Penile constitutive nitric oxide synthase expression in rats exposed to unpredictable chronic mild stress: role of inflammation.

Chronic psychological stress cause erectile dysfunction (ED). Considering recent evidence that tumor necrosis factor-α (TNF-α) levels are increased in serum of patients with ED, the present study investigated the effects of infliximab (a TNF-α blocker) on endothelial nitric oxide synthase (eNOS) and neuronal NOS (nNOS) immunoreactivity of rat penile corpus cavernosum in unpredictable chronic mild stress (UCMS). Male adult rats were randomly divided into three groups (n=8 per group): Control, UCMS and UCMS+infliximab. Control and UCMS groups received physiological saline, UCMS+infliximab group received infliximab (5 mg kg-1 per week, intraperitoneally) during 8 weeks of UCMS. UCMS and UCMS+infliximab groups were subjected to different types of stressors, which were randomly applied four to five times during this time period. After 8 weeks, penile eNOS and nNOS expressions were determined immunohistochemically. In UCMS group, nNOS and eNOS immunoreactivity was found to be decreased in penile corpus cavernosum compared with the control group. Whereas in infliximab treatment group eNOS and nNOS immunoreactivity increased compared with the UCMS group. These findings support that UCMS decreases penile constitutive NOS expression via TNF-α, which may contribute to the development of ED. Blockage of TNF-α actions may represent an alternative therapeutic approach for ED in chronic psychological stress.

Effects of long-term treatment with fluoxetine and venlafaxine on rat isolated vas deferens.

Antidepressant therapy is considered as one of the factors leading to male infertility. In this study, the effects of long-term treatment with fluoxetine or venlafaxine were investigated on electrical field stimulation (EFS, 1-64 Hz), noradrenaline (10(-8) to 10(-4) M), serotonin (10(-8) to 10(-4) M), adenosine 5'-triphosphate [ATP (10(-8) to 10(-4) M)] and 80 mM KCl-induced contractile responses in the epididymal and prostatic portions of rat isolated vas deferens strips. Serotonin-induced contractile responses were significantly increased in the epididymal portion of the vas deferens obtained from the fluoxetine-treatment group, whereas in the prostatic portion there was no change. However, venlafaxine treatment had no effect on serotonin responses in the either portion of the vas deferens. Both fluoxetine and venlafaxine treatment significantly inhibited ATP-evoked contractions of the prostatic and epididymal portions of the rat vas deferens, but had no effect on EFS, noradrenaline- and KCl-evoked contractions of the vas deferentia in both portions. In conclusion, these results suggest that chronic treatment with fluoxetine and venlafaxine affects vas deferens motility. Purinoceptors may, at least in part, responsible for the impaired motility in chronic treatment of venlafaxine and fluoxetine.

Investigation of enhancement effects of nicotine on cholinergic neurotransmission in isolated rabbit gastric fundus: role of antioxidants.

Nicotine, which is tobacco alkaloid, still induces interests for researchers because of smokers addiction to nicotine. Nicotine having influence on the neuronal acetylcholine receptors (nAChRs) increases release of most certain neurotransmitters from the nerve endings. Also, nicotine, affecting the mitochondrial respiratory chains, contributes to the formation of reactive oxygen species. In the present study, we investigated the effects of nicotine on smooth muscles of gastric fundus on the electrical field stimulation (EFS) that induces transition contraction via stimulation nAChRs. In addition, we aimed to investigate the interaction between release of acetylcholine, induced by nicotine, and the effects of reactive oxygen species. Therefore, the effects of allopurinol (10(-6)-10(-5) M), deferoxamine (10(-4) M) and mannitol (10(-4)-5 x 10(-3) M) were tested on the transient contraction induced by nicotine. In conclusion, mannitol (5 x 10(-3) M) significantly reduced contractile response to nicotine on EFS only in high concentration. Whereas in small concentrations mannitol (10(-4) M) statistically did not cause any results. Deferoxamine and allopurinol also did not have any significant response.

Effects of diabetes and elevated glucose on nitrergic relaxations in the isolated duodenum of the rat.

