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. 2020 Aug 29;16(3):453–461. doi: 10.1007/s11302-020-09721-2

Impaired UTP-induced relaxation in the carotid arteries of spontaneously hypertensive rats

Takayuki Matsumoto 1,✉, Mihoka Kojima 1, Keisuke Takayanagi 1, Tomoki Katome 1, Kumiko Taguchi 1, Tsuneo Kobayashi 1,✉
PMCID: PMC7524952  PMID: 32862324

Abstract

Uridine 5′-triphosphate (UTP) has an important role as an extracellular signaling molecule that regulates inflammation, angiogenesis, and vascular tone. While chronic hypertension has been shown to promote alterations in arterial vascular tone regulation, carotid artery responses to UTP under hypertensive conditions have remained unclear. The present study investigated carotid artery responses to UTP in spontaneously hypertensive rats (SHR) and control Wistar Kyoto rats (WKY). Accordingly, our results found that although UTP promotes concentration-dependent relaxation in isolated carotid artery segments from both SHR and WKY after pretreatment with phenylephrine, SHR exhibited significantly lower arterial relaxation responses compared with WKY. Moreover, UTP-induced relaxation was substantially reduced by endothelial denudation and by the nitric oxide (NO) synthase inhibitor NG-nitro-L-arginine in both SHR and WKY. The difference in UTP-induced relaxation between both groups was abolished by the selective P2Y2 receptor antagonist AR-C118925XX and the cyclooxygenase (COX) inhibitor indomethacin but not by the thromboxane-prostanoid receptor antagonist SQ29548. Furthermore, we detected the release of PGE2, PGF2α, and PGI2 in the carotid arteries of SHR and WKY, both at baseline and in response to UTP. UTP administration also increased TXA2 levels in WKY but not SHR. Overall, our results suggest that UTP-induced relaxation in carotid arteries is impaired in SHR perhaps due to impaired P2Y2 receptor signaling, reductions in endothelial NO, and increases in the levels of COX-derived vasoconstrictor prostanoids.

Electronic supplementary material

The online version of this article (10.1007/s11302-020-09721-2) contains supplementary material, which is available to authorized users.

Keywords: Carotid artery, Hypertension, P2Y2, Relaxation, UTP

Introduction

Adenosine triphosphate (ATP), adenosine diphosphate (ADP), uridine triphosphate (UTP), uridine diphosphate (UDP), and uridine adenosine tetraphosphate (Up4A) are among the nucleotides released from cellular sources in response to chemical or mechanical stress, platelet activation, and cell injury and/or death [1–3]. Accordingly, extracellular nucleotides function as signaling molecules and play pivotal roles in cardiovascular biology and diseases [2–4]. Extracellular purine nucleotides have been found to induce signaling via purinergic receptors (purinoceptors), including both ion channels (P2X receptors) and G protein-coupled receptors (GPCRs; P2Y receptors) [2, 3, 5, 6]. Nonetheless, the physiologic roles of extracellular nucleotides have yet to be comprehensively understood considering the multiple conflicting reports documenting both harmful and protective effects. Cell type–specific expression of specific P2Y receptors determines cellular responses to nucleotides, with P2Y2 and P2Y4 being the major receptors for UTP [2, 3, 5, 6]. In the cardiovascular system, P2Y receptor activation by UTP and related compounds increases inotropy, controls hypertrophic growth of cardiomyocytes, and modulates the response to pressure overload in the heart [7–10]; facilitates proliferation, migration, and calcification of vascular smooth muscle cells [2, 3, 11, 12]; and promotes the release of various factors, such as endothelium-derived relaxing factors (EDRFs) (e.g., nitric oxide (NO), PGI2, and endothelium-derived hyperpolarizing factors (EDHFs)) and endothelium-derived contracting factors (EDCFs) (e.g., vasoconstrictor prostanoids and Up4A) [13–18]. Extracellular UTP exerts not only vasoconstrictive but also vasorelaxant effects [2, 3, 15–20]. These discordant findings may be related to the nature of the target species, vessel types, and specific study conditions, including duration of disease, age, and sex.

Hypertension has been associated with complex functional alterations in endothelial cells, including decreased EDRF and increased EDCF production, as well as altered responses to various vasoactive factors generated by vascular smooth muscle cells [1, 21–24]. Taken together, these dysfunctional responses may contribute to elevated peripheral vascular resistance and/or the development of systematic complications in renal, coronary, or cerebral circulation. The carotid arteries are major conduits that supply blood to the brain. Although several reports have suggested that hypertension may promote dysfunctional responses in the carotid arteries [25–29], very few studies have investigated the responses of these arteries to extracellular nucleotides [27].

Only a handful of studies have focused on the role of UTP and its impact on hypertension within the carotid arteries. The present study therefore aimed to investigate whether spontaneously hypertensive rats (SHR) experience alterations in UTP-mediated carotid artery response by comparing them with age-matched control Wistar Kyoto rats (WKY), as well as determine whether this response was associated with NO and cyclooxygenase (COX)-derived mediators.

