Inhibition of RNA Synthesis by Bradykinin Involves Both the B1and B2Receptor Subtypes

doi:10.1006/abbi.1996.0150

Archives of Biochemistry and Biophysics

Volume 328, Issue 1, 1 April 1996, Pages 115-121

https://doi.org/10.1006/abbi.1996.0150 Get rights and content

Abstract

The efficacy of angiotensin converting enzyme inhibitors in the treatment of heart disease is due in part to the accumulation of bradykinin (BK). Since BK can exert its effect by influencing cell proliferation, we chose to study the effect of BK on the growth of A10 vascular smooth muscle cells. Ligand binding studies to determine which BK receptor subtypes are present on A10 cells showed that both B₁and B₂receptors were present in approximately equal numbers. Examination of RNA synthesis demonstrated that BK inhibits uridine incorporation in a dose-dependent manner. This decrease in RNA synthesis was blocked by both B₁and B₂receptor antagonists, as well as by addition of indomethacin, a cyclooxygenase inhibitor. The latter result suggested that prostaglandins mediate the biological actions of BK. Consequently, we examined the direct effect of two prostaglandins, PGE₂and PGI₂(prostacyclin), on A10 cells. PGE₂caused a decrease in RNA synthesis, thus mimicking the effect of BK, while PGI₂did not. Therefore, the inhibition of RNA synthesis in A10 vascular smooth muscle cells by BK requires both B₁and B₂receptor subtypes and this action of BK is apparently mediated byde novosynthesis of prostaglandins.

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Cited by (15)

