Department of Obstetrics and Gynecology, Scott and White Healthcare/Texas A&M Health Science Center College of Medicine, Temple, TX.
Prehealth Studies, Baylor University, Waco, TX.
Am J Obstet Gynecol. 2014 Nov;211(5):541.e1-8. doi: 10.1016/j.ajog.2014.04.033. Epub 2014 May 1.
Diabetes mellitus is a risk factor for preeclampsia. Cytotrophoblast (CTB) invasion is facilitated from the conversion of plasminogen to plasmin by urokinase plasminogen activator (uPA), regulated by plasminogen activator inhibitor 1 (PAI-1), and may be inhibited in preeclampsia. This study assessed signaling mechanisms of hyperglycemia-induced CTB dysfunction.
Human CTBs were treated with 45, 135, 225, 495, or 945 mg/dL glucose for 48 hours. Some cells were pretreated with a p38 inhibitor (SB203580) or a peroxisome proliferator-activated receptor-gamma (PPAR-γ) ligand (rosiglitazone). Expression of uPA, PAI-1, and PPAR-γ levels and p38 mitogen-activated protein kinase phosphorylation were measured by Western blot in cell lysates. Messenger ribonucleic acid of uPA and PAI-1 was measured by quantitative polymerase chain reaction. Levels of interleukin-6, angiogenic (vascular endothelial growth factor [VEGF], placenta growth factor [PlGF]) and antiangiogenic factors (soluble fms-like tyrosine kinase-1 [sFlt-1], soluble endoglin [sEng]) were measured in the media by enzyme-linked immunosorbent assay kits. Statistical comparisons were performed using analysis of variance with a Duncan's post-hoc test.
Both uPA and PAI-1 protein and messenger ribonucleic acid were down-regulated (P < .05) in CTBs treated with 135 mg/dL glucose or greater compared with basal (45 mg/dL). The sEng, sFlt-1, and interleukin-6 were up-regulated, whereas the VEGF and PlGF were down-regulated by 135 mg/dL glucose or greater. p38 phosphorylation and PPAR-γ were up-regulated (P < .05) in hyperglycemia-treated CTBs. The SB203580 or rosiglitazone pretreatment showed an attenuation of glucose-induced down-regulation of uPA and PAI-1.
Hyperglycemia disrupts the invasive profile of CTB by decreasing uPA and PAI-1 expression; down-regulating VEGF and PlGF; and up-regulating sEng, sFlt-1, and interleukin-6. Attenuation of CTB dysfunction by SB203580 or rosiglitazone pretreatment suggests the involvement of stress signaling.
糖尿病是子痫前期的一个危险因素。尿激酶型纤溶酶原激活物（uPA）可促进滋养细胞（CTB）侵袭，纤溶酶原激活物抑制剂 1（PAI-1）调节其转化，子痫前期可能会抑制其转化。本研究评估了高血糖诱导 CTB 功能障碍的信号转导机制。
将人 CTB 用 45、135、225、495 或 945 mg/dL 葡萄糖处理 48 小时。部分细胞用 p38 抑制剂（SB203580）或过氧化物酶体增殖物激活受体-γ（PPAR-γ）配体（罗格列酮）预处理。用 Western blot 法检测细胞裂解物中 uPA、PAI-1 和 PPAR-γ的表达和 p38 丝裂原激活蛋白激酶磷酸化。用定量聚合酶链反应法检测 uPA 和 PAI-1 的信使核糖核酸。用酶联免疫吸附试验试剂盒检测培养基中白细胞介素 6、血管生成（血管内皮生长因子[VEGF]、胎盘生长因子[PlGF]）和抗血管生成因子（可溶性 fms 样酪氨酸激酶-1[sFlt-1]、可溶性内皮糖蛋白[sEng]）的水平。采用方差分析和 Duncan 后测检验进行统计学比较。
与基础（45 mg/dL）相比，135 mg/dL 或更高浓度的葡萄糖处理的 CTB 中 uPA 和 PAI-1 蛋白和信使核糖核酸均下调（P <.05）。135 mg/dL 或更高浓度的葡萄糖诱导 sEng、sFlt-1 和白细胞介素 6 上调，而 VEGF 和 PlGF 下调。高血糖处理的 CTB 中 p38 磷酸化和 PPAR-γ上调（P <.05）。SB203580 或罗格列酮预处理可减轻葡萄糖诱导的 uPA 和 PAI-1 下调。
高血糖通过降低 uPA 和 PAI-1 表达、下调 VEGF 和 PlGF 以及上调 sEng、sFlt-1 和白细胞介素 6，破坏 CTB 的侵袭特征。SB203580 或罗格列酮预处理可减轻 CTB 功能障碍，提示应激信号的参与。
