Sildenafil Citrate Downregulates PDE5A mRNA Expression in Women with Recurrent Pregnancy Loss without Altering Angiogenic Factors—A Preliminary Study

In our previous study, we showed that sildenafil citrate (SC), a selective PDE5A blocker, modulated NK cell activity in patients with recurrent pregnancy loss, which correlated with positive pregnancy outcomes. It was found that NK cells had a pivotal role in decidualization, angiogenesis, spiral ar...

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Autores principales:	Monika Kniotek, Aleksander Roszczyk, Michał Zych, Małgorzata Wrzosek, Monika Szafarowska, Radosław Zagożdżon, Małgorzata Jerzak
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	RPL NK cells sildenafil PDE5A VEGF-A angiotensin Medicine R
Acceso en línea:	https://doaj.org/article/8b1d03903ad149e29fd0539aedbc7c31
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spelling	oai:doaj.org-article:8b1d03903ad149e29fd0539aedbc7c312021-11-11T17:42:34ZSildenafil Citrate Downregulates PDE5A mRNA Expression in Women with Recurrent Pregnancy Loss without Altering Angiogenic Factors—A Preliminary Study10.3390/jcm102150862077-0383https://doaj.org/article/8b1d03903ad149e29fd0539aedbc7c312021-10-01T00:00:00Zhttps://www.mdpi.com/2077-0383/10/21/5086https://doaj.org/toc/2077-0383In our previous study, we showed that sildenafil citrate (SC), a selective PDE5A blocker, modulated NK cell activity in patients with recurrent pregnancy loss, which correlated with positive pregnancy outcomes. It was found that NK cells had a pivotal role in decidualization, angiogenesis, spiral artery remodeling, and the regulation of trophoblast invasion. Thus, in the current study, we determined the effects of SC on angiogenic factor expression and production, as well as idNK cell activity in the presence of nitric synthase blocker L-NMMA. Methods: NK cells (CD56<sup>+</sup>) were isolated from the peripheral blood of 15 patients and 15 fertile women on MACS columns and cultured in transformation media containing IL-15, TGF-β, and AZA—a methylation agent—for 7 days in hypoxia (94% N<sub>2</sub>, 1% O<sub>2</sub>, 5% CO<sub>2</sub>). Cultures were set up in four variants: (1) with SC, (2) without SC, (3) with NO, a synthase blocker, and (4) with SC and NO synthase blocker. NK cell activity was determined after 7 days of culturing as CD107a expression after an additional 4h of stimulation with K562 erythroleukemia cells. The expression of the <i>PDE5A, VEGF-A, PIGF, IL-8,</i> and <i>RENBP</i> genes was determined with quantitative real-time PCR (qRT-PCR) using TaqMan probes and ELISA was used to measure the concentrations of VEGF-A, PLGF, IL-8, Ang-I, Ang-II, IFN–γ proteins in culture supernatants after SC supplementation. Results: SC downregulated <i>PDE5A</i> expression and had no effect on other studied angiogenic factors. <i>VEGF-A</i> expression was increased in RPL patients compared with fertile women. Similarly, VEGF production was enhanced in RPL patients’ supernatants and SC increased the concentration of PIGF in culture supernatants. SC did not affect the expression or concentration of other studied factors, nor idNK cell activity, regardless of NO synthase blockade.Monika KniotekAleksander RoszczykMichał ZychMałgorzata WrzosekMonika SzafarowskaRadosław ZagożdżonMałgorzata JerzakMDPI AGarticleRPLNK cellssildenafilPDE5AVEGF-AangiotensinMedicineRENJournal of Clinical Medicine, Vol 10, Iss 5086, p 5086 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	RPL NK cells sildenafil PDE5A VEGF-A angiotensin Medicine R
