Marcos R. Ferraz

Depression and Cardiovascular Disease: Role of Nitric Oxide

Both depression and cardiovascular disease are major public health problems. Growing evidence shows that depression is a risk factor for the development of coronary artery disease (CAD). However, the exact mechanisms underlying the interplay between depression and CAD remain to be elucidated. Depression adversely affects autonomic and hormonal homeostasis, resulting in metabolic abnormalities, inflammation, increased platelet aggregation and endothelial dysfunction. All of these pathological features lead to atherothrombosis and cardiovascular events. However, there is no clear evidence that anti-depressant drugs or psychotherapy will reduce the risk or improve the outcome of CAD. Recent studies suggest that the L-arginine-nitric oxide (NO) pathway is involved in the genesis of depression. NO has many physiological functions, including vasodilatation, neurotransmission and platelet aggregation inhibition. It is synthesised from the cationic amino acid L-arginine by a family of enzymes: NO synthases (NOS). There are three NOS isoforms: inducible NOS (iNOS), endothelial NOS and neuronal NOS (nNOS). The availability and transport of L-arginine modulate rates of NO biosynthesis in circulating blood cells and vasculature, which provides a protective effect against cardiovascular disease. In depressive patients, the L-arginine-nitric oxide pathway seems to be impaired. The present review seeks a better understanding of the mechanisms that could identify depression as a cardiovascular risk factor and introduce new possible therapeutic interventions.

How REM sleep deprivation and amantadine affects male rat sexual behavior.

There are conflicting findings about the sexual effects of REM sleep deprivation (REMd). Otherwise, several studies show a dopaminergic hypersensitivity after REMd. The effect of REMd and amantadine (AMA) was studied for standard measures and temporal patterning in the first experiment, in four groups: normal with vehicle, normal with AMA (5.0 and 10 mg/kg), REMd with vehicle and REMd with AMA (5.0 and 10.0 mg/kg). REMd reduced mount latency (ML), intromission latency (IL) and mount number (MN) and increased copulatory efficiency (CE) and hit rate factor. REMd also reduced the mount bout number (MBN) and increased the sexual interaction (mount bout time, MBT) among male and female during copula. AMA stimulates initiation and hit rate factors and accelerates the temporal patterning of sexual behavior, evoking fewer and quicker mount bouts. In the experiments with combination of REMd and AMA administration, AMA did not increase behavior effects evoked by REM deprivation, probably due to a top or a bottom effect, depending on the measures considered. A second experiment studied the effects of AMA (1.25 to 5.0 mg/kg) and REMd on the sexual reflexes of nonimmobilized male rats. REMd enhanced the AMA effects upon the seminal emission reflex, but inhibited the penile erection reflex elicited by 1.25 mg/kg of AMA. Curiously, our results showed that REMd, like AMA, a dopaminergic agonist, causes similar effects of sexual behavior in the male rat, particularly those related to arousal mechanism and hit rate factor. The results are discussed and the effects of REMd probably involve dopaminergic hypersensitivity and increased sexual motivational response.

Low plasma levels of l-arginine, impaired intraplatelet nitric oxide and platelet hyperaggregability: Implications for cardiovascular disease in depressive patients

BACKGROUND Major depression (MD) is an independent cardiovascular risk factor, but the exact mechanisms are not clear. In this study we have investigated the intraplatelet L-arginine-nitric oxide (NO) pathway and platelet function in depressive patients. METHODS Nineteen unmedicated patients with MD (34±4years) and 19 control subjects (CS, 34±3years) were included. L-[(3)H]-arginine influx, NO synthase (NOS) activity and intracellular cGMP levels were evaluated in platelets, as well as the expression of eNOS, iNOS, arginase and soluble guanylate cyclase (sGC), platelet aggregation and the systemic amino acid profile in MD patients and CS. RESULTS L-arginine influx (pmol/10(9)cells/min) in platelets was reduced from 46.2±9.5 to 20.02±2.12 in depression. NOS activity (pmol/10(8) cells) was diminished in MD patients (0.09±0.01) compared to CS (0.17±0.01). Intracellular cGMP levels were also impaired in MD patients associated with hyperaggregability. Moreover, the concentration of plasma L-arginine was reduced by 20% in MD patients. The expression of eNOS, iNOS, arginase II and sGC in platelet lysates was not affected by MD. LIMITATIONS Small number of patients in the study. CONCLUSIONS This study has demonstrated an impairment of L-arginine-NO signaling in platelets from MD patients, suggesting a role in platelet activation and cardiovascular events.

