Department of Obstetrics and Gynecology, Rostock University Medical Center, Suedring 81, 18059, Rostock, Germany.
Department of Obstetrics and Gynecology, Rostock University Medical Center, Suedring 81, 18059, Rostock, Germany.
Placenta. 2020 Jan 15;90:109-117. doi: 10.1016/j.placenta.2019.12.018. Epub 2019 Dec 24.
Functional disorders of the villous trophoblast may result in preeclampsia through the release of endothelial activating substances. Progranulin is an anti-inflammatory, pro-angiogenic cytokine with TNF-α antagonizing activity. The trophoblastic expression of progranulin is increased during preeclampsia. The aim of the study was to investigate the impact of placental progranulin synthesis on endothelial cell activation.
Placental progranulin expression was modified by transduction of an adenoviral vector. Primary isolated human umbilical venous endothelial cells (HUVECs) were incubated with conditioned medium of first trimester placental explants. Functional studies on HUVECs included assays for proliferation, viability, cytotoxicity and analyzes of Intercellular adhesion molecule-1 (ICAM-1) and E-selectin expression.
Placental progranulin expression was more than 10-fold higher by using an adenoviral-mediated overexpression system (Ad.PGRN) compared to control vector (Ad.CTRL) and untreated controls. Incubation of HUVECs with conditioned placental medium revealed a dose-dependent increase of cytotoxicity, reduced cell proliferation and viability and resulted in an increase of ICAM-1 and E-selectin expression. Overexpression of progranulin (Ad.PGRN) antagonized the ICAM-1 expression induced by conditioned medium. However progranulin did not influence the effects on cell proliferation, viability, cytotoxicity and E-selectin expression in HUVECs.
Regulation of gene expression in human placental explants is possible by usage of an adenoviral vector system. The increase of endothelial ICAM-1 expression following the incubation with placental conditioned medium was partly reversed by overexpression of placental progranulin. It is suggested that up-regulation of the placental progranulin expression is an endogenous anti-inflammatory mechanism that partially antagonizes the endothelial cell activation during preeclampsia.
绒毛滋养层的功能障碍可能通过释放内皮激活物质导致子痫前期。颗粒蛋白前体是一种具有 TNF-α拮抗活性的抗炎、促血管生成细胞因子。颗粒蛋白前体在子痫前期时的胎盘表达增加。本研究旨在探讨胎盘颗粒蛋白前体合成对内皮细胞激活的影响。
通过转导腺病毒载体来改变胎盘颗粒蛋白前体的表达。原代分离的人脐静脉内皮细胞（HUVEC）与第一孕期胎盘外植体的条件培养基孵育。HUVEC 的功能研究包括增殖、活力、细胞毒性测定以及细胞间黏附分子-1（ICAM-1）和 E-选择素表达分析。
与对照载体（Ad.CTRL）和未处理对照组相比，使用腺病毒介导的过表达系统（Ad.PGRN）使胎盘颗粒蛋白前体的表达增加了 10 多倍。孵育 HUVEC 的条件胎盘培养基显示细胞毒性呈剂量依赖性增加，细胞增殖和活力降低，并导致 ICAM-1 和 E-选择素表达增加。颗粒蛋白前体的过表达（Ad.PGRN）拮抗了由条件培养基诱导的 ICAM-1 表达。然而，颗粒蛋白前体对 HUVEC 中的细胞增殖、活力、细胞毒性和 E-选择素表达没有影响。
使用腺病毒载体系统可以调节人胎盘外植体中的基因表达。孵育胎盘条件培养基后内皮细胞 ICAM-1 表达增加，部分被胎盘颗粒蛋白前体过表达逆转。提示上调胎盘颗粒蛋白前体的表达是一种内源性抗炎机制，部分拮抗子痫前期内皮细胞激活。