Materials and methods

Animals and experimental design

Four-week-old male rats (SHR and WKY) were supplied by Hoshino Laboratory Animals, Inc. (Ibaraki, Japan). All experimental procedures were approved by the Hoshi University Animal Care and Use Committee, and all studies were conducted in accordance with the Guide for the Care and Use of Laboratory Animals, published by the US National Institutes of Health, and the Guide for the Care and Use of Laboratory Animals adopted by the Committee on the Care and Use of Laboratory Animals of Hoshi University (accredited by the Ministry of Education, Culture, Sports, Science and Technology, Japan). At 20–22 weeks of age, SHR (n = 17) had higher systolic blood pressure than WKY (n = 18) (184 ± 3 mmHg vs.115 ± 3 mmHg, respectively; p < 0.0001) measured using a blood pressure analyzer (BP-98A; Softron, Tokyo, Japan) via the tail-cuff method as previously described [30–34]. No difference in body weight was noted between SHR and WKY at the time of sacrifice (392.9 ± 6.1 g vs. 407.1 ± 4.0 g, respectively; p > 0.05). In all experiments, non-fasted rats were anesthetized with isoflurane via a nose cone for surgical procedures and subsequently euthanized through thoracotomy and exsanguination. After euthanasia, the common carotid arteries were isolated and placed in oxygenated, ice-cold modified Krebs–Henseleit solution (KHS) (118.0 mM NaCl, 4.7 mM KCl, 25.0 mM NaHCO3, 1.8 mM CaCl2, 1.2 mM NaH2PO4, 1.2 mM MgSO4, and 11.0 mM glucose). Subsequently, the artery was carefully cleaned and cut into rings to facilitate investigation of vascular functional (i.e., isometric force) and molecular responses (i.e., prostanoid release).

Measurement of isometric force

Isometric force was measured as previously described [32, 33]. Briefly, after cleaning, 2-mm-long carotid artery ring segments of the carotid arteries were cut and suspended using a pair of stainless steel pins in a well-oxygenated (95% O2, 5% CO2) organ bath containing KHS at 37 °C. Vascular isometric force was monitored using an isometric transducer (TB-611 T; Nihon Kohden, Tokyo, Japan) linked to a PowerLab recording system (AD Instruments, Australia). Arterial integrity was confirmed through experiments involving contraction, carried out at high levels of K+ (80 mM) followed by phenylephrine (PE) (10−6 M), and relaxation with acetylcholine (ACh) (10−6 M for arteries with intact endothelium or 10−5 M for endothelial-denuded arteries). After washing and re-stabilization, concentration–response curves for UTP (10−8–10−4 M) or UDP (10−8–10−4 M) were generated from arteries precontracted with 10−6 M PE. To rule out desensitization via purinoceptor stimulation when comparing SHR and WKY, we investigated the first concentration−response curve for UTP/other ligands in each arterial ring. As displayed in supplemental Fig. S1, high K+, PE, or ACh were successfully washed out and returned to basal tension. Moreover, successful PE-induced contraction was observed in the artery. To explore UTP-mediated signal transduction, UTP-mediated relaxation was evaluated under several experimental conditions, including endothelial denudation. Denudation was accomplished through a 1-min infusion of a solution of 3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate solution (CHAPS, 0.1%) [14, 32, 33]. We also examined P2Y2 receptor antagonism by AR-C118925XX (10−5 M), inhibition of NO synthase (NOS) with NG-nitro-L-arginine (L-NNA; 10−4 M), inhibition of COX by indomethacin (10−5 M), and antagonism of thromboxane-prostanoid (TP) receptor by SQ29548 (3 × 10−6 M). Each inhibitor/antagonist was applied 30 min prior to PE application and was present throughout the experiment. Absolute force of 10−6 M PE-induced contraction (mN/mg) did not significantly differ between SHR and WKY groups in each condition (Supplemental Table S1).

Measurement of prostanoid release

Prostanoid release was measured essentially as described in our previous reports [30, 31, 35]. Briefly, carotid arterial rings (6-mm-long segments) were incubated in 1.0 mL KHS at 37 °C. The rings were then rapidly transferred to siliconized tubes containing 0.5 mL KHS in the absence (control) or presence of UTP (10−5 M) at 37 °C for 10 min. After the rings were removed and weighed, the tubes containing released mediators were flash-frozen in liquid nitrogen and stored at − 80 °C for later analysis. Prostanoids or their stable metabolites, including PGE2, PGF2α, thromboxane B2, (a stable metabolite of TXA2), and 6-keto PGF1α (a stable metabolite of PGI2), were measured using a commercially available enzyme immunoassay kits (Cayman Chemical, Ann Arbor, MI, USA) as described in the manufacturer’s instructions. Prostanoid release was presented as pg/mg or ng/mg wet weight of carotid arterial ring tissue.

Data analysis and statistics

Data are expressed as means ± SE, with n representing the number of animals used in the experiments. Each relaxation response is presented as a percentage of the PE-induced contraction. Statistical comparisons between the two groups were performed using Student’s t test. One-way analysis of variance (ANOVA) followed by Tukey’s testing was used for comparisons among three or more groups. Concentration–response curves were statistically evaluated using two-way repeated measures ANOVA followed by Bonferroni post hoc testing. A p value of < 0.05 indicated statistical significance.

