Fidel P L, Romero R, Ramirez M, Cutright J, Edwin S S, LaMarche S, Cotton D B, Mitchell M D
Department of Obstetrics and Gynecology, Wayne State University School of Medicine, Detroit, Michigan.
Am J Reprod Immunol. 1994 Aug;32(1):1-7. doi: 10.1111/j.1600-0897.1994.tb00872.x.
This study was conducted to determine whether (1) conditioned media from unstimulated primary cultures of human amnion, chorion, or decidua contain detectable concentrations of IL-1ra in vitro, and (2) bacterial endotoxin (LPS), tumor necrosis factor-alpha (TNF-alpha), or IL-1-beta (IL-1 beta) stimulate amnion, chorion, or decidua to produce increased amounts of IL-1ra.
Placentae were obtained from women at term with intact membranes before the onset of labor. Amnion, chorion, and decidual cells were isolated by standard procedures and grown to confluence. Cells were then cultured in quadruplicate for 16 h in tissue culture medium supplemented with 10% fetal calf serum or, additionally, with various concentrations of Escherichia coli LPS, TNF-alpha, or IL-1 beta. Culture supernatants were collected, and concentrations of IL-1ra were quantitated by a sensitive and specific enzyme-linked immunosorbent assay for IL-1ra.
Results showed that primary cultures of amnion and chorion from 4 of 9 and decidua from 10 of 12 placentae had detectable rates of production of IL-1ra (ranges: 0.08-6.5, 0.42-12.1, and 1.55-96.5 pg IL-1ra/microgram protein/16 h, respectively). In addition, LPS (10-1,000 ng/ml) and IL-1 beta (0.1-10 ng/ml), but not TNF-alpha (0.01-100 ng/ml), stimulated decidual cells to release/secrete increased amounts of IL-1ra compared with media alone (range: 2.5-400 pg IL-1ra/microgram protein/16 h, P < 0.0001). In contrast, neither LPS, TNF-alpha, or IL-1 beta could stimulate amnion or chorion to release/secrete IL-1ra.
These results indicate (1) that amnion, chorion, and predominantly decidua, can release or secrete IL-1ra in vitro, and (2) that LPS and IL-1 beta can stimulate decidual cells to produce increased amounts of IL-1ra.
本研究旨在确定:(1)来自人羊膜、绒毛膜或蜕膜未受刺激的原代培养物的条件培养基在体外是否含有可检测浓度的IL-1ra;(2)细菌内毒素(LPS)、肿瘤坏死因子-α(TNF-α)或白细胞介素-1β(IL-1β)是否刺激羊膜、绒毛膜或蜕膜产生更多量的IL-1ra。
在临产前从胎膜完整的足月妇女获取胎盘。通过标准程序分离羊膜、绒毛膜和蜕膜细胞并培养至汇合。然后将细胞一式四份培养16小时,培养基为补充有10%胎牛血清的组织培养基,或者另外添加不同浓度的大肠杆菌LPS、TNF-α或IL-1β。收集培养上清液,通过一种针对IL-1ra的灵敏且特异的酶联免疫吸附测定法定量IL-1ra的浓度。
结果显示,9个胎盘中4个的羊膜原代培养物、12个胎盘中10个的绒毛膜原代培养物以及蜕膜原代培养物具有可检测的IL-1ra产生率(范围分别为:0.08 - 6.5、0.42 - 12.1和1.55 - 96.5 pg IL-1ra/μg蛋白质/16小时)。此外,与单独培养基相比,LPS(10 - 1000 ng/ml)和IL-1β(0.1 - 10 ng/ml),而非TNF-α(0.01 - 100 ng/ml),刺激蜕膜细胞释放/分泌更多量的IL-1ra(范围:2.5 - 400 pg IL-1ra/μg蛋白质/16小时,P < 0.0001)。相反,LPS、TNF-α或IL-1β均不能刺激羊膜或绒毛膜释放/分泌IL-1ra。
这些结果表明:(1)羊膜、绒毛膜以及主要是蜕膜在体外可释放或分泌IL-1ra;(2)LPS和IL-1β可刺激蜕膜细胞产生更多量的IL-1ra。