pituitrin and dazoxiben

Estrogen potentiates vascular reactivity to vasopressin (VP) by enhancing constrictor prostanoid function. To determine the cellular and molecular mechanisms, the effects of estrogen on arachidonic acid metabolism and on the expression of constrictor prostanoid pathway enzymes and endoperoxide/thromboxane receptor (TP) were determined in the female rat aorta. The release of thromboxane A2 (TxA2) and prostacyclin (PGI2) was measured in male (M), intact-female (Int-F), ovariectomized-female (OvX-F), and OvX + 17beta-estradiol-replaced female (OvX + ER-F) rats. The expression of mRNA for cyclooxygenase (COX)-1, COX-2, thromboxane synthase (TxS), and TP by aortic endothelium (Endo) and vascular smooth muscle (VSM) of these four experimental groups was measured by RT-PCR. The expression of COX-1, COX-2, and TxS proteins by Endo and VSM was also estimated by immunohistochemistry (IHC). Basal release of TxA2 and PGI2 was similar in M (18.8 +/- 1.9 and 1,723 +/- 153 pg/mg ring wt/45 min, respectively) and Int-F (20.2 +/- 4.2 and 1,488 +/- 123 pg, respectively) rat aortas. VP stimulated the dose-dependent release of TxA2 and PGI2 from both male and female rat aorta. OvX markedly attenuated and ER therapy restored VP-stimulated release of TxA2 and PGI2 in female rats. No differences in COX-1 mRNA levels were detected in either Endo or VSM of the four experimental groups (P > 0.1). The expression of both COX-2 and TxS mRNA were significantly higher (P < 0.05) in both Endo and VSM of Int-F and OvX + ER-F, compared with M or OvX-F. Expression of TP mRNA was significantly higher in VSM of Int-F and OvX + ER-F compared with M or OvX-F. IHC revealed the uniform staining of COX-1 in VSM of the four experimental groups, whereas staining of COX-2 and TxS was greater in Endo and VSM of Int-F and OvX + ER-F than in OvX-F or M rats. These data reveal that estrogen enhances constrictor prostanoid function in female rat aorta by upregulating the expression of COX-2 and TxS in both Endo and VSM and by upregulating the expression of TP in VSM.

Topics: Animals; Aorta, Thoracic; Cyclooxygenase 1; Cyclooxygenase 2; Drug Implants; Endothelium, Vascular; Enzyme Induction; Enzyme Inhibitors; Epoprostenol; Estradiol; Female; Imidazoles; Immunohistochemistry; Male; Membrane Proteins; Muscle, Smooth, Vascular; Ovariectomy; Radioimmunoassay; Rats; Rats, Sprague-Dawley; Receptors, Thromboxane; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Signal Transduction; Thromboxane A2; Thromboxane-A Synthase; Up-Regulation; Vasoconstriction; Vasopressins

The effects of constrictor prostanoid (CP) pathway inhibitors on vascular reactivity to vasopressin (VP) and phenylephrine (PE) were examined in thoracic aortas of male, female, and ovariectomized (OVX) female Sprague-Dawley rats. Maximal contractile response of control (Cont) aortas to VP was markedly higher in females (3,885 +/- 332 mg/mg ring wt) than in males (810 +/- 148 mg). Indomethacin (Indo; 10 microM) attenuated maximal response to VP in females (3,043 +/- 277 mg) but not in males. SQ-29,548 (SQ; 1 microM) attenuated maximal response to VP in females (3,042 +/- 290 mg) to a similar extent as Indo. Dazoxiben (Daz; 10 microM) alone had no effect, but Daz + SQ attenuated maximal contractile response to VP to a similar extent as SQ alone. Removal of the endothelium in female aortas attenuated contractile responses to VP in Cont aortas. OVX attenuated maximal contractile response to VP in Cont aortas (2,093 +/- 329 mg) and abolished the attenuating effects of Indo. Indo, SQ, and Daz exerted identical effects on contractile responses of male, female, and OVX female aortas to PE. These findings establish the following in the rat aorta: 1) CP, probably thromboxane and/or endoperoxide, is responsible for approximately 25-30% of contractile responses of females, but not males, to VP and PE; 2) CP production by the female aorta is primarily endothelial in origin; and 3) ovarian steroids modulate production and/or actions of CP in female aortas.

Topics: Animals; Aorta; Bridged Bicyclo Compounds, Heterocyclic; Cyclooxygenase Inhibitors; Endothelium, Vascular; Enzyme Inhibitors; Estrogens; Fatty Acids, Unsaturated; Female; Hydrazines; Imidazoles; Indomethacin; Male; Ovariectomy; Phenylephrine; Progesterone; Prostaglandins; Rats; Rats, Sprague-Dawley; Sex Characteristics; Thromboxanes; Vasoconstriction; Vasoconstrictor Agents; Vasopressins

Sympathetic neurotransmission can be modulated by prostaglandins in a number of tissues, but it is not known whether thromboxane A2 also influences neurotransmission. In this study, vasoconstrictor responses to electrical stimulation of the sympathetic nerves and to injection of noradrenaline were examined in the blood perfused mesentery of the rat in situ. The thromboxane synthetase inhibitor dazoxiben, infused into the perfusion circuit at 10-100 mumol/l, significantly inhibited constrictor responses to nerve stimulation and to injected noradrenaline and vasopressin. The cyclo-oxygenase inhibitor indomethacin (28 mumol/kg intravenously) had no effect on responses to nerve stimulation or noradrenaline, but pretreatment with indomethacin abolished the inhibitory effect of dazoxiben on vasoconstrictor responses. The thromboxane-mimetic (U46619, 10 nmol/l) slightly reduced responses to nerve stimulation (but not to noradrenaline), whereas prostacyclin (3-10 nmol/l) and PGE2 (3 nmol/l) markedly reduced responses both to nerve stimulation and to injections of noradrenaline. These prostanoids did not alter perfusion pressure at these concentrations. The data suggest that the inhibitory effect of dazoxiben on sympathetic neurotransmission is unlikely to be due directly to inhibition of thromboxane synthesis. Inhibition might result from diversion of endoperoxide metabolism to endogenous prostanoids that, in turn, inhibit activation of vasoconstrictor mechanisms.

Topics: Animals; Cyclooxygenase Inhibitors; Electric Stimulation; Imidazoles; Indomethacin; Male; Norepinephrine; Oxidoreductases; Prazosin; Prostaglandins; Rats; Rats, Inbred Strains; Splanchnic Circulation; Sympathetic Nervous System; Thromboxane-A Synthase; Vasoconstriction; Vasopressins