Results and discussion

Reports have shown that functional phenotypes induced by extracellular nucleotides and/or dinucleotides differed according to vessel type, species, and several diseases, including hypertension [2–4, 6]. For instance, UTP promoted endothelium-dependent arterial relaxation in mouse aorta, rat mesenteric arterial bed, and mouse coronary arteries [16, 18, 36, 37]. We had previously reported that SHR exhibited weaker ATP- and ADP-induced relaxation of the superior mesenteric arteries compared with age-matched control WKY, whereas both groups had similar adenosine-induced relaxation [38]. Moreover, studies have shown that rats with deoxycorticosterone salt-induced hypertension exhibited region-specific alteration in Up4A-induced arterial contractions, which were increased in basilar, renal, and femoral arteries, reduced in small mesenteric arteries, and unchanged in thoracic aorta and pulmonary arteries (vs. control rats) [39, 40]. Furthermore, P2Y6 receptor was the predominant contractile receptor for UTP- and UDP-induced contraction in mouse coronary arteries, whereas P2Y2 receptors mediated endothelium-dependent relaxation in larger diameter segments of the left descending coronary artery [36]. Another study showed that UTP induced endothelium-dependent contraction in porcine pancreatic arteries perhaps through the P2Y2 and P2Y4 receptors [41]. Thus, several pieces of functional evidence are required to comprehensively understand the role of extracellular nucleotide in vascular tone regulation during hypertension. As shown in Fig. 1a, UTP promoted carotid artery relaxation in both SHR and WKY, although SHR showed significantly lesser relaxation responses to increasing UTP concentrations in the carotid arteries compared with WKY. Moreover, no relaxation responses were observed in endothelial-denuded carotid arteries from both SHR and WKY (Fig. 1b). These results suggest that UTP-induced rat carotid arterial relaxation was endothelium-dependent and that UTP-mediated signaling in the endothelium was impaired in SHR arteries.

Fig. 1.

Fig. 1

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Uridine 5′-triphosphate (UTP) induces carotid arteries relaxation in spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY): impact of endothelial denudation, NOS inhibitor, and COX inhibitor, thromboxane-prostanoid receptor antagonist, and P2Y2 receptor antagonist, applied 30 min prior to PE. a Concentration−response curve showing relaxation (%PE) according to UTP concentration in the carotid arteries from WKY (open symbols) and SHR (closed symbols). b–f Impact of endothelial denudation (EC-), L-NNA (10−4 M), indomethacin (10−5 M), SQ29548 (3 × 10−6 M), and AR-C118925XX (10−5 M). g Concentration−response curve showing relaxation (%PE) according to uridine 5′-diphosphate (UDP) concentration in the carotid arteries from WKY (open symbols) and SHR (closed symbols). Each point represents the mean ± SEM of the maximal response to each concentration, n = 5–9. *p < 0.05, vs. WKY, using two-way repeated measures ANOVA followed by Bonferroni’s post hoc test

Several endothelium-derived vasoactive substances have been identified, with their complex signaling affecting vascular function [1, 21]. NO, PGI2, and EDHF have been known as EDRFs [21]. Accordingly, several reports have suggested that UTP-induced relaxation was mediated by not only NO but also EDHF in different arteries [18, 42, 43]. The results of the present study showed that under L-NNA-induced NOS inhibition, UTP induced very limited relaxation in carotid arteries of WKY. In contrast, limited contraction but not relaxation had been observed in SHR (Fig. 1c). Given that UTP-induced relaxation was largely blocked by L-NNA in the rat carotid artery, NO rather than EDHF can be considered the main relaxing factor when UTP was used to stimulate the endothelium.

