Thromboxane A2 Receptor Density Research Articles

SEX SPECIFIC CHANGES OF CAROTID ARTERIAL FUNCTION IN RESPONSE TO VITAMIN D DEFICIENCY

Objective: Male sex and Vitamin D deficiency is associated with increased ischemic stroke risk that may be partially explained by local changes of circulatory control. Our aim was to examine the vascular reactivity and histological changes of isolated carotid artery of female and male rats in Vitamin D deficient and Vitamin D supplemented state. Design and method: Female and male Wistar rats were fed with vitamin D reduced rat chow for eight weeks. Another group received normal chow with further supplementation of vitamin D. Phenylephrine induced contraction, acetylcholine and estrogen induced relaxation of carotid arteries was examined by wire myography. Elastic fiber density was evaluated on resorcin-fuchsin stained tissue sections. Smooth muscle cell actin (SMA), thromboxane A2 receptor and endothelial nitric oxide synthase (eNOS) density and protein tyrosine nitration (NT) was measured by immunohistochemistry. Results: Both male sex and vitamin D deficiency resulted in increased phenylephrine induced contraction. Inhibition of cyclooxygenases by indomethacin increased the degree of contraction only in vitamin D supplemented females. Male gender was associated with reduced acetylcholine induced relaxation, and it was not altered by vitamin D deficiency. Indomethacin increased relaxation in male vessels. Estrogen induced relaxation was significantly reduced by vitamin D deficiency in female but not in male animals. Vitamin D deficiency resulted in reduced elastic fiber density in female, but decreased smooth muscle cell actin density in male rats. Thromboxane A2 receptor density was increased in vitamin D deficient males. NT immunohistochemistry was low in both male group, whereas eNOS was reduced in vitamin D deficient males. Conclusions: In rats male gender was associated with increased phenylephrine induced contraction and reduced acetylcholine induced relaxation of the carotid artery. Vitamin D deficiency caused increased contraction in both sexes; did not change acetylcholine induced relaxation, but reduced the estrogen induced relaxation in female arteries. These functional differences were accompanied with sex specific histological changes of the vessel. The observed sex specific properties and responses of carotid artery to vitamin D deficiency may play a role in the gender difference of cardiovascular and stroke risk.

Corrigendum

CORRIGENDACorrigendumPublished Online:01 Dec 2005https://doi.org/10.1152/ajpheart.01029.2005Original articleMoreSectionsPDF (27 KB)Download PDF ToolsExport citationAdd to favoritesGet permissionsTrack citations ShareShare onFacebookTwitterLinkedInEmailWeChat Volume 289, August 2005 Volume 58, August 2005 Pages H578–H585: R. J. Gonzales, A. A. Ghaffari, S. P. Duckles, and D. N. Krause. “Testosterone treatment increases thromboxane function in rat cerebral arteries.” Reference 1 in this paper was incorrectly cited. It should appear as the following:Ajayi AA, Mathur R, and Halushka PV. Testosterone increases human platelet thromboxane A2 receptor density and aggregation responses. Circulation 91: 2742–2747, 1995.This article has no references to display. Download PDF Previous Back to Top FiguresReferencesRelatedInformationRelated ArticlesTestosterone treatment increases thromboxane function in rat cerebral arteries 01 Aug 2005American Journal of Physiology-Heart and Circulatory Physiology More from this issue > Volume 289Issue 6December 2005Pages H2754-H2754 Copyright & PermissionsCopyright © 2005 by the American Physiological Societyhttps://doi.org/10.1152/ajpheart.01029.2005History Published online 1 December 2005 Published in print 1 December 2005 Metrics

