Resolvin D1 reverses reactivity and Ca²⁺ sensitivity induced by ET-1, TNF-α, and IL-6 in the human pulmonary artery

Abstract

monary hypertension (PH) is a rare and progressive disease characterized by an inflammatory status and vessel wall remodeling, resulting in increased pulmonary artery resistance. During the last decade, treatments have been proposed; most of them target the endothelial pathways that stimulate smooth muscle cell relaxation. However, PH remains associated with significant morbidity. We hypothesized that inflammation plays a crucial role in the severity of the abnormal vasoconstriction in PH. The goal of this study was to assess the effects of resolvin D1 (RvD1), a potent anti-inflammatory agent, on the pharmacological reactivity of human pulmonary arteries (HPAs) via an in vitro model of induced hyperreactivity. The effects of RvD1 and monoacylglyceride compounds were measured on contractile activity and Ca 2ϩ sensitivity developed by HPAs that had been pretreated (or not) under proinflammatory conditions with either 10 ng/ml TNF-␣ or 10 ng/ml IL-6 or under hyperreactive conditions with 5 nM endothelin-1. The results demonstrated that, compared with controls, 24-h pretreatment with TNF-␣, IL-6, or endothelin-1 increased reactivity and Ca 2ϩ sensitivity of HPAs as revealed by agonist challenges with 80 mM KCl, 1 M serotonin (5-hydroxytryptamine), 30 nM U-46619, and 1 M phorbol 12,13-dibutyrate. However, 300 nM RvD1 as well as 1 M monoacylglyceride-docosapentaenoic acid monoglyceride strongly reversed the overresponsiveness induced by both proinflammatory and hyperreactive treatments. In pretreated pulmonary artery smooth muscle cells, Western blot analyses revealed that RvD1 treatment decreased the phosphorylation level of CPI-17 and expression of transmembrane protein member 16A while increasing the detection of G protein-coupled receptor 32. The present data demonstrate that RvD1, a trihydroxylated docosahexaenoic acid derivative, decreases induced overreactivity in HPAs via a reduction in CPI-17 phosphorylation and transmembrane protein member 16A expression. pulmonary hypertension; docosapentanenoic acid monoacylglyceride; endothelin; resolvin D1; CPI-17; endothelin-1; tumor necrosis factor-␣; interleukin-6

MAG-DHA, Precursor of D-Series Resolvins, Induces Powerful Resolution of Various Components of Pulmonary Hypertension Induced By Monocrotaline in Rats

“…The DHA metabolite Resolvin D1 has recently been shown to reduce inflammation markers and hyperreactivity in human pulmonary artery upon in-vitro treatment [17]. To strengthen these reports, the current (A) Experimental design and treatments.…”

“…In a previous study, we also demonstrated that MAG-DHA needs to be metabolized to exert its beneficial action, and that one of its metabolites, resolving D1 (RvD1), leads an enhanced expression of GPR32 [17], previously identified as the RvD1 receptor [24]. In order to further assess the manner in which MAG-DHA may counteract the effects of MCT treatment on the expression of these key biological markers, PASM cell lysates were fractionated using differential ultracentrifugations from which microsomal fractions were used to detect expression of membrane proteins (TMEM16A and GPR32) while cytosolic fractions were used to detect the expression of HIF1-α.…”

“…MAG-DHA is neutral lipid which is very well absorbed by tissues [29] and becomes a DHA donor upon the action of MAG-lipase and the precursor of several omega-3 derivatives such as resolvins and protectins upon the action of 15-LOX and 5-LOX [10]. Resolvins have been reported to reduce pro-inflammatory markers [17,30], namely TNFα and IL-1β, and to prevent systemic hypertension in rats [31]. Furthermore, RvD1, A trihydroxylated DHA derivative has recently been reported to reverse the pharmacological reactivity and Ca 2+ sensitivity induced by ET-1 and IL-6 in human pulmonary artery [17].…”

“…Resolvins have been reported to reduce pro-inflammatory markers [17,30], namely TNFα and IL-1β, and to prevent systemic hypertension in rats [31]. Furthermore, RvD1, A trihydroxylated DHA derivative has recently been reported to reverse the pharmacological reactivity and Ca 2+ sensitivity induced by ET-1 and IL-6 in human pulmonary artery [17].…”

“…Our group has already investigated the impact of different monoacylglyceride compounds and shown that monoacylglyceride docosahexaenoic acid (MAG-DHA) [15], monoacylglyceride docosapentaenoic acid (MAG-DPA) [16], and D-series resolvins such as Resolvin D1 were able to abolish the pro-inflammatory effects of TNF-α and IL-6 as well as the induced hyperreactivity and calcium sensitivity in human pulmonary arteries [17].…”