Schmidt J, Hatzelmann A, Fleissner S, Heimann-Weitschat I, Lindstaedt R, Szelenyi I
ASTA Medica AG, Department of Pharmacology, Frankfurt/Main, Germany.
Immunopharmacology. 1995 Sep;30(3):191-8. doi: 10.1016/0162-3109(95)00022-l.
The effect of various phosphodiesterase (PDE) inhibitors on anti-CD3 induced interleukin-(IL)-4 and IL-5 production of the murine T helper cell clone of type 2 phenotype D10.G4.1 (D10) has been investigated in vitro. D10 cells were incubated in the presence of drugs and anti-CD3 mAb for 16 h before measurement of cytokines in the cell supernatants by ELISA. Whereas all PDE inhibitors tested exerted minimal effects on anti-CD3 induced IL-4 production, a marked increase in IL-5 production by the non-selective PDE inhibitors IBMX, theophylline and enprofylline was observed. The action of these non-selective PDE inhibitors was mimicked by the PDE IV-selective inhibitor rolipram and in part by the PDE III-selective inhibitors motapizone and milrinone, whereas the PDE V-selective inhibitor zaprinast was inactive. Rolipram and motapizone enhanced IL-5 production in a synergistic fashion. In support of the functional importance of PDE III and IV for IL-5 synthesis in intact murine D10 cells, we have found PDE III and IV to be the predominant isoenzyme activities in corresponding cell lysates. The stimulatory effect of rolipram on IL-5 production was almost totally reversed by the protein kinase A inhibitor KT-5720. In addition, the membrane-permeable cAMP analogue 8-bromo-cAMP mimicked the stimulatory effect of PDE inhibitors on IL-5 production while leaving IL-4 levels unaffected. Both results support the view that the action of the PDE inhibitors on murine D10 cells is mediated via an elevation of intracellular cAMP.
已在体外研究了各种磷酸二酯酶（PDE）抑制剂对由抗CD3诱导的2型表型小鼠T辅助细胞克隆D10.G4.1（D10）产生白细胞介素-（IL）-4和IL-5的影响。在通过酶联免疫吸附测定法（ELISA）测量细胞上清液中的细胞因子之前，将D10细胞与药物和抗CD3单克隆抗体一起孵育16小时。尽管所测试的所有PDE抑制剂对抗CD3诱导的IL-4产生的影响极小，但观察到非选择性PDE抑制剂异丁基甲基黄嘌呤（IBMX）、茶碱和恩丙茶碱可显著增加IL-5的产生。PDE IV选择性抑制剂咯利普兰以及部分PDE III选择性抑制剂莫他匹宗和米力农可模拟这些非选择性PDE抑制剂的作用，而PDE V选择性抑制剂扎普司特则无活性。咯利普兰和莫他匹宗以协同方式增强IL-5的产生。为了支持PDE III和IV对完整小鼠D10细胞中IL-5合成的功能重要性，我们发现PDE III和IV是相应细胞裂解物中的主要同工酶活性。蛋白激酶A抑制剂KT-5720几乎完全逆转了咯利普兰对IL-5产生的刺激作用。此外，膜通透性cAMP类似物8-溴-cAMP模拟了PDE抑制剂对IL-5产生的刺激作用，而对IL-4水平无影响。这两个结果均支持以下观点：PDE抑制剂对小鼠D10细胞的作用是通过细胞内cAMP升高介导的。