Nitrergic relaxations of the isolated duodenum, induced by streptozotocin, were investigated in the experimental 8-week diabetes rat model. The effects of elevated glucose were also examined in the incubated duodenal muscles (in Krebs-Henseleit solution containing 44 mM glucose for 6 h) taken from nondiabetic rats. The relaxations induced by electrical field stimulation (EFS) and nicotine were significantly reduced in diabetic rats compared with control rats. Incubating of duodenal tissues in medium containing elevated glucose revealed significantly impaired relaxations to EFS and nicotine compared to responses obtained after normal glucose incubation. However, the relaxant responses to sodium nitroprusside and papaverine were similar in all groups. Incubating in hyperosmolar solutions containing sucrose, the relaxant responses were not affected. In conclusion, impairment of NO-mediated relaxations in diabetes may be related to hyperglycemia. The alterations caused by elevated glucose are not due to a hyperosmotic effect because the same concentration of sucrose had no effect on the relaxations.

Secondhand tobacco smoke impairs neurogenic and endothelium-dependent relaxation of rabbit corpus cavernosum smooth muscle: improvement with chronic oral administration of L-arginine.

The first goal of this study was to examine the effect of secondhand smoking on neurogenic, endothelium- and cGMP-dependent relaxant responses of rabbit corpus cavernosum smooth muscle. Our second goal was to determine whether such an effect can be prevented by oral administration of L-arginine. Male New Zealand rabbits were divided into control, chronic passive cigarette smoking and L-arginine treatment groups. Relaxant or contractile responses in isolated corpus cavernosum smooth muscle strips were determined by using in vitro muscle technique. There was no significant difference in the relaxant response of the strips to papaverine, sodium nitroprusside and contractile response to KCl among the groups. Relaxant responses to acetylcholine and electrical field stimulation and contractile response to phenylephrine were significantly decreased in the strips of the smoking group than that of the control group. The impaired relaxations of strips were markedly improved by treatment of L-arginine, but the contractile responses to phenylephrine were not affected. These data indicate that secondhand smoking may impair both neurogenic and endothelium-dependent relaxation of corpus cavernosum smooth muscle, and may contribute to the etiology of impotence. Chronic dietary supplementation with L-arginine offsets the impairment of neurogenic and endothelial relaxation. Therefore, we suggest that secondhand smoking exposure to cigarette produces selective impairment of neurogenic and endothelium-dependent relaxation of corpus cavernosum smooth muscle via a mechanism related to the decreased production and/or availability of nitric oxide.

Impaired esophageal reactivity in adriamycin-induced rat esophageal atresia: an in vitro study.

PURPOSE: The aim of this study was to investigate the reactivity of lower esophageal smooth muscle in the Adriamycin-induced esophageal atresia (EA) rat model. METHODS: The fetuses were divided into 3 groups. The control group was exposed to saline. The second group comprised fetuses that were exposed to Adriamycin but in whom EA did not develop. The third group comprised of fetuses that were exposed to Adriamycin and EA was observed. The reactivity of distal esophageal strips was studied in organ chambers. RESULTS: The tension was similar in all groups precontracted with carbachol for the study of relaxation to serotonin. Relaxation of lower esophageal strips to serotonin was comparably unaffected in the control and Adriamycin-no EA groups, whereas it was significantly inhibited in the EA group with decreased E(max) and pD(2) values. Contractile responses of esophageal smooth muscle to carbachol or 80 mmol/L KCl and relaxant responses to papaverine were similar in all groups. No change in agonist potency was observed among the groups. CONCLUSIONS: Our study showed impairment of serotonin-receptor-mediated relaxation; but not of cholinoceptor-mediated contraction of the lower esophageal smooth muscle in the EA. Thus, impaired relaxant responses may be, at least in part, a contributing factor in the esophageal dismotility seen in EA.

Chronic ethanol consumption impairs adrenoceptor- and purinoceptor-mediated relaxations of isolated rat detrusor smooth muscle.

