Pruimboom W M, van Dijk A P, Tak C J, Bonta I L, Wilson J H, Zijlstra F J
Department of Internal Medicine II, University Hospital Dijkzigt Rotterdam, The Netherlands.
Prostaglandins Leukot Essent Fatty Acids. 1994 Apr;50(4):183-92. doi: 10.1016/0952-3278(94)90143-0.
Ascites is a readily available source of human macrophages (M phi), which can be used to study M phi functions in vitro. We characterized the mediators of inflammation produced by human peritoneal M phi (hp-M phi) obtained from patients with portal hypertension and ascites. The production of the cytokines interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) was found to be lipopolysaccharide (LPS) concentration dependent (0-10 micrograms/ml) with a maximal production at 10 micrograms/ml and also dependent on the time of exposure to the stimulus (0-36 h). IL-1 beta, IL-6 and TNF-alpha production after LPS administration reached a plateau at 24 h. In vitro stimulation for 24 h with LPS does not influence the eicosanoid production from endogenous arachidonate. 13 min of exposure of the cells to the calcium ionophore A23187 gives a significant increase in eicosanoid production from both exogenous and endogenous arachidonate. The main eicosanoids produced are the 5-lipoxgenase products LTB4 and 5-hydroxyeicosatetraenoic acid (HETE). The increase in production of the other eicosanoids is not significant. The eicosanoid production depends on the stimulus concentration. The optimal A23187 concentration is 1 microM. Oxygen radical production was measured in the M phi by a flowcytometric method. The fluorescence intensity of phorbol 12-myristate 13-acetate stimulated and dihydro-rhodamine 123 loaded hp-M phi increases significantly after 15 min. We conclude that LPS stimulation of hp-M phi from liver disease results in similar production of IL-1 beta, IL-6 and TNF-alpha, but that the profile of the eicosanoid production of these M phi stimulated with LPS and A23187 differs from M phi of other origin and species.
腹水是人类巨噬细胞（M phi）的一个容易获取的来源，可用于体外研究M phi的功能。我们对从门静脉高压和腹水患者获得的人腹膜巨噬细胞（hp-M phi）产生的炎症介质进行了表征。发现细胞因子白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）的产生呈脂多糖（LPS）浓度依赖性（0 - 10微克/毫升），在10微克/毫升时产量最高，并且还取决于暴露于刺激物的时间（0 - 36小时）。给予LPS后，IL-1β、IL-6和TNF-α的产生在24小时达到平台期。用LPS体外刺激24小时不影响内源性花生四烯酸的类花生酸产生。细胞暴露于钙离子载体A23187 13分钟会使外源性和内源性花生四烯酸的类花生酸产生显著增加。产生的主要类花生酸是5-脂氧合酶产物白三烯B4和5-羟基二十碳四烯酸（HETE）。其他类花生酸产量的增加不显著。类花生酸的产生取决于刺激物浓度。最佳A23187浓度为1微摩尔。通过流式细胞术方法测量巨噬细胞中的氧自由基产生。佛波醇12-肉豆蔻酸酯13-乙酸酯刺激并加载二氢罗丹明123的hp-M phi在15分钟后荧光强度显著增加。我们得出结论，LPS刺激肝病患者的hp-M phi会导致IL-1β、IL-6和TNF-α产生相似，但用LPS和A23187刺激的这些巨噬细胞的类花生酸产生谱与其他来源和物种的巨噬细胞不同。