Castration reduces platelet thromboxane A2 receptor density and aggregability

Exogenously administered testosterone upregulates platelet thromboxane A2 (TXA2) receptors and increases aggregation response to thromboxane mimetics in healthy male volunteers. However, the biological impact of endogenous testosterone on platelet TXA2 receptor expression, especially in older men at risk of coronary artery disease, is unclear. To investigate the impact of reduction in circulating testosterone on platelet TXA2 receptor expression in older men. Cross-sectional case-control study. We studied surgically and/or medically castrated men with prostate cancer (group A, n = 8, aged 71 +/- 8 years) and age-matched, uncastrated urology patients (group B, n = 7, aged 67 +/- 9 years). Plasma testosterone was measured by radioimmunoassay. Platelet TXA2 receptor expression was assessed by radioligand binding studies using radioactive 125I-BOP. Platelet aggregation responses to TXA2-mimetic I-BOP, and to thrombin, were also studied. Group A had significantly lower plasma testosterone than group B (16 +/- 5 ng/dl vs. 308 +/- 47 ng/dl, p<0.001). Platelet TXA2 receptor density (B(max)) but not affinity (K(d)) was lower in group A (0.50 +/- 0.12 vs. 1.01 +/- 0.17 pmol/mg protein, p = 0.03). Maximum platelet aggregation response to I-BOP (E(max)), but not sensitivity (EC50) was lower in group A (53 +/- 2% vs. 63 +/- 2%, p = 0.003 ANOVA). In vitro, high concentrations of hydroxyflutamide (100 microM) competitively inhibited U46619-induced platelet aggregation in washed platelets, without affecting the binding of 125I-BOP to platelet TXA2 receptors. Endogenous testosterone regulates platelet TXA2 receptor B(max) and the E(max) aggregation response to thromboxane mimetic I-BOP. Blockade of androgen receptors or inhibition of testosterone production may reduce platelet aggregation responses. Preliminary evidence suggests the presence of functional androgen receptors on human platelets, which may regulate TXA2 receptor expression.

Gender-Related Differences in Androgen Regulation of Thromboxane A2 Receptors in Rat Aortic Smooth-Muscle Cells

Thromboxane A2 (TXA2) has been implicated as an important mediator of cardiovascular diseases. Aortas obtained from male rats are more sensitive to TXA2 mimetics compared with those obtained from females. A similar phenomenon has been reported in canine coronary arteries. To determine whether there is a gender-related difference in the regulation of TXA2 receptors by androgenic steroids, we determined the effect of testosterone and dihydrotestosterone (DHT) on TXA2 receptor density in cultured rat aortic smooth-muscle (RASM) cells and guinea pig coronary artery smooth-muscle (CASM) cells. TXA2 receptor density (B(max)) and dissociation constant (Kd) were determined by radioligand binding studies with (125)I-BOP, a TXA2 receptor agonist. Testosterone significantly (p < 0.05) increased TXA2 receptor density in cultured RASM cells and guinea pig CASM cells. DHT significantly (p < 0.005) increased the B(max) in male RASM cells (62 +/- 2 vs. 40 +/- 3 fmol/mg protein; n = 7; p < 0.005). DHT increased the B(max) values in both male and female RASM cells, but the increase was significantly (p < 0.05) less in female than in male RASM cells (57 +/- 10% increase for male and 31 +/- 5% for female). Androgen-receptor protein was detected in RASM cells by Western blot and was less in the female RASM cells than in the male. The results indicate that RASM cells possess an androgen receptor and that gender-related differences exist in the regulation of expression of TXA2 receptors by androgens.

Thromboxane A2 Receptor Density Increases during Chronic Exposure to Thromboxane A2 Receptor Antagonists after Porcine Carotid Bypass

Domestic swine ( n = 12 in each group) were randomized to daily treatment with the thromboxane A2 (TXA2) receptor antagonist BMS-180291 (group I), aspirin (group II), or no drug (group III) prior to prosthetic carotid graft implantation. Platelet and arterial wall receptor density were measured by equilibrium binding using 125I-BOP. At 6 weeks, means (s.e.m.) platelet receptor density (pmol/mg) had increased in groups I (3.3(0.6) versus 1.8(0.3); P&lt;0.05) and II (2.6(0.6) versus 1.7(0.2); P&lt;0.05), but not in group III (1.3(0.3) versus 1.2(0.2)). Aortic membrane TXA2 receptor density (fmol/mg) was significantly greater ( P&lt;0.05) in groups I (150(50)) and II (68(10)) compared with group III (39(6)). Chronic exposure to a TXA2 receptor antagonist or aspirin is associated with increased platelet and aortic receptor density in pigs. Published by Elsevier Science Ltd.