OBJECTIVE: To investigate the effects of chronic ethanol consumption on the reactivity of detrusor smooth muscle. MATERIALS AND METHODS: Eight male rats received ethanol (7.2% v/v) in a modified liquid diet for 4 weeks. Two control groups were assessed; eight rats in one group were fed sucrose and received a liquid diet, and 12 rats in the second group received standard rat chow and water for 4 weeks. The reactivity of detrusor smooth muscle strips from ethanol-fed animals and control animals was evaluated in organ chambers. RESULTS: The relaxation response elicited by isoprenaline or adenosine was unaffected in the both control groups while it was significantly inhibited, with decreased maximum responses and pD2 values, in the ethanol-fed group. Contractile responses of detrusor smooth muscle to carbachol or 80 mmol/L KCl and relaxant responses to papaverine were similar in the control groups and the ethanol-fed group. There was no change in agonist potency among the groups. CONCLUSION: Chronic ethanol consumption impairs beta-adrenoceptor- and purinoceptor-mediated relaxation but not cholinoceptor-mediated contraction of the rat detrusor smooth muscle. Thus, it appears that different regulatory mechanisms are involved in ethanol-induced alterations in beta-adrenergic, purinergic and muscarinic receptors in detrusor strip.

Effects of experimental obstructive jaundice on contractile responses of dog isolated blood vessels: role of endothelium and duration of bile duct ligation.

1. We examined the effects of experimental obstructive jaundice caused by bile duct ligation (BDL) on vascular smooth muscle function, as well as the underlying mechanisms involved, by recording responses to noradrenaline (NA), 5-hydroxytryptamine (5-HT) and acetylcholine (ACh) in canine isolated renal arteries and to NA in isolated mesenteric arteries in vitro. All studies were performed 7 days after the onset of BDL in renal arteries and 3, 7 and 15 days after the onset of BDL in mesenteric arteries. 2. The maximum contraction evoked by both NA and 5-HT was significantly attenuated with no change in agonist potency (pD2 value) in renal arteries with endothelium obtained from 7 day BDL dogs when compared with those from sham-operated controls (SO). However, the reduction almost disappeared when the endothelium was removed. In contrast, no change in the responsiveness of renal arteries to KCl could be detected at 7 day BDL. Endothelium-dependent relaxations produced by ACh were significantly increased in renal artery rings from 7 day BDL dogs, but the endothelium-independent relaxations produced by papaverine in BDL preparations were not changed when compared with SO controls. 3. At 7 and 15 days after BDL, the Emax values of the mesenteric ring of BDL dogs to NA were significantly lower than that of SO controls, whereas 3 days after surgery there was no significant difference. The pD2 values in arteries obtained from 15 day BDL animals were significantly lower than those obtained from SO control animals. However, no significant changes in pD2 values were seen 3 and 7 days after the onset of BDL. 4. In conclusion, it is suggested that enhanced production and/or release of nitric oxide, mainly of endothelial origin, is associated with reduced vascular responses to contractile agents in experimental obstructive jaundice and that this effect is related to the duration of obstructive jaundice. These results may explain, at least in part, a cause of hypotension that leads to renal failure in patients with obstructive jaundice.

Effects of chronic unilateral internal pudendal arterial occlusion on reactivity of isolated corpus cavernosum strips from rabbits.

An animal model was developed to elucidate the effect of chronic obstruction of the internal pudendal artery on the responsiveness of the corpus cavernosum. In male albino rabbits, the internal pudendal artery was chronically ligated unilaterally with a silk tie and the occlusion was maintained for 1 month. The control group was sham-operated. The reactivity of corpus cavernosum tissue from the ligated animals and the control animals was studied in organ chambers. Unilateral chronic ligation of the internal pudendal artery caused an impaired contractile response to alpha-adrenoceptor stimulation with decreased Em and pD2 values and an impaired relaxant response to electrical field stimulation but resulted in a marked increase in the endothelium-dependent relaxant response to carbachol with an increased pD2 value. However chronic obstruction of the pudendal artery had no effect on adenosine-, papaverine- and sodium nitroprusside-induced relaxant responses, and there was no change in agonist potency. These data indicate that altered penile hemodynamics have an effect on the reactivity of the corpus cavernosum and may contribute to the etiology of impotence.

Impaired contraction and relaxation in the aorta of streptozotocin-diabetic rats.