The role of the kallikrein-kinin system, matrix metalloproteinases, and tissue inhibitors of metalloproteinases in the early restenosis of covered stents in the femoropopliteal arterial segment
2017, Journal of Vascular Surgery
Citation Excerpt :
In contrast, when the endothelial cells were absent, BK had a direct effect on hypertrophy of the ventricular myocytes.27 Several studies have reported that BK inhibits aortic SMC proliferation in mouse models.28,29 This point was emphasized by Agata et al30 who reported that the increased production of kallikrein in rat carotid arteries in an adenovirus-mediated gene transfer model, which presumably resulted in increased synthesis of BK, reduced myointimal hyperplasia following balloon angioplasty.
The purpose of this study was to investigate the roles of the kallikrein-kinin system and matrix metalloproteinases (MMPs) in the development of arterial restenosis attributable to intimal hyperplasia in the femoropopliteal arteries.
This report describes a single-center prospective study of 27 patients with peripheral artery disease who required percutaneous transluminal angioplasty and stenting of the femoropopliteal segment using covered stent grafts. The blood concentrations of total and kininogen fractions were evaluated using immunoenzymatic methods. Plasma kallikrein was evaluated by the colorimetric method. Tissue kallikrein was evaluated by the spectrophotometric method. The activity of kininase II was measured by fluorometric analysis. Quantification of MMPs was performed by zymography, and tissue inhibitors of metalloproteinases were measured by enzyme-linked immunosorbent assay.
Four (15%) of the treated patients developed restenosis at the 6-month follow-up evaluation. These patients had significantly lower levels of high-molecular-weight kininogens (24 hours; P < .05) and low-molecular-weight kininogens (before, P < .05; 24 hours, P < .01; 6 months, P < .05) and lower levels of tissue inhibitor of metalloproteinases-2 (6 months; P < .05) than the patients without restenosis. The activity levels of plasma and tissue kallikrein, kininase II, and MMPs did not differ significantly between the patients with and without restenosis.
This study demonstrates an involvement of the kallikrein-kinin system in in-stent restenosis, although we could not confirm the participation of metalloproteinases in the restenosis process.
Suppressed angiogenesis in kininogen-deficiencies
2002, Laboratory Investigation
We investigated whether the kinin-generating system enhanced angiogenesis in chronic and proliferative granuloma and in tumor-surrounding stroma. In rat sponge implants, angiogenesis was gradually developed in normal Brown Norway Kitasato rats (BN-Ki). The development of angiogenesis was significantly suppressed in kininogen-deficient Brown Norway Katholiek rats (BN-Ka). The angiogenesis enhanced by basic fibroblast growth factor was also significantly less marked in BN-Ka than in BN-Ki. Naturally occurring angiogenesis was significantly suppressed by B₁ or B₂ antagonist. mRNA of vascular endothelial growth factor was more highly expressed in the granulation tissues in BN-Ki than in BN-Ka. Daily topical injections of aprotinin, but not of soy bean trypsin inhibitor, suppressed angiogenesis. Daily topical injections of low-molecular weight kininogen enhanced angiogenesis in BN-Ka. Topical injections of serum from BN-Ki, but not from BN-Ka, also facilitated angiogenesis in BN-Ka. FR190997, a nonpeptide mimic of bradykinin, promoted angiogenesis markedly, with concomitant increases in vascular endothelial growth factor mRNA. Angiogenesis in the granulation tissues around the implanted Millipore chambers containing Walker-256 cells was markedly more suppressed in BN-Ka than in BN-Ki. Our results suggest that endogenous kinin generated from the tissue kallikrein-kinin system enhances angiogenesis in chronic and proliferative granuloma and in the stroma surrounding a tumor. Thus, the agents for the kinin-generating system and/or kinin receptor signaling may become useful tools for controlling angiogenesis.
Detection of bradykinin B<inf>1</inf> receptors in rat aortic smooth muscle cells
2001, Biochemical Pharmacology
Citation Excerpt :
However, although hypotensive responses to des-Arg9-bradykinin have been reported in lipopolysaccharide-treated rats [24,25], the rat aortain vitro does not seem to respond to des-Arg9-bradykinin [2]. This suggests that the bradykinin B1 receptor may not be significantly expressed in the rat aorta, although bradykinin B1 receptor mRNA has been detectedex vivo in the aorta of lipopolysaccharide-treated rats [26] and functional effects ascribed to bradykinin B1 receptors have been described in a rat smooth muscle cell line [27]. In an attempt to determine whether functional bradykinin B1 receptors are indeed expressed in rat aortic cells, we studied bradykinin B1 receptor expression in rat aortic smooth muscle cells by binding experiments and functional responses.
¹The tritiated bradykinin B₁ receptor agonist [³H]des-Arg¹⁰-kallidin bound to a single class of high-affinity binding sites (K_d = 0.5 ± 0.16 nM; B_max = 15,000 ± 8,000 sites/cell) on cultured rat aortic smooth muscle cells. [³H]Des-Arg¹⁰-kallidin association and dissociation kinetics were monoexponential, making it possible to determine the association and dissociation rate constants (k₊₁ = 1.5 10⁵ M⁻¹ sec⁻¹;k₋₁ = 4.2 10⁻⁵ sec⁻¹). [³H]Des-Arg¹⁰-kallidin binding was inhibited by specific ligands of bradykinin B₁ and B₂ receptors with a rank order of potency consistent with that known for bradykinin B₁ receptors in other species (des-Arg⁹-[Leu⁸]bradykinin = des-Arg¹⁰-kallidin = des-Arg⁹-bradykinin = des-Arg¹⁰-[Leu⁹]kallidin > des-Arg¹⁰-HOE-140≫bradykinin≫HOE-140). Bradykinin B₁ receptor mRNA was also detected in these cells. Des-Arg¹⁰-kallidin increased cytosolic free Ca²⁺ levels, phosphoinositide turnover, and arachidonic acid release at nanomolar concentrations (respective ec₅₀ values: 16 ± 2, 4 ± 2.7, 6 ± 2 nM). These functional effects of des-Arg¹⁰-kallidin could be blocked by the bradykinin B₁ receptor antagonist des-Arg⁹-[Leu⁸]bradykinin, but were not sensitive to bradykinin B₂ receptor antagonists. These results therefore show that rat aortic smooth muscle cells in culture express functional bradykinin B₁ receptors.
