Gallery E D, Rowe J, Campbell S
Department of Renal Medicine, Sydney University, Royal North Shore Hospital, St Leonards, NSW, Australia.
Prostaglandins Leukot Essent Fatty Acids. 1996 Jun;54(6):411-8. doi: 10.1016/s0952-3278(96)90024-1.
Endothelial cells isolated from umbilical veins (HUVEC) and from decidual biopsies collected at caesarean section delivery (DEC) from both normal (N DEC) and pre-eclamptic (PE DEC) women, were maintained in culture until passage 2, when the effect on growth of removing heparin/ECGS (endothelial cell growth supplement) from the culture medium was assessed, and the effects of heparin-free incubation and of the Ca2+ ionophore A23187 on endothelin-1, prostacyclin and prostaglandin E2 secretion over a 24 h period were examined. Cell growth slowed significantly in all three cell types in the absence of heparin/ECGS, and cell death occurred in 1/3 samples of HUVEC, 4/6 of N DEC, but 0/2 of PE DEC over 4 days. During the 24 h incubation for prostaglandin in medium without these growth factors, there was further cell death in N DEC. The addition of A23187 to this stress led to a reduction in cell number in both N DEC and HUVEC, and to a lesser extent in PE DEC. Prostaglandin and endothelin-1 levels were higher in the absence of heparin/ECGS in all cell types There was significant suppression of endothelin-1 secretion at 24 h incubation, and stimulation of prostaglandin secretion by A23187. Incubation without heparin/ECGS magnified the effect of A23187 on prostaglandin secretion, although the proportional change was similar if compared to controls without heparin/ECGS. Withdrawal of heparin/ECGS from the medium altered the balance of PGE2/PGI2 secretion by HUVEC, but not DEC. Endothelial cells require the presence of heparin/ECGS for optimum growth and viability, and N DEC are particularly dependent on these growth factors. PE DEC appear relatively 'hardy' in this regard. The addition of a further potentially toxic stimulus may result in cell death, and experiments to be conducted in limited medium must take this into account. There are both qualitative and quantitative differences in the effects of these stimuli on secretion of vasoactive substances, between decidual and umbilical vein endothelial cells.
从脐静脉分离的内皮细胞（人脐静脉内皮细胞）以及从剖宫产分娩时收集的蜕膜活检组织中分离的内皮细胞（蜕膜细胞），这些蜕膜活检组织取自正常（正常蜕膜细胞）和先兆子痫（先兆子痫蜕膜细胞）女性。将这些细胞培养至第2代，此时评估从培养基中去除肝素/内皮细胞生长补充剂（ECGS）对细胞生长的影响，并检测无肝素培养以及钙离子载体A23187在24小时内对内皮素-1、前列环素和前列腺素E2分泌的影响。在没有肝素/ECGS的情况下，所有三种细胞类型的细胞生长均显著减缓，并且在4天内，人脐静脉内皮细胞的1/3样本、正常蜕膜细胞的4/6样本发生细胞死亡，但先兆子痫蜕膜细胞的2/0样本未发生细胞死亡。在没有这些生长因子的培养基中进行24小时前列腺素孵育期间，正常蜕膜细胞进一步发生细胞死亡。向这种应激状态下添加A23187导致正常蜕膜细胞和人脐静脉内皮细胞的细胞数量减少，先兆子痫蜕膜细胞减少程度较小。在所有细胞类型中，没有肝素/ECGS时前列腺素和内皮素-1水平较高。在24小时孵育时内皮素-1分泌受到显著抑制，A23187刺激前列腺素分泌。无肝素/ECGS孵育放大了A23187对前列腺素分泌的影响，尽管与没有肝素/ECGS的对照相比，比例变化相似。从培养基中去除肝素/ECGS改变了人脐静脉内皮细胞中前列腺素E2/前列环素分泌的平衡，但未改变蜕膜细胞的平衡。内皮细胞需要肝素/ECGS的存在以实现最佳生长和活力，正常蜕膜细胞尤其依赖这些生长因子。在这方面，先兆子痫蜕膜细胞似乎相对“耐寒”。添加另一种潜在的毒性刺激可能导致细胞死亡，在有限培养基中进行的实验必须考虑到这一点。这些刺激对血管活性物质分泌的影响在蜕膜和脐静脉内皮细胞之间存在质和量的差异。
