Cawyer Chase, Afroze Syeda H, Drever Nathan, Allen Steven, Jones Richard, Zawieja David C, Kuehl Thomas, Uddin M Nasir
a Departments of Obstetrics & Gynecology , Texas A&M Health Science Center College of Medicine/Baylor Scott & White Health , Temple , Texas , USA.
b Medical Physiology , Texas A&M Health Science Center College of Medicine/Baylor Scott & White Health , Temple , Texas , USA.
Hypertens Pregnancy. 2016 May;35(2):159-69. doi: 10.3109/10641955.2015.1122035. Epub 2016 Mar 1.
Preeclampsia (preE) is a hypertensive disorder that occurs 20% in diabetic pregnancy. We have shown that hyperglycemia impairs cytotrophoblast cell (CTB) function. In this study, we assess apoptotic and anti-angiogenic signaling in excess glucose-induced CTBs.
Human extravillous CTBs (Sw. 71) were treated with 100, 150, 200, 300, or 400 mg/dL glucose for 48 h. Some cells were pretreated with a p38 inhibitor (SB203580) or a peroxisome proliferator-activated receptor gamma (PPARγ) ligand (rosiglitazone) or with D-mannitol. Cell lysates were utilized to measure p38 MAPK phosphorylation, PPARγ, Bcl-2-associated-X protein (Bax), anti-apoptotic Bcl-2, caspase-9, and cyclooxygenase-2 (Cox-2) expression by western blot. Levels of the vascular endothelial growth factor (VEGF), placental growth factor (PlGF), soluble fms-like tyrosine kinase-1 (sFlt-1), soluble endoglin (sEng), and interleukin 6 (IL-6) were measured in culture media using ELISA kits. Statistical comparisons were performed using analysis of variance with Duncan's post hoc test.
p38 phosphorylation and PPARγ were upregulated (p < 0.05) in CTBs treated with ≥150 mg/dL glucose compared to basal (100 mg/dL). Expressions of Bax/Bcl-2, Cox-2, and caspase-9 were upregulated (p < 0.05) in CTBs treated with ≥150 mg/dL glucose. Secretion of sFlt-1, sEng, and IL-6 was increased while VEGF and PIGF were decreased in CTB-treated ≥150 mg/dl of glucose (*p < 0.01 for each). SB203580 or rosiglitazone pretreatment significantly attenuated hyperglycemia-induced apoptotic and anti-angiogenic signaling. D-Mannitol had no effect.
Hyperglycemia induced apoptotic and anti-angiogenic signaling in CTBs. The observed diminution of hyperglycemia-induced signaling by SB203580 or rosiglitazone pretreatment suggests the involvement of apoptotic and anti-angiogenic signaling in CTB dysfunction.
子痫前期（preE）是一种高血压疾病，在糖尿病妊娠中发生率为20%。我们已表明高血糖会损害细胞滋养层细胞（CTB）功能。在本研究中，我们评估了高糖诱导的CTB中的凋亡和抗血管生成信号。
将人绒毛外CTB（Sw. 71）用100、150、200、300或400mg/dL葡萄糖处理48小时。一些细胞用p38抑制剂（SB203580）或过氧化物酶体增殖物激活受体γ（PPARγ）配体（罗格列酮）或D-甘露醇预处理。利用细胞裂解物通过蛋白质印迹法测量p38丝裂原活化蛋白激酶（MAPK）磷酸化、PPARγ、Bcl-2相关X蛋白（Bax）、抗凋亡蛋白Bcl-2、半胱天冬酶-9和环氧合酶-2（Cox-2）的表达。使用酶联免疫吸附测定（ELISA）试剂盒测量培养基中血管内皮生长因子（VEGF）、胎盘生长因子（PlGF）、可溶性fms样酪氨酸激酶-1（sFlt-1）、可溶性内皮糖蛋白（sEng）和白细胞介素6（IL-6）的水平。使用方差分析和邓肯事后检验进行统计比较。
与基础水平（100mg/dL）相比，用≥150mg/dL葡萄糖处理的CTB中p38磷酸化和PPARγ上调（p<0.05）。用≥150mg/dL葡萄糖处理的CTB中Bax/Bcl-2、Cox-2和半胱天冬酶-9的表达上调（p<0.05）。在处理葡萄糖≥150mg/dl的CTB中，sFlt-1、sEng和IL-6的分泌增加，而VEGF和PIGF减少（每项*p<0.01）。SB203580或罗格列酮预处理显著减弱了高血糖诱导的凋亡和抗血管生成信号。D-甘露醇无作用。
高血糖在CTB中诱导凋亡和抗血管生成信号。SB203580或罗格列酮预处理观察到的高血糖诱导信号的减弱表明凋亡和抗血管生成信号参与了CTB功能障碍。