COX-derived prostanoids play an important role in vascular tone regulation during (patho)physiological states, including hypertension [1, 44, 45]. Although PGI2 functions as another EDRF through the IP receptor [21], it may function as an EDCF through the activation of the TP receptor in some situations, including hypertension [21, 22]. To investigate whether COX-derived prostanoids could contribute to alterations in UTP-induced relaxation in SHR, we investigated the concentration–response curve for UTP in the presence of indomethacin, a nonselective COX inhibitor. Accordingly, our results showed that indomethacin treatment promoted an increase in UTP-induced relaxation in SHR carotid arteries but had no modulating effects on UTP-induced relaxation in WKY (Figs. 1a vs. 1d). Indomethacin treatment abolished the difference in UTP-induced relaxation between SHR and WKY (Fig. 1d). Studies have shown that increased activities of both COX-1 and COX-2 play a role in vascular dysfunction [1, 21]. After comparing SHR and WKY at 37–41 weeks old, the present study found differences in UTP-induced relaxation of the intact carotid arteries (Supplemental Fig. S2a) and in the presence of selective COX-1 inhibitor valeroyl salicylate (Supplemental Fig. S2b). However, this was abolished by the selective COX-2 inhibitor NS398 (Supplemental Fig. S2c), suggesting that the difference in UTP-induced relaxation may be determined by COX-2 rather than COX-1 activities in the carotid arteries. Moreover, SHR had higher levels of PGE2 (Fig. 2a), PGF2α (Fig. 2b), and PGI2 metabolite, 6-keto PGF1α (Fig. 2c) in the carotid arteries compared with WKY both at baseline and in response to UTP. Furthermore, UTP increased the levels of TXA2 metabolite TXB2 in WKY, although WKY and SHR had similar levels at baseline and in the presence of UTP (Fig. 2d). An earlier study reported that 15- to 16-week-old SHR had higher levels of ACh-induced TXB2, 6-keto PGF1α, and PGF2α in the aortas than age-matched WKY [46]. We recently found that 15- to 18-week-old SHR exhibited greater release of PGE2, PGF2α, and 6-keto PGF1α in the femoral arteries than age-matched WKY in response to noradrenaline [34]. Likewise, Gluais et al. [44] reported that ACh-induced endothelium-dependent release of TXA2, PGF2α, PGE2, PGI2, and most likely PGH2 in aortas isolated from 1-year-old WKY and SHR and that SHR had significantly higher PGI2 release in the aortas compared to WKY. They also reported that the calcium ionophore A23187 could induce prostanoid release in aortas isolated from 1-year-old WKY and SHR and that A23187 promoted a significantly higher release of PGI2 and TXA2 in SHR aortas compared to ACh [45]. The differences in prostanoid release observed after comparing the responses in two different arteries from SHR may be secondary to the experimental conditions (e.g., stimulants and exposed time), region, and/or age differences. Evaluation of prostanoid release in WKY showed that only TXB2 levels significantly increased in response to UTP stimulation (Fig. 2d). Given that UTP-induced relaxation had not been altered by indomethacin in WKY carotid arteries, we concluded that TXA2 may not contribute to UTP-induced relaxation even with increased levels. Likewise, our findings suggested that PGI2 may not play a significant role in the UTP-induced relaxation of WKY carotid arteries. Indeed, this is supported by data showing that the stable PGI2 analog beraprost-induced contraction but not relaxation in carotid arteries obtained from WKY and SHR at 40 or 41 weeks old, with both groups exhibiting similar contraction (Supplemental Fig. S3a). By contrast, the levels of PGE2, PGF2α, and PGI2 metabolites were increased in SHR carotid arteries, while UTP-induced relaxation was increased in response to indomethacin, with both groups showing similar responses under COX inhibition. In fact, PGE2 (Supplemental Fig. S3b), thromboxane analog U46619 (Supplemental Fig. S3c), and PGF2α (Supplemental Fig. S3d) induced carotid artery contraction in SHR and WKY. Although both SHR and WKY exhibited similar PGE2- and U46619-induced contractions, SHR showed greater PGF2α-induced contraction than WKY (Supplemental Fig. S3). Furthermore, the difference in UTP-induced contraction had not been abolished by the TP receptor antagonist SQ29548 (Fig. 1e). Taken together, the aforementioned data and relevant evidences suggest that UTP-induced relaxation was due to the NO component rather than EDHF and that the vasoconstrictor prostanoids in SHR arteries are unmasked and subsequently weaken the NO-mediated relaxant component, resulting in impaired UTP-induced relaxation. This notion is supported by substantial evidence suggesting that constrictor prostanoids could interrupt endothelium-dependent relaxation [30, 31] and that COX inhibitors could improve endothelium-dependent relaxation in SHR arteries [47]. Notably, the present study has some limitations. Although the release of prostanoids may interact with PE and potentially augment contraction, any effect of PE itself to release constrictor prostanoids is currently unknown because our data on prostanoid release are not presented for PE. Further investigations will be required to determine whether a positive interaction between PE and UTP is present upon the release of prostanoids. Although the exact mechanism by which prostanoids modulate UTP-induced relaxation in SHR carotid arteries remains unclear, EP and/or FP receptors rather than TP receptors have been suggested to play a role. Further investigations on the relationship between UTP-induced relaxation and prostanoid receptors will thus be required.

Fig. 2.

Fig. 2

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Release of prostanoids in carotid arteries from spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY). The levels of PGE2 (a), PGF2α (b), 6-keto PGF1α (c), and (d) TXB2 in carotid arteries from SHR and WKY at baseline and in response to uridine 5′-triphosphate (10−5 M). Data are presented as means ± SEM; n = 6. *p < 0.05, #p < 0.05, for WKY vs. SHR

UTP, a ligand for both P2Y2 and P2Y4 receptors [2–6], may indirectly activate P2Y6 receptors given that UTP is hydrolyzed to UDP, a ligand for the P2Y6 receptor [2–6]. We had recently observed that UDP-induced contraction rather than relaxation in rat carotid artery [35]. As shown in Fig. 1f, the selective P2Y2 antagonist AR-C118925XX abolished the difference in UTP-induced relaxation between SHR and WKY. Furthermore, UDP elicited slight relaxation at intermediate concentrations (i.e., 3 × 10−7 to 10−5 M) and contraction at higher concentrations in the carotid arteries of SHR and WKY (Fig. 1g). Accordingly, UDP-induced responses were similar in both groups (Fig. 1g). Although the contribution of the UTP-induced P2Y4 receptor remains unclear due to the lack of specific commercially available antagonist, P2Y4 receptors have been associated with contractile effects in rat arteries [48, 49]. While alterations in P2Y2 receptor signaling following UTP stimulation may be partly attributed to impaired UTP-induced relaxation responses in SHR carotid arteries, the contribution of other receptors, including P2Y4, P2Y6 (activated by UDP hydrolyzed from UTP in the vessel), and AR-C118925XX-resistant unknown receptors, could not be ruled out in the present study. Indomethacin-induced blockage of prostanoid synthesis resulted in similar UTP-induced relaxation in both two groups, suggesting that SHR and WKY had similar UTP-induced relaxant components. However, we are still unable to determine which receptor (P2Y2 and/or AR-C118925XX-resistant receptors) contributed to such a relaxant component, while complex interactions between relaxant (i.e., NO and EDHF) and contractile (i.e., COX-derived vasocontractile prostanoids) components may be present in SHR carotid arteries. Further investigations on the exact relationship between carotid arterial relaxation, receptor stimulation, and UTP kinetics in hypertensive vasculature will undoubtedly be required.