Sarpogrelate inhibits serotonin-induced proliferation of porcine coronary artery smooth muscle cells: Implications for long-term graft patency
2001, Annals of Thoracic Surgery
Citation Excerpt :
Multiple 5-HT receptor subtypes could potentially be involved in the induction of SMC proliferation. Other growth factors have already been demonstrated to exert their mitogenic effect through multiple receptors [27, 28]. The rationale for testing sarpogrelate under conditions of Ang II, endothelin, or PDGF stimulation was to determine whether 5-HT is a component of the mitogenic pathways associated with other growth factors, as has been proposed for PDGF and epidermal growth factor [22].
Background. Serotonin can induce proliferation of vascular smooth muscle cells. We assessed the ability of a specific serotonin receptor antagonist, sarpogrelate, to inhibit proliferation of cultured porcine coronary artery smooth muscle cells.
Methods. Cell proliferation and mitotic activity were measured using 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide. To determine the effect of sarpogrelate on DNA (deoxyribonucleic acid), RNA (ribonucleic acid), and protein synthesis, radioactive incorporation of ³H-thymidine, ³H-uridine, and ³H-phenylalanine, respectively, was used. Synthesis of DNA was also assessed by flow cytometry with propidium iodide as a fluorochrome.
Results. Serotonin, platelet-derived growth factor, endothelin, and angiotensin II all induced proliferation of porcine coronary artery smooth muscle cells. Sarpogrelate specifically inhibited the serotonin-induced cytokine trigger but did not influence platelet-derived growth factor–, endothelin–, or angiotensin II–induced cell proliferation. Sarpogrelate inhibited the serotonin-induced increase in intracellular free ionized calcium concentration, prevented mitogen-activated protein kinase activation, and down-regulated expression of the protooncogenes c-fos and c-jun. Sarpogrelate acted at the G₁ phase of the cell cycle.
Conclusions. These data suggest that sarpogrelate could be used as a therapeutic agent to inhibit serotonin-induced neointimal hyperplasia and improve patency of coronary artery bypass grafts.
Regulation of cardiac fibroblast extracellular matrix production by bradykinin and nitric oxide
1999, Journal of Molecular and Cellular Cardiology
The beneficial effects of angiotensin-converting enzyme inhibitors on ameliorating cardiac fibrosis have been partially attributed to their ability to prevent the degradation of kinins. The potential role of bradykinin and the related signaling molecule nitric oxide (NO) in modulating extracellular matrix (ECM) production was examined in primary cultures of adult rat cardiac fibroblasts. Treatment of fibroblasts with 5 n $m$ bradykinin for 24 h led to a reduction in steady-state mRNA levels for fibronectin (34±7%) and collagens type I (19±8%) and type III (48±4%), as determined by Northern blot analysis. The NO synthase inhibitor $l$ -NAME attenuated the reduction observed in fibronectin and collagen mRNA levels in response to bradykinin. The NO donor DETA NONOate (100 μ $m$ ) mimicked the effects of bradykinin on ECM mRNA levels. Protein levels of soluble fibronectin, assessed in conditioned medium by ELISA, were decreased by 14±4% and 21±4% after 48 h treatment with 1 μ $m$ bradykinin and 100 μ $m$ DETA NONOate, respectively. Bradykinin stimulated intracellular cGMP accumulation 73.7±10.3% after 10 min of treatment. Cell proliferation rates at 48 h were unaffected by bradykinin, but were reduced by 26±12% by 100 μ $m$ DETA NONOate. These data indicate that bradykinin downregulates ECM protein production in cardiac fibroblasts and suggest that NO and the related signaling molecule cGMP may play an important role in mediating this reponse.
Molecular cloning and expression of rat bradykinin B<inf>1</inf> receptor
1998, Biochimica et Biophysica Acta - Gene Structure and Expression
The gene encoding rat bradykinin B₁ receptor has been cloned by using a partial rat B₁ cDNA probe. The rat B₁ receptor gene contains two exons and the entire coding region is within the second exon. The 5′-flanking region of the rat B₁ receptor gene contains several putative transcriptional regulatory sites including TATA box, cAMP response element, NF-κB and AP-1. It showed promoter activity inducible by lipopolysaccharide in vascular smooth muscle cells. Rat B₁ receptor mRNA was found to be alternatively spliced and induced by lipopolysaccharide treatment in a wide range of tissues, such as the salivary gland, testis, kidney, lung, heart, prostate and aorta. The deduced rat B₁ receptor amino acid sequence is 71% homologous to human and rabbit counterparts, and 89% homologous to the mouse counterpart. The expressed B₁ receptor in HEK293 cells displayed a rank order of affinity for the kinin peptides: des-Arg⁹-BK>Lys-des-Arg⁹-BK≈des-Arg⁹,Leu⁸-BK>Sar-Tyr-ϵAhx-Lys-[d-βNal⁷,Ile⁸]-des-Arg⁹-BK>Sar-Tyr-ϵAhx-Lys-des-Arg⁹-BK≫BK≫Hoe140. These results indicate that the cloned gene encodes a functional rat B₁ receptor.

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Regular ArticleInhibition of RNA Synthesis by Bradykinin Involves Both the B1and B2Receptor Subtypes

Abstract

Regular Article
Inhibition of RNA Synthesis by Bradykinin Involves Both the B₁and B₂Receptor Subtypes