学]