Conclusions

The present study reported that SHR had lower UTP-induced relaxation in the carotid arteries compared with WKY, which could perhaps be related to P2Y2 receptor-mediated signaling, NO signaling, and production and release of vasoconstrictor prostanoids. Although the precise pathophysiological significance of UTP signaling with respect to spontaneous hypertension has yet to be fully clarified, our findings suggest that the UTP-mediated vasomotor response could be a viable target for therapies developed to treat hypertension-associated vascular complications.

Electronic supplementary material

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Acknowledgments

We would like to thank Marina Ito, Yurika Ezaki, Amane Kurakata, Yuzuki Sato, Tamayo Hashimoto, Yurina Mae, and Hiyori Yokoyama for their excellent technical assistance. We also thank Enago (www.enago.jp) for the English language review.

Abbreviations

ADP

Adenosine 5′-diphosphate

ANOVA

Analysis of variance

ATP

Adenosine 5′-triphosphate

COX

Cyclooxygenase

EDCF

Endothelium-derived contracting factor

EDHF

Endothelium-derived hyperpolarizing factor

EDRFs

Endothelium-derived relaxing factors

GPCR

G protein-coupled receptor

KHS

Krebs–Henseleit solution

L-NNA

NG-nitro-L-arginine

NO

Nitric oxide

NOS

Nitric oxide synthase

PE

Phenylephrine

SHR

Spontaneously hypertensive rat

UDP

Uridine 5′-diphosphate

Up4A

Uridine adenosine tetraphosphate

UTP

Uridine 5′-triphosphate

VSMC

Vascular smooth muscle cell

WKY

Wistar Kyoto

Funding

This study was supported in part by JSPS KAKENHI Grant Numbers JP18K06861 (to Takayuki Matsumoto), JP17K08318 (to Kumiko Taguchi), and JP18K06974 (to Tsuneo Kobayashi) and The Promotion and Mutual Aid Corporation for Private Schools of Japan.

Compliance with ethical standards

Conflict of interest

Takayuki Matsumoto declares that he has no conflict of interest. Mihoka Kojima declares that she has no conflict of interest. Keisuke Takayanagi declares that he has no conflict of interest. Tomoki Katome declares that he has no conflict of interest. Kumiko Taguchi declares that she has no conflict of interest. Tsuneo Kobayashi declares that he has no conflict of interest.

Footnotes

Publisher’s note

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Contributor Information

Takayuki Matsumoto, Email: t-matsu@hoshi.ac.jp.

Tsuneo Kobayashi, Email: tkoba@hoshi.ac.jp.

