Pollard J K, Mitchell M D
Department of Obstetrics and Gynecology, Medical Center Hospital of Vermont, Burlington.
Prostaglandins. 1993 Dec;46(6):499-510. doi: 10.1016/0090-6980(93)90069-j.
The purpose of this study was to determine how tumor necrosis factor alpha (TNF alpha) stimulates prostaglandin E2 production in human amnion. Amnion cells were isolated from term placentae and grown to confluence in culture. Incubations were conducted in quadruplicate wells for 16 hours with TNF alpha and protein synthesis inhibitors cycloheximide and actinomycin D, or arachidonic acid, acetylsalicylic acid (ASA), or staurosporine or H7 which inhibit protein kinase C activity. Prostaglandin E2 (PGE2) was measured by radioimmunoassay and cellular protein determined. The stimulatory action of TNF alpha on amnion PGE2 production was blocked by protein synthesis inhibitors, and the addition of arachidonic acid always enhanced the stimulatory properties of TNF alpha. TNF alpha consistently induced more rapid recovery from ASA treatment, and protein kinase C inhibition attenuated the stimulatory effects of TNF alpha. These results suggest that the stimulatory action of TNF alpha on amnion PGE2 production is likely at the level of induction of fatty acid cyclooxygenase activity and is partially dependent upon activation of protein kinase C.
本研究的目的是确定肿瘤坏死因子α（TNFα）如何刺激人羊膜中前列腺素E2的产生。从足月胎盘分离羊膜细胞，并在培养中生长至汇合。用TNFα和蛋白质合成抑制剂环己酰亚胺和放线菌素D，或花生四烯酸、乙酰水杨酸（ASA），或抑制蛋白激酶C活性的星形孢菌素或H7在一式四份的孔中进行孵育16小时。通过放射免疫测定法测量前列腺素E2（PGE2）并测定细胞蛋白。蛋白质合成抑制剂阻断了TNFα对羊膜PGE2产生的刺激作用，添加花生四烯酸总是增强TNFα的刺激特性。TNFα始终能使细胞从ASA处理中更快恢复，蛋白激酶C抑制减弱了TNFα的刺激作用。这些结果表明，TNFα对羊膜PGE2产生的刺激作用可能在脂肪酸环氧化酶活性诱导水平，并且部分依赖于蛋白激酶C的激活。
