Xu Jie, Jia Xiuyue, Gu Yang, Lewis David F, Gu Xin, Wang Yuping
Department of Obstetrics and Gynecology, Louisiana State University Health Sciences Center, Shreveport, Louisiana 71130.
Department of Physiology, Harbin Medical University, Harbin, Heilongjiang 150086, China.
J Clin Endocrinol Metab. 2017 Jun 1;102(6):2100-2110. doi: 10.1210/jc.2016-3753.
Increased microparticle (MP) shedding by placental trophoblasts contributes to maternal vascular inflammatory response and endothelial dysfunction in preeclampsia. Vitamin D has beneficial effects in pregnancy; however, its effect on trophoblast MP release has not been investigated.
To investigate if vitamin D could protect trophoblasts from oxidative stress-induced MP release.
Placental trophoblasts were isolated from uncomplicated and preeclamptic placentas. Effects of vitamin D on MP release induced by oxidative stress inducer CoCl2 were studied.
Annexin V+ MPs were assessed by flow cytometry. Expression of caveolin-1, endothelial nitric oxide synthase (eNOS), procaspase-3, cleaved caspase-3, and Rho-associated coiled-coil protein kinase 1 (ROCK1) in trophoblasts and trophoblast-derived MPs were determined by Western blot.
Trophoblasts from preeclamptic pregnancies released significantly more MPs than cells from uncomplicated pregnancies (P < 0.01). CoCl2-induced increase in MP release was associated with upregulation of caveolin-1 and downregulation of eNOS expression in trophoblasts (P < 0.05), which could be attenuated by 1,25(OH)2D3. Moreover, 1,25(OH)2D3 could also inhibit CoCl2-induced procaspase-3 cleavage and ROCK1 activation in trophoblasts. Consistently, CoCl2-induced upregulation of procaspase-3, cleaved caspase-3, and ROCK1 expression in trophoblast-derived MPs were also reduced in cells treated with 1,25(OH)2D3.
Placental trophoblasts from preeclamptic pregnancies released more MP than cells from uncomplicated pregnancies. Oxidative stress-induced increase in MP shedding is associated with upregulation of caveolin-1 and downregulation of eNOS expression in placental trophoblasts. Inhibition of caspase-3 cleavage and ROCK1 activation, together with upregulation of eNOS expression, could be the potential cellular/molecular mechanism(s) of vitamin D protective effects on placental trophoblasts.
胎盘滋养层细胞释放的微粒（MP）增加会导致子痫前期孕妇的血管炎症反应和内皮功能障碍。维生素D在孕期具有有益作用；然而，其对滋养层细胞释放MP的影响尚未得到研究。
研究维生素D是否能保护滋养层细胞免受氧化应激诱导的MP释放。
从正常和子痫前期胎盘分离胎盘滋养层细胞。研究维生素D对氧化应激诱导剂氯化钴（CoCl2）诱导的MP释放的影响。
通过流式细胞术评估膜联蛋白V阳性MP。通过蛋白质免疫印迹法测定滋养层细胞和滋养层来源的MP中小窝蛋白-1、内皮型一氧化氮合酶（eNOS）、半胱天冬酶原-3、裂解的半胱天冬酶-3和Rho相关卷曲螺旋蛋白激酶1（ROCK1）的表达。
子痫前期妊娠的滋养层细胞释放的MP明显多于正常妊娠的细胞（P < 0.01）。CoCl2诱导的MP释放增加与滋养层细胞中小窝蛋白-1的上调和eNOS表达的下调有关（P < 0.05），而1,25(OH)2D3可使其减弱。此外，1,25(OH)2D3还可抑制CoCl2诱导的滋养层细胞中半胱天冬酶原-3的裂解和ROCK1的激活。同样，在用1,25(OH)2D3处理的细胞中，CoCl2诱导的滋养层来源的MP中半胱天冬酶原-3、裂解的半胱天冬酶-3和ROCK1表达的上调也有所减少。
子痫前期妊娠的胎盘滋养层细胞比正常妊娠的细胞释放更多的MP。氧化应激诱导的MP释放增加与胎盘滋养层细胞中小窝蛋白-1的上调和eNOS表达的下调有关。抑制半胱天冬酶-3的裂解和ROCK1的激活，以及eNOS表达的上调，可能是维生素D对胎盘滋养层细胞保护作用的潜在细胞/分子机制。