References

  • 1.Matsumoto T, Goulopoulou S, Taguchi K, Tostes RC, Kobayashi T. Constrictor prostanoids and uridine adenosine tetraphosphate: vascular mediators and therapeutic targets in hypertension and diabetes. Br J Pharmacol. 2015;172:3980–4001. doi: 10.1111/bph.13205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 2.Burnstock G. Purinergic signaling in the cardiovascular system. Circ Res. 2017;120:207–228. doi: 10.1161/CIRCRESAHA.116.309726. [DOI] [PubMed] [Google Scholar]
  • 3.Burnstock G, Ralevic V. Purinergic signaling and blood vessels in health and disease. Pharmacol Rev. 2014;66:102–192. doi: 10.1124/pr.113.008029. [DOI] [PubMed] [Google Scholar]
  • 4.Erlinge D, Burnstock G. P2 receptors in cardiovascular regulation and disease. Prinergic Signal. 2008;4:1–20. doi: 10.1007/s11302-007-9078-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5.von Kügelgen I, Hoffmann K. Pharmacology and structure of P2Y receptors. Neuropharmacology. 2016;104:50–61. doi: 10.1016/j.neuropharm.2015.10.030. [DOI] [PubMed] [Google Scholar]
  • 6.Abbracchio MP, Burnstock G, Boeyaems JM, Barnard EA, Boyer JL, Kennedy C, Knight GE, Fumagalli M, Gachet C, Jacobson KA, Weisman GA. International Union of Pharmacology LVIII: update on the P2Y G protein-coupled nucleotide receptors: from molecular mechanisms and pathophysiology to therapy. Pharmacol Rev. 2006;58:281–341. doi: 10.1124/pr.58.3.3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7.Wihlborg AK, Balongh J, Wang L, Borna C, Dou Y, Joshi BV, Lazarowski E, Jacobson KA, Arner A, Erlinge D. Positive inotropic effects by uridine triphosphate (UTP) and uridine diphosphate (UDP) via P2Y2 and P2Y6 receptors on cardiomyocytes and release of UTP in man during myocardial infarction. Circ Res. 2006;98:970–976. doi: 10.1161/01.RES.0000217402.73402.cd. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8.Gergs U, Simm A, Bushnaq H, Silber RE, Neumann J. A positive inotropic effect of UTP in the human cardiac atrium. Eur J Pharmacol. 2014;724:24–30. doi: 10.1016/j.ejphar.2013.12.022. [DOI] [PubMed] [Google Scholar]
  • 9.Braun OO, Lu D, Aroonsakool N, Insel PA. Uridine triphosphate (UTP) induces profibrotic responses in cardiac fibroblasts by activation of P2Y2 receptors. J Mol Cell Cardiol. 2010;49:362–369. doi: 10.1016/j.yjmcc.2010.05.001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Morris JB, Pham TM, Kenney B, Sheppard KE, Woodcock EA. UTP transactivates epidermal growth factor receptors and promotes cardiomyocyte hypertrophy despite inhibiting transcription of the hypertrophic marker gene, atrial natriuretic peptide. J Biol Chem. 2004;279:8740–8746. doi: 10.1074/jbc.M310012200. [DOI] [PubMed] [Google Scholar]
  • 11.Patel JJ, Zhu D, Opdebeeck B, D’Haese P, Millan JL, Bourne LE, Wheeler-Jones CPD, Arnett TR, MacRae VE, Orriss IR. Inhibition of arterial medial calcification and bone mineralization by extracellular nucleotides: the same functional effect mediated by different cellular mechanisms. J Cell Physiol. 2018;233:3230–3243. doi: 10.1002/jcp.26166. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12.Pillois X, Chaulet H, Belloc I, Dupuch F, Desgranges C, Gadeau AP. Nucleotide receptors involved in UTP-induced rat arterial smooth muscle migration. Circ Res. 2002;90:678–681. doi: 10.1161/01.res.0000013700.98464.8e. [DOI] [PubMed] [Google Scholar]
  • 13.Jankowski V, Tolle M, Vanholder R, Schonfelder G, van der Giet M, Henning L, Schluter H, Paul M, Zidek W, Jankowski J. Uridine adenosine tetraphosphate: a novel endothelium-derived vasoconstrictive factor. Nat Med. 2005;11:223–227. doi: 10.1038/nm1188. [DOI] [PubMed] [Google Scholar]
  • 14.Ishida K, Matsumoto T, Taguchi K, Kamata K, Kobayashi T. Mechanisms underlying altered extracellular nucleotide-induced contractions in mesenteric arteries from rats in later-stage type 2 diabetes: effect of ANG II type 1 receptor antagonism. Am J Physiol Heart Circ Physiol. 2011;301:H1850–H1861. doi: 10.1152/ajpheart.00502.2011. [DOI] [PubMed] [Google Scholar]
  • 15.Winter P, Dora KA. Spreading dilatation to luminal perfusion of ATP and UTP in rat isolated small mesenteric arteries. J Physiol. 2007;582:335–347. doi: 10.1113/jphysiol.2007.135202. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16.Guns PJ, Korda A, Crauwels HM, Van Assche T, Robaye B, Boeynaems JM, Bult H. Pharmacological characterization of nucleotide P2Y receptors on endothelial cells of the mouse aorta. Br J Pharmacol. 2005;146:288–295. doi: 10.1038/sj.bjp.0706326. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17.You J, Golding EM, Bryan RM., Jr Arachidonic acid metabolites, hydrogen peroxide, and EDHF in cerebral arteries. Am J Physiol Heart Circ Physiol. 2005;289:H1077–H1083. doi: 10.1152/ajpheart.01046.2004. [DOI] [PubMed] [Google Scholar]