Amantadine stimulates sexual behavior in male rats

The effects of amantadine on sexual behavior, penile erection, and seminal emission of male rats was studied. Amantadine significantly decreased latency of mounts in all doses (1.25 to 50 mg/kg), and decreased the number of mounts and intromission latency at the highest doses used. The lowest dose of amantadine significantly increased ejaculation latency and intromission frequency, while higher doses significantly reduced it, which indicates a biphasic response of the drug. Additionally, seminal emission, erections, and genital grooming were significantly induced by amantadine. Amantadine-induced seminal emissions were impaired by spinal cord transection, which suggests the involvement of supraspinal structures in the drug action. Haloperidol and atropine sulphate significantly reduced seminal emissions and penile erections induced by amantadine. These results demonstrate that amantadine stimulates sexual behavior and genital reflexes in male rats and suggest a facilitatory effect of the drug that probably involves different mechanisms of action.

Major depression induces oxidative stress and platelet hyperaggregability.

We have previously demonstrated an impairment of intraplatelet L-arginine-nitric oxide-cGMP pathway in major depression (MD) associated to platelet dysfunction. Here, we evaluated arginase pathway and phosphodiesterase 5 (PDE5) expression in platelets, systemic and intraplatelet oxidative status in untreated MD patients, and their effects on platelet aggregation. Blood samples were collected from 22 treatment naive MD patients (31 ± 2 yr) and 27 healthy subjects (33 ± 2 yr). MD patients presented with an activation of platelet arginase II, which competes with L-arginine for the production of nitric oxide (NO). An increase in protein carbonylation, overexpression of NADPH oxidase and PDE5, an enzyme that inactivates cGMP, was observed in platelets from MD patients compared to controls. In this context, platelet hyperaggregability was found in MD patients. On the other hand, antioxidant enzymes catalase, glutathione peroxidase and superoxide dismutase activities in serum and in platelets did not differ between groups. The increased activation of intraplatelet arginase and platelet aggregability, in addition to an overexpression of PDE5 and oxidative stress may contribute to alterations in L-arginine-NO-cGMP pathway and in platelet function, and consequently to the increased thrombotic risk in MD.

Defective nitric oxide-cyclic guanosine monophosphate signaling in patients with bipolar disorder: a potential role for platelet dysfunction.

Objective Bipolar disorder (BD) is associated with elevated cardiovascular mortality rates. We investigated the modulation of L-arginine–nitric oxide (NO) signaling in platelets from patients with BD at different phases. Methods Platelets obtained from 28 patients with BD and 10 healthy volunteers were analyzed for L-arginine transport, NO synthase (NOS) activity, cyclic guanosine monophosphate content, and biomarkers of oxidative stress. Expressions of NOS isoforms, soluble guanylyl cyclase, and arginase were also measured in platelets. Amino acid and C-reactive protein levels in plasma were assessed. Results Plasma concentrations of L-arginine (mean [M] ± standard error of the mean [SEM] = 97 ± 10 versus 121 ± 10 µM) and its transport into platelets (median [interquartile range] = 26.0 [28.6] versus 26.5 [43.9] pmol/109 cells per minute) did not differ between patients with BD and controls (p > .05). Patients with BD showed reduced NOS activity (M ± SEM = 0.037 ± 0.003 versus 0.135 ± 0.022 pmol/108 cells, p < .001), but not endothelial NOS, inducible NOS, and arginase expression, compared with controls (p > .05). Cyclic guanosine monophosphate content was reduced (M ± SEM = 0.022 ± 0.003 versus 0.086 ± 0.020 pmol/108 cells, p < .05) despite the absence of changes in soluble guanylyl cyclase expression (median [interquartile range] = 21.6 [15.5] versus 9.5 [9.4] arbitrary units, p > .05) in patients with BD. Superoxide dismutase activity, but not catalase activity, was increased in patients with BD in the manic phase (M ± SEM = 2094 ± 335 versus 172 ± 17 U/mg protein, p < .001). C-reactive protein was elevated only in manic episodes (M ± SEM = 0.8 ± 0.2 versus 0.1 ± 0.02 mg/L, p < .001). Conclusions Impaired NO generation from platelets, inflammation, and oxidative stress may play pivotal roles in the multifaceted process of cardiovascular events in BD. Abbreviations NO = nitric oxide NOS = nitric oxide synthase BD = bipolar disorder