It is known that diabetes mellitus alters the vascular responsiveness to several vasoconstrictors and vasodilators. Endothelium-derived nitric oxide is a potent endogenous nitrovasodilator, and endothelin-1 (ET-1) is a potent endothelium-derived vasoconstrictor substance. They play a major role in the modulation of vascular tone. Selective impairment of endothelium-dependent relaxation and impaired vasoconstriction in response to ET-1 could result in vascular disorders. The purpose of our study was to determine whether vascular responses to ET-1 and endothelium-dependent relaxing substances are impaired in rats with streptozotocin-induced diabetes of 2 weeks duration. Endothelium-dependent relaxations produced by carbachol and ATP in aortic rings precontracted with phenylephrine were significantly attenuated in rings from diabetic rats, but the endothelium-independent relaxations produced by sodium nitroprusside and adenosine in diabetic preparations were not changed when compared to the corresponding controls. The ET-1-induced contractions were significantly attenuated with no change in agonist potency (pD2 value) in aortae with and without endothelium obtained from diabetic rats when compared to those from controls. Mechanical removal of the endothelium did not significantly change ET-1 responses of aortae from either diabetic or control rats compared with responses of aortae with intact endothelium. These results suggest that, in this diabetic model, the contractile responsiveness of thoracic aortic muscles and the endothelial functions are significantly altered during 2 weeks of diabetes.

Effects of castration on adrenergic, cholinergic and nonadrenergic, noncholinergic responses of isolated corpus cavernosum from rabbit.

OBJECTIVE: To investigate the effects of castration and testosterone on the constricting effect of phenylephrine and endothelium-dependent and -independent relaxing effects of different agonists in the corpus cavernosum of male rabbits. MATERIALS AND METHODS: Twenty rabbits were castrated and 10 received testosterone replacement for 1 month after castration; 10 further rabbits underwent a sham operation and acted as controls. One month after operation the rabbits were killed and their penises excised. Strips of corpus cavernosum were used for isometric tension measurements in organ chambers; concentration-response relationships for phenylephrine, carbachol, adenosine and sodium nitroprusside were obtained by adding the reagent cumulatively to the bath. RESULTS: The phenylephrine-induced contractions were markedly lower, with no change in the pD2 values (i.e. the negative logarithm of the concentration for half-maximal response), in cavernosal strips obtained from castrated rabbits than in those from controls. Endothelium-dependent relaxation elicited by carbachol increased in the castrated group but the relaxation induced by sodium nitroprusside did not change and those elicited by adenosine were strongly depressed when compared with controls. There were no significant changes in the pD2 values of agonist-induced relaxation responses in all groups. The relaxation elicited by electrical-field stimulation at lower frequencies increased in strips from castrated rabbits but at higher frequencies were unchanged when compared with controls. Castration-induced changes in the relaxation response of cavernosal strips were significantly restored by in vivo testosterone replacement but those induced by phenylephrine were not. CONCLUSION: The lack of testosterone has an effect on the reactivity of the corpus cavernosum, indicating that testosterone has an important role in erectile function by a pre- or post-synaptic action on the corpus cavernosum.

Altered endothelium-mediated relaxation by sympathectomy in isolated rabbit carotid artery rings.

The influence of cervical and periarterial sympathectomy on endothelium-dependent and endothelium-independent relaxations of the mature rabbit carotid artery was studied in vitro. The responses to adenosine, vasoactive intestinal polypeptide and substance P in sympathectomized and control rabbit carotid artery rings were recorded and analyzed. The effects of endothelium removal were also investigated. The maximal relaxation achieved by substance P, which produces endothelium-dependent relaxation, was significantly inhibited in 3 weeks in postsympathectomy arterial preparations as compared to controls. Adenosine and vasoactive intestinal polypeptide, which produce endothelium-independent relaxation, elicited similar relaxation in all tissues. These results demonstrated that the response to substance P was impaired by cervical and periarterial sympathectomy. The decreased maximum response to substance P may be the result of a decreased NK-1 receptor subtype density or excitation/response coupling, or it may be due to an impaired production and/or liberation of endothelium-derived relaxing factor (EDRF).