  • 18.Malmsjø M, Chu ZM, Croft K, Erlinge D, Edvinsson L, Beilin LJ. P2Y receptor-induced EDHF vasodilatation is of primary importance for the regulation of perfusion pressure in the peripheral circulation of the rat. Acta Physiol Scand. 2002;174:301–309. doi: 10.1046/j.1365-201x.2002.00956.x. [DOI] [PubMed] [Google Scholar]
  • 19.Malmsjo M, Hou M, Harden TK, Pendergast W, Pantev E, Edvinsson L, Erlinge D. Characterization of contractile P2 receptors in human coronary arteries by use of the stable pyrimidines uridine 5’-O-thiodiphosphate and uridine 5’-O-3-thiotriphosphate. J Pharmacol Exp Ther. 2000;293:755–760. [PubMed] [Google Scholar]
  • 20.Wihlborg AK, Malmsjo M, Eyjolfsson A, Gustafsson R, Jacobson K, Erlinge D. Extracellular nucleotides induce vasodilatation in human arteries via prostaglandins, nitric oxide and endothelium-derived hyperpolarizing factor. Br J Pharmacol. 2003;138:1451–1458. doi: 10.1038/sj.bjp.0705186. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21.Vanhoutte PM, Shimokawa H, Feletou M, Tang EH. Endothelial dysfunction and vascular disease – a 30th anniversary update. Acta Physiol. 2017;219:22–96. doi: 10.1111/apha.12646. [DOI] [PubMed] [Google Scholar]
  • 22.Feletou M, Verbeuron TJ, Vanhoutte PM. Endothelium-dependent contractions in SHR: a tale of prostanoid TP and IP receptors. Br J Pharmacol. 2009;156:563–574. doi: 10.1111/j.1476-5381.2008.00060.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23.Fisher SA. Vascular smooth muscle phenotype diversity and function. Physiol Genomics. 2010;42A:169–187. doi: 10.1152/physiolgenomics.00111.2010. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24.Dominiczak AF, Bohr DF. Vascular smooth muscle in hypertension. J Hypertens Suppl. 1989;7:S107–574S15. doi: 10.1097/00004872-198911000-00035. [DOI] [PubMed] [Google Scholar]
  • 25.De Silva TM, Kinzenbau DA, Modrick ML, Reinhardt LD, Faraci FM. Heterogenous impact of ROCK2 on carotid and cerebrovascular function. Hypertension. 2016;68:809–817. doi: 10.1161/HYPERTENSIONAHA.116.07430. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 26.Denniss SG, Ford RJ, Smith CS, Jeffery AJ, Rush JW. Chronic in vivo or acute in vitro resveratrol attenuates endothelium-dependent cyclooxygenase-mediated contractile signaling in hypertensive rat carotid artery. J Appl Physiol. 2016;120:1141–1150. doi: 10.1152/japplphysiol.00675.2015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 27.Hongo K, Nakagomi T, Kassell NF, Sasaki T, Lehman M, Vollmer DG, Tsukahara T, Ogawa H, Torner J. Effects of aging and hypertension on endothelium-dependent vascular relaxation in rat carotid artery. Stroke. 1988;19:892–897. doi: 10.1007/s00424-016-1834-5. [DOI] [PubMed] [Google Scholar]
  • 28.Watanabe S, Matsumoto T, Ando M, Adachi T, Kobayashi S, Iguchi M, Takeuchi M, Taguchi K, Kobayashi T. Multiple activation mechanisms of serotonin-mediated contraction in the carotid arteries obtained from spontaneously hypertensive rats. Pflugers Arch. 2016;468:1271–1282. doi: 10.1007/s00424-016-1834-5. [DOI] [PubMed] [Google Scholar]
  • 29.Mukohda M, Okada M, Hara Y, Yamawaki H. Methylglyoxal accumulation in arterial walls causes vascular contractile dysfunction in spontaneously hypertensive rats. J Pharmacol Sci. 2012;120:26–35. doi: 10.1254/jphs.12088fp. [DOI] [PubMed] [Google Scholar]
  • 30.Matsumoto T, Noguchi E, Ishida K, Kobayashi T, Yamada N, Kamata K. Metformin normalizes endothelial function by suppressing vasoconstrictor prostanoids in mesenteric arteries from OLETF rats, a model of type 2 diabetes. Am J Physiol Heart Circ Physiol. 2008;295:H1165–H1176. doi: 10.1152/ajpheart.00486.2008. [DOI] [PubMed] [Google Scholar]
  • 31.Matsumoto T, Nakayama N, Ishida K, Kobayashi T, Kamata K. Eicosapentaenoic acid improves imbalance between vasodilator and vasoconstrictor actions of endothelium-derived factors in mesenteric arteries from rat at chronic stage of type 2 diabetes. J Pharmacol Exp Ther. 2009;329:324–334. doi: 10.1124/jpet.108.148718. [DOI] [PubMed] [Google Scholar]
  • 32.Matsumoto T, Watanabe S, Kawamura R, Taguchi K, Kobayashi T. Epigallocatechin gallate attenuates ET-1-induced contraction in carotid artery from type 2 diabetic OLETF rat at chronic stage of disease. Life Sci. 2014;118:200–205. doi: 10.1016/j.lfs.2013.11.016. [DOI] [PubMed] [Google Scholar]
  • 33.Matsumoto T, Watanabe S, Yamada K, Ando M, Iguchi M, Taguchi K, Kobayashi T. Relaxation induced by atrial natriuretic peptide is impaired in carotid but not renal arteries from spontaneously hypertensive rats due to reduced BKCa channel activity. Biol Pharm Bull. 2015;38:1801–1808. doi: 10.1248/bpb.b15-00527. [DOI] [PubMed] [Google Scholar]