Androstenedione increases thromboxane A2 receptors in human erythroleukemia cells

Previous studies have demonstrated an increased thromboxane A2 (TXA2) receptor expression in Human Erythroleukemia (HEL) cells and rat aortic smooth muscle (RASM) cells in response to testosterone treatment. HEL cells have served as a model for megakaryocytes, the progenitor cell for platelets. Platelets have previously been shown to convert androstenedione to testosterone. This study investigated the effects of androstenedione on the TXA2 receptor density in HEL and cultured RASM cells. Both cell lines were incubated with vehicle, 150 nM testosterone or 250, 500 or 750nM androstenedione for 48 hours. Co-incubation with testosterone or androstenedione significantly (p<0.05) increased the maximum number of TXA2 binding sites (Bmax) in HEL cells compared to controls. There was no significant change in Kd values. In a separate series of experiments, HEL cells were incubated with the androgen receptor antagonist hydroxyflutamide (2.5μM). Treatment with androstenedione (500nM) significantly (p<0.05) increased the Bmax value by 35% compared to control and hydroxyflutamide completely antagonized this effect of androstenedione. Incubation with hydroxyflutamide alone had no effect on the Bmax values compared to control. RASM cells also showed an increase in Bmax values by 25% and 23% over control (95±6.6, 118±7.2 and 117±5.1 fmoles/mg protein, control, testosterone and androstenedione, n=3). Both cell lines converted androstenedione to testosterone. The results raise the possibility that the adrenal androgen, androstenedione can regulate the expression of TXA2 receptors either on its own or via conversion to testosterone and through an androgen receptor.

Testosterone regulation of platelet and vascular thromboxane A2 receptors.

Testosterone has been implicated as a risk factor for cardiovascular diseases and thromboxane A2 (TXA2) plays a role in these diseases. We tested the notion that testosterone regulates the expression of TXA2 receptors in platelets and vascular smooth muscle. Testosterone significantly increased the density of TXA2 receptors in cultured rat aortic smooth muscle and human erythroleukemia cells, a megakaryocyte-like cell. Treatment of rats with testosterone resulted in a significant increase in platelet and aortic TXA2 receptor density and increased responsiveness to TXA2 mimetics. We conclude that testosterone regulates the expression of TXA2 receptors.

Testosterone increases thromboxane A2 receptor density and responsiveness in rat aortas and platelets

Testosterone has been implicated as a risk factor for cardiovascular diseases. Thromboxane (Tx) A2 is an important pathophysiological mediator for thrombotic vascular diseases. This study investigated the effects of testosterone on platelet and vascular TxA2 receptors. Male rats were treated with either testosterone cypionate for 2 wk, sham operated, castrated, or castrated and treated with testosterone cypionate for 2 wk. Treatment of intact rats with testosterone significantly (P < 0.001) increased the TxA2 receptor density in platelets from 25.4 +/- 3.2 to 42.9 +/- 4.2 fmol/mg protein (P < 0.005, n = 17) and in aortic membranes from 48.7 +/- 1.7 to 86.1 +/- 6.1 fmol/mg protein, n = 9. The threshold concentration of the TxA2 mimetic, [1S-(1 alpha, 2 beta(5Z),3 alpha(1E,3R*)4 alpha)]-7-[3-(3-hydroxy-4- (4'-iodophenoxy)-1-butenyl)-7-oxabicyclo[2.21]heptan-2-yl]-5 -heptenoic acid (I-BOP), to induce platelet aggregation was significantly (P < 0.01) decreased from 0.45 +/- 0.16 nM, n = 7, in the control rats to 0.07 +/- 0.01 nM, n = 13, in the testosterone-treated rats. Testosterone treatment resulted in a significantly (P < 0.05) greater maximum aortic contractile response to the TxA2 mimetic, U-46619, compared with intact rats. Castration resulted in a significant (P < 0.01) decrease in aortic TxA2 receptor density from 51.7 +/- 3.7 to 27.3 +/- 5.3 fmol/mg protein, which was significantly reversed by testosterone treatment (89.2 +/- 7.1 fmol/mg protein; n = 4). Castration resulted in a significantly (P < 0.05) lower maximal aortic contractile response that was reversed by treatment with testosterone. Castration did not significantly change platelet TxA2 receptor density.(ABSTRACT TRUNCATED AT 250 WORDS)