Vascular Immunology Research Laboratory, The Heart Research Institute, University of Sydney, Australia.
Vascular Immunology Research Laboratory, The Heart Research Institute, University of Sydney, Australia; School of Medicine, Western Sydney University, Sydney, Australia; Renal Unit, Liverpool Hospital, Sydney, Australia.
Pregnancy Hypertens. 2020 Jan;19:138-142. doi: 10.1016/j.preghy.2020.01.004. Epub 2020 Jan 14.
Integrins are cell adhesion receptors that participated in endovascular invasion by cytotrophoblasts in preeclampsia. This study aimed to investigate the effect of calcium on cellular pathways influencing the trophoblast integration into endothelial cellular networks in vitro.
Red fluorescent-labelled human uterine myometrial microvascular endothelial cells (UtMVECs) were seeded on Matrigel. Green fluorescent-labelled HTR-8/SVneo trophoblast cells were then co-cultured with endothelial cells in different concentrations of calcium for 24 h.
The calcium effects on HTR-8/SVneo cell integration were quantified by Image J. Quantitative PCR was performed to measure mRNA expression of integrins α1, α5, α6, β1 and β4. The concentrations of interleukin IL-6, matrix metalloproteinase-2 (MMP-2), MMP-9, PlGF and sFlt-1 in the conditioned medium were measured by ELISA while levels of cytokines IL-1β, IL-8, IL-10, TNF-α and INF-γ were assessed by magnetic Luminex assays™.
Both calcium depletion (0.4 mM) and low calcium (1.8 mM) groups demonstrated inhibited integration of trophoblast cells into endothelial cellular networks, compared with the normal calcium group (2.4 mM). The IL-6 production was reduced from conditioned media in both calcium depletion and low calcium groups. In calcium depletion group, mRNA expression of integrin α5 and β4 in trophoblasts was increased while integrin α1 was decreased.
The in vitro trophoblast cell integration into endothelial cellular networks could be modified by altering media calcium through integrin switch away from integrins α5 and β4 and towards integrin α1 which may be required for healthy early trophoblast integration.
整合素是细胞黏附受体，参与子痫前期细胞滋养层的血管内侵袭。本研究旨在探讨钙对影响滋养细胞整合入内皮细胞网络的细胞通路的影响。
将红色荧光标记的人子宫平滑肌微血管内皮细胞（UtMVECs）接种在 Matrigel 上。然后，将绿色荧光标记的 HTR-8/SVneo 滋养细胞与内皮细胞在不同浓度的钙中共同培养 24 小时。
通过 Image J 定量滋养细胞 HTR-8/SVneo 细胞整合的钙效应。采用定量 PCR 测量整合素 α1、α5、α6、β1 和 β4 的 mRNA 表达。通过 ELISA 测量条件培养基中白细胞介素（IL）-6、基质金属蛋白酶-2（MMP-2）、MMP-9、胎盘生长因子（PlGF）和可溶性血管内皮生长因子受体 1（sFlt-1）的浓度，通过磁性 Luminex 测定法评估细胞因子 IL-1β、IL-8、IL-10、TNF-α 和 INF-γ 的水平。
与正常钙组（2.4 mM）相比，钙耗竭（0.4 mM）和低钙（1.8 mM）组均显示滋养细胞整合入内皮细胞网络受到抑制。IL-6 的产生在钙耗竭和低钙组的条件培养基中均减少。在钙耗竭组中，滋养细胞中整合素 α5 和 β4 的 mRNA 表达增加，而整合素 α1 减少。
通过改变培养基中的钙，体外滋养细胞整合入内皮细胞网络可以发生改变，通过整合素开关从整合素 α5 和 β4 转向整合素 α1，这可能是健康早期滋养细胞整合所必需的。