  • 34.Matsumoto T, Watanabe S, Iguchi M, Ando M, Oda M, Nagata M, Yamada K, Taguchi K, Kobayashi T. Mechanisms underlying enhanced noradrenaline-induced femoral arterial contractions of spontaneously hypertensive rats: involvement of endothelium-derived factors and cyclooxygenase-derived prostanoids. Biol Pharm Bull. 2016;39:384–393. doi: 10.1248/bpb.b15-00821. [DOI] [PubMed] [Google Scholar]
  • 35.Matsumoto T, Kojima M, Takayanagi K, Katome T, Taguchi K, Kobayashi T. Amplification of the COX/TXS/TP receptor pathway enhances uridine diphosphate-induced contraction by advanced glycation end products in rat carotid arteries. Pflugers Arch. 2019;471:1505–1517. doi: 10.1007/s00424-019-02330-y. [DOI] [PubMed] [Google Scholar]
  • 36.Haanes KA, Spray S, Syberg S, Jorgensen NR, Robaye B, Boeynaems JM, Edvinsson L. New insights on pyrimidine signalling within the arterial vasculature – different roles for P2Y2 and P2Y6 receptors in large and small coronary arteries of the mouse. J Mol Cell Cirdiol. 2016;93:1–11. doi: 10.1016/j.yjmcc.2016.01.025. [DOI] [PubMed] [Google Scholar]
  • 37.Guns PJ, Van Assche T, Fransen P, Robaye B, Boeynaems JM, Bult H. Endothelium-dependent relaxation evoked by ATP and UTP in the aorta of P2Y2-deficient mice. Br J Pharmacol. 2006;147:569–574. doi: 10.1038/sj.bjp.0706642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 38.Watanabe S, Matsumoto T, Ando M, Kobayashi S, Iguchi M, Taguchi K, Kobayashi T. A comparative study of Vasorelaxant effects of ATP, ADP, and adenosine on the superior mesenteric artery of SHR. Biol Pharm Bull. 2016;39:1374–1380. doi: 10.1248/bpb.b16-00260. [DOI] [PubMed] [Google Scholar]
  • 39.Matsumoto T, Tostes RC, Webb RC. Uridine adenosine tetraphosphate-induced contraction is increased in renal but not pulmonary arteries from DOCA-salt hypertensive rats. Am J Physiol Heart Circ Physiol. 2011;301:H409–H417. doi: 10.1152/ajpheart.00084.2011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 40.Matsumoto T, Tostes RC, Webb RC. Alterations in vasoconstrictor responses to the endothelium-derived contracting factor uridine adenosine tetraphosphate are region specific in DOCA-salt hypertensive rats. Pharmacol Res. 2012;65:81–90. doi: 10.1016/j.phrs.2011.09.005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 41.Alsaqati M, Chan SL, Ralevic V. Investigation of the functional expression of purine and pyrimidine receptors in porcine isolated pancreatic arteries. Purinergic Signal. 2014;10:241–249. doi: 10.1007/s11302-013-9403-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 42.Malmsjö M, Bergdahl A, Zhao XH, Sun XY, Hedner T, Edvinsson L, Erlinge D. Enhanced acetylcholine and P2Y-receptor stimulated vascular EDHF-dilatation in congestive heart failure. Cardiovasc Res. 1999;43:200–209. doi: 10.1016/s0008-6363(99)00062-0. [DOI] [PubMed] [Google Scholar]
  • 43.Marrelli SP. Mechanisms of endothelial P2Y(1)- and P2Y(2)-mediated vasodilatation involve differential [Ca2+]i responses. Am J Physiol Heart Circ Physiol. 2001;281:H1759–H1766. doi: 10.1152/ajpheart.2001.281.4.H1759. [DOI] [PubMed] [Google Scholar]
  • 44.Gluais P, Lonchampt M, Morrow JD, Vanhoutte PM, Feletou M. Acetylcholine-induced endothelium-dependent contractions in the SHR aorta: the Janus face of prostacyclin. Br J Pharmacol. 2005;146:834–845. doi: 10.1038/sj.bjp.0706390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 45.Gluais P, Paysant J, Badier-Commander C, Verbeuren T, Vanhoutte PM, Feletou M. In SHR aorta, calcium ionophore A-23187 releases prostacyclin and thoromboxane A2 as endothelium-derived contracting factors. Am J Physiol Heart Circ Physiol. 2006;291:H2255–H2264. doi: 10.1152/ajpheart.01115.2005. [DOI] [PubMed] [Google Scholar]
  • 46.Spitler KM, Matsumoto T, Webb RC. Suppression of endoplasmic reticulum stress improves endothelium-dependent contractile responses in aorta of the spontaneously hypertensive rat. Am J Physiol Heart Circ Physiol. 2013;305:H344–H353. doi: 10.1152/ajpheart.00952.2012. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 47.Lüscher TF, Aarhus LL, Vanhoutte PM. Indomethacin improves the impaired endothelium-dependent relaxations in small mesenteric arteries of the spontaneously hypertensive rat. Am J Hypertens. 1990;3:55–58. doi: 10.1093/ajh/3.1.55. [DOI] [PubMed] [Google Scholar]
  • 48.Horiuchi T, Dietrich HH, Tsugane S, Dacey RG., Jr Analysis of purine- and pyrimidine-induced vascular responses in the isolated rat cerebral arteriole. Am J Physiol Heart Circ Physiol. 2001;280:H767–H776. doi: 10.1152/ajpheart.2001.280.2.H767. [DOI] [PubMed] [Google Scholar]
  • 49.Li Y, Baylie RL, Tavares MJ, Brayden JE. TRPM4 channels couple purinergic receptor mechanoactivation and myogenic tone development in cerebral parenchymal arterioles. J Cereb Blood Flow Metab. 2014;34:1706–1714. doi: 10.1038/jcbfm.2014.139. [DOI] [PMC free article] [PubMed] [Google Scholar]

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