title: Antihypertensive methyldopa, labetalol, hydralazine, and clonidine reversed tumour necrosis factor-α inhibited endothelial nitric oxide synthase expression in endothelial-trophoblast cellular networks.
abstract: Medications used to control hypertension in pregnancy also improve trophoblast and endothelial cellular interaction in vitro. Tumour necrosis factor-α (TNF-α) inhibits trophoblast and endothelial cellular interactions and simultaneously decreases endothelial nitric oxide synthase (eNOS) expression. This study investigated whether antihypertensive medications improved these cellular interactions by modulating eNOS and inducible nitric oxide synthase (iNOS) expression. Human uterine myometrial microvascular endothelial cells (UtMVECs) were pre-incubated with (or without) low dose TNF-α (0.5 ng/mL) or TNF-α plus soluble fms-like tyrosine kinase-1 (sFlt-1) (100 ng/mL). The endothelial cells were cultured on Matrigel. After endothelial cellular networks appeared, trophoblast derived HTR-8/SVneo cells were co-cultured in the presence of clinically relevant doses of methyldopa, labetalol, hydralazine or clonidine for 24 hours. Cells were retrieved from the Matrigel to extract mRNA and eNOS and iNOS expression were examined by quantitative PCR. Methyldopa, labetalol, hydralazine and clonidine reversed the inhibitory effect of TNF-α on eNOS mRNA expression. After pre-incubating endothelial cells with TNF-α and sFlt-1, all the medications except methyldopa lost their effect on eNOS mRNA expression. In the absence of TNF-α, antihypertensive medications did not change eNOS expression. The mRNA expression of iNOS was not affected by TNF-α or any medications. This study shows that selected antihypertensive medications used in the treatment of hypertension in pregnancy increase eNOS expression in vitro when induced by the inflammatory TNF-α. The anti-angiogenic molecule sFlt-1 may antagonise the potential benefit of these medications by interfering with the NOS pathway.
authors.affiliation: School of Medicine, Western Sydney University, Sydney, NSW, Australia.|Vascular Immunology Research Laboratory, The Heart Research Institute, University of Sydney, Sydney, NSW, Australia.
authors.forename: Bei
authors.identifier: 0000-0002-2101-3877
authors.initials: B
authors.lastname: Xu
authors.affiliation: School of Medicine, Western Sydney University, Sydney, NSW, Australia.
authors.forename: Gabriele
authors.initials: G
authors.lastname: Bobek
authors.affiliation: School of Medicine, Western Sydney University, Sydney, NSW, Australia.|Vascular Immunology Research Laboratory, The Heart Research Institute, University of Sydney, Sydney, NSW, Australia.|Renal Unit, Liverpool Hospital, Sydney, NSW, Australia.
authors.forename: Angela
authors.initials: A
authors.lastname: Makris
authors.affiliation: School of Medicine, Western Sydney University, Sydney, NSW, Australia.|Vascular Immunology Research Laboratory, The Heart Research Institute, University of Sydney, Sydney, NSW, Australia.
authors.forename: Annemarie
authors.initials: A
authors.lastname: Hennessy
mesh_terms: D000959:Antihypertensive Agents / Q000494:pharmacology*; D002478:Cells, Cultured; D003000:Clonidine / Q000494:pharmacology; D018920:Coculture Techniques; D017077:Culture Media, Conditioned; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects*; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects; D005260:Female; D006801:Humans; D006830:Hydralazine / Q000494:pharmacology; D000072179:Interleukin-1 Receptor-Like 1 Protein / Q000502:physiology; D007741:Labetalol / Q000494:pharmacology; D008750:Methyldopa / Q000494:pharmacology; D052250:Nitric Oxide Synthase Type III / Q000037:antagonists & inhibitors* / Q000235:genetics; D014327:Trophoblasts / Q000166:cytology / Q000187:drug effects*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D014599:Uterus / Q000166:cytology / Q000187:drug effects
chemical_list: D000959:Antihypertensive Agents; D017077:Culture Media, Conditioned; C079241:IL1RL1 protein, human; D000072179:Interleukin-1 Receptor-Like 1 Protein; D014409:Tumor Necrosis Factor-alpha; D006830:Hydralazine; D008750:Methyldopa; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; D003000:Clonidine; D007741:Labetalol
keywords: antihypertensive medication; clonidine; endothelial nitric oxide synthase; hydralazine; labetalol; methyldopa; preeclampsia; soluble fms-like tyrosine kinase-1; tumor necrosis factor-α