spellingShingle	RPL NK cells sildenafil PDE5A VEGF-A angiotensin Medicine R Monika Kniotek Aleksander Roszczyk Michał Zych Małgorzata Wrzosek Monika Szafarowska Radosław Zagożdżon Małgorzata Jerzak Sildenafil Citrate Downregulates PDE5A mRNA Expression in Women with Recurrent Pregnancy Loss without Altering Angiogenic Factors—A Preliminary Study
description	In our previous study, we showed that sildenafil citrate (SC), a selective PDE5A blocker, modulated NK cell activity in patients with recurrent pregnancy loss, which correlated with positive pregnancy outcomes. It was found that NK cells had a pivotal role in decidualization, angiogenesis, spiral artery remodeling, and the regulation of trophoblast invasion. Thus, in the current study, we determined the effects of SC on angiogenic factor expression and production, as well as idNK cell activity in the presence of nitric synthase blocker L-NMMA. Methods: NK cells (CD56<sup>+</sup>) were isolated from the peripheral blood of 15 patients and 15 fertile women on MACS columns and cultured in transformation media containing IL-15, TGF-β, and AZA—a methylation agent—for 7 days in hypoxia (94% N<sub>2</sub>, 1% O<sub>2</sub>, 5% CO<sub>2</sub>). Cultures were set up in four variants: (1) with SC, (2) without SC, (3) with NO, a synthase blocker, and (4) with SC and NO synthase blocker. NK cell activity was determined after 7 days of culturing as CD107a expression after an additional 4h of stimulation with K562 erythroleukemia cells. The expression of the <i>PDE5A, VEGF-A, PIGF, IL-8,</i> and <i>RENBP</i> genes was determined with quantitative real-time PCR (qRT-PCR) using TaqMan probes and ELISA was used to measure the concentrations of VEGF-A, PLGF, IL-8, Ang-I, Ang-II, IFN–γ proteins in culture supernatants after SC supplementation. Results: SC downregulated <i>PDE5A</i> expression and had no effect on other studied angiogenic factors. <i>VEGF-A</i> expression was increased in RPL patients compared with fertile women. Similarly, VEGF production was enhanced in RPL patients’ supernatants and SC increased the concentration of PIGF in culture supernatants. SC did not affect the expression or concentration of other studied factors, nor idNK cell activity, regardless of NO synthase blockade.
format	article
author	Monika Kniotek Aleksander Roszczyk Michał Zych Małgorzata Wrzosek Monika Szafarowska Radosław Zagożdżon Małgorzata Jerzak
author_facet	Monika Kniotek Aleksander Roszczyk Michał Zych Małgorzata Wrzosek Monika Szafarowska Radosław Zagożdżon Małgorzata Jerzak
author_sort	Monika Kniotek
title	Sildenafil Citrate Downregulates PDE5A mRNA Expression in Women with Recurrent Pregnancy Loss without Altering Angiogenic Factors—A Preliminary Study
title_short	Sildenafil Citrate Downregulates PDE5A mRNA Expression in Women with Recurrent Pregnancy Loss without Altering Angiogenic Factors—A Preliminary Study
title_full	Sildenafil Citrate Downregulates PDE5A mRNA Expression in Women with Recurrent Pregnancy Loss without Altering Angiogenic Factors—A Preliminary Study
title_fullStr	Sildenafil Citrate Downregulates PDE5A mRNA Expression in Women with Recurrent Pregnancy Loss without Altering Angiogenic Factors—A Preliminary Study
title_full_unstemmed	Sildenafil Citrate Downregulates PDE5A mRNA Expression in Women with Recurrent Pregnancy Loss without Altering Angiogenic Factors—A Preliminary Study
title_sort	sildenafil citrate downregulates pde5a mrna expression in women with recurrent pregnancy loss without altering angiogenic factors—a preliminary study
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/8b1d03903ad149e29fd0539aedbc7c31
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Sildenafil Citrate Downregulates PDE5A mRNA Expression in Women with Recurrent Pregnancy Loss without Altering Angiogenic Factors—A Preliminary Study

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