Hulkower K I, Otis E R, Li J, Ennis B W, Cugier D J, Bell R L, Carter G W, Glaser K B
Immunosciences Research Area, Abbott Laboratories, Abbott Park, Illinois 60064-3500, USA.
J Pharmacol Exp Ther. 1997 Feb;280(2):1065-74.
These studies examined the signal transduction mechanisms by which prostaglandin (PG) E2 production can occur in human amnionic WISH cells in response to the stimuli okadaic acid, interleukin (IL)-1beta, tumor necrosis factor (TNF)-alpha, phorbol-12-myristate-13-acetate (PMA) or combinations of PMA with IL-1beta or TNF-alpha. We also investigated whether WISH cells are capable of producing TNF-alpha or IL-1beta in response to stimulation, because these cytokines can be produced in an autocrine fashion to perpetuate an inflammatory response. Our data indicate that the magnitude of PGE2 production induced by a given stimulus correlated temporally with the level of PGH synthase-2 (PGHS-2) protein. PMA or IL-1beta induced PGE2 production 2 to 4 hr after treatment, whereas the combination of these agents produced the most rapid induction 2 hr after treatment. Only okadaic acid induced the production of both PGE2 and TNF-alpha, after a lag of 12 to 18 hr. PGE2 production by all stimuli was inhibited by dexamethasone, the IL-1 receptor antagonist (IL-1ra), the specific PGHS-2 inhibitor NS-398 and the protein kinase inhibitor staurosporin. In contrast, TNF-alpha production in response to okadaic acid was inhibited by the TNF-converting enzyme inhibitor GI 129471 and staurosporin but was unaffected by either IL-1ra, dexamethasone or NS-398. We conclude that WISH cells are capable of producing bioactive proinflammatory mediators such as TNF-alpha and PGE2 through separable intracellular signal transduction mechanisms. The ability of IL-1ra to reduce PGE2 production caused by all stimuli used suggests an autocrine role for IL-1 in PGHS-2 induction in these cells.
这些研究检测了前列腺素（PG）E2 在人羊膜 WISH 细胞中因冈田酸、白细胞介素（IL）-1β、肿瘤坏死因子（TNF）-α、佛波酯-12-肉豆蔻酸酯-13-乙酸酯（PMA）或 PMA 与 IL-1β 或 TNF-α 的组合刺激而产生的信号转导机制。我们还研究了 WISH 细胞是否能够在受到刺激时产生 TNF-α 或 IL-1β，因为这些细胞因子可以以自分泌方式产生，从而使炎症反应持续存在。我们的数据表明，给定刺激诱导产生的 PGE2 量在时间上与前列腺素 H 合酶-2（PGHS-2）蛋白水平相关。PMA 或 IL-1β 在处理后 2 至 4 小时诱导 PGE2 产生，而这些试剂的组合在处理后 2 小时产生最快的诱导。仅冈田酸在滞后 12 至 18 小时后诱导 PGE2 和 TNF-α 产生。所有刺激引起的 PGE2 产生均受到地塞米松、IL-1 受体拮抗剂（IL-1ra）、特异性 PGHS-2 抑制剂 NS-398 和蛋白激酶抑制剂星形孢菌素的抑制。相反，对冈田酸产生的 TNF-α 反应受到 TNF 转换酶抑制剂 GI 129471 和星形孢菌素的抑制，但不受 IL-1ra、地塞米松或 NS-398 的影响。我们得出结论，WISH 细胞能够通过可分离的细胞内信号转导机制产生生物活性促炎介质，如 TNF-α 和 PGE2。IL-1ra 降低所有所用刺激引起的 PGE2 产生的能力表明 IL-1 在这些细胞中 PGHS-2 诱导中具有自分泌作用。
