Vitamin D Reduces Oxidative Stress–Induced Procaspase-3/ROCK1 Activation and MP Release by Placental Trophoblasts

• Published in: The journal of clinical endocrinology and metabolism 2017-06, Vol.102 (6), p.2100-2110
• Main Authors: Xu, Jie, Jia, Xiuyue, Gu, Yang, Lewis, David F., Gu, Xin, Wang, Yuping
• Format: Article
• Language: English
• Subjects: Activation; Adolescent; Adult; Annexin V; Blotting, Western; Calcitriol; Calcitriol - pharmacology; Case-Control Studies; Caspase; Caspase 3 - drug effects; Caspase 3 - metabolism; Caspase-3; Caveolin; Caveolin 1 - drug effects; Caveolin 1 - metabolism; Caveolin-1; Cell-Derived Microparticles - drug effects; Cell-Derived Microparticles - secretion; Cleavage; Clinical s; Cobalt - pharmacology; Coiling; Female; Flow cytometry; Humans; Inflammation; Inflammatory response; Inhibition; Kinases; Nitric oxide; Nitric Oxide Synthase Type III - drug effects; Nitric Oxide Synthase Type III - metabolism; Nitric-oxide synthase; Oxidative stress; Oxidative Stress - drug effects; Placenta; Pre-Eclampsia; Pregnancy; Protein kinase; rho-Associated Kinases - drug effects; rho-Associated Kinases - metabolism; Shedding; Stress; Trophoblasts; Trophoblasts - drug effects; Trophoblasts - secretion; Vitamin D; Vitamins - pharmacology; Young Adult
• ISSN: 0021-972X; 1945-7197
• DOI: 10.1210/jc.2016-3753

Abstract Context: 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. Objective: To investigate if vitamin D could protect trophoblasts from oxidative stress–induced MP release. Design: Placental trophoblasts were isolated from uncomplicated and preeclamptic placentas. Effects of vitamin D on MP release induced by oxidative stress inducer CoCl2 were studied. Main Outcome Measures: 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. Results: 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. Conclusions: 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. Inhibition of caspase-3 cleavage and ROCK1 activation could be the potential protective mechanism(s) of 1,25(OH)2D3 to suppress microparticle shedding by placental trophoblasts.