Chronic amantadine treatment enhances the sexual behaviour of male rats

The acute administration of amantadine (AMA), a dopaminomimetic and NMDA glutamatergic receptor antagonist also used as an anti-Parkinsonian agent, stimulates male rat sexual behaviour. However it remains unclear whether long term AMA supplementation might also provoke a similar increase in male rat sexual conduct. In the present study, male rats were administered AMA (5-50 mg/kg/day) or vehicle daily for 21 days and their sexual response was monitored weekly. Chronic treatment with AMA effectively increased the sexual response of male rats, similarly to what had been observed before with acute amantadine treatment. The main effect of chronic AMA treatment occurs in arousal and in ejaculatory response, whilst the excitatory component was not affected. The 21-day treatment with AMA did not lead to tolerance, suggesting that perhaps AMA could be used in male human patients to prevent sexual inhibition caused by anti-depressant and anti-psychotic agents.

Preventing L-NAME inhibitory effects on rat sexual behavior with hydralazine, isradipine or captopril co-treatment.

The effects of the chronic oral treatment with N(G)-nitro-L-arginine methyl ester (L-NAME), separately or in combination with isradipine, captopril or hydralazine, on standard and temporal patterning sexual behavior of male rats were evaluated. L-Arginine and filtered water were used as control. L-NAME treatment decreased the copulatory rate and hit rate factors of sexual behavior. However, the initiation factor and temporal patterning were less modified by the drug. After 14 days of L-NAME treatment suspension the male rat sexual response was recovered. The three antihypertensive agents were able to reverse partially or totally the inhibitory effects of L-NAME, suggesting that the chronic oral treatment with L-NAME induces penile erection dysfunction by peripheral mechanisms. The present results suggest that chronic oral treatment with nitric oxide (NO) synthase inhibitor can be a relevant and powerful peripheral erectile dysfunction model to evaluate the effects of drugs on erectile function of male rats.

Prenatal hypoxia, habituation memory and oxidative stress

Hypoxia-ischemia (HI) is characterized by a reduced supply of oxygen during pregnancy, which leads to both central nervous system and peripheral injuries in the foetus, resulting in impairment in its development. The purpose of this study was to investigate behavioural changes and systemic oxidative stress in adult animals that have been affected by HI during pregnancy. HI was induced by the occlusion of the maternal uterine artery with aneurysm clamps for a period of 45 min on the 18th gestational day. Animals from the sham group were submitted to same surgical procedure as the HI animals, without occlusion of the maternal uterine artery. The control group consisted of non-manipulated healthy animals. At postnatal day 90, the pups were submitted to behavioural tests followed by blood collection. HI adult animals presented an increase in anxiety behaviour and a lack of habituation compared to both sham and control groups. Oxidative damage, assessed by protein and lipid oxidation in serum, did not differ between HI and sham-operated animals. However, HI animals presented reduced activity of the glutathione peroxidase enzyme and increased formation of nitrite, indicating alterations in the systemic antioxidant repair system. Our results suggest an association among HI, systemic oxidative stress and behavioural alterations.

Sleep deprivation affects sexual behavior and tyrosine hydroxylase (TH) levels in sexually experienced male rats.

Paradoxical sleep deprivation (PSD) produces alterations in dopaminergic systems and also modifies sexual behavior. In this work we evaluated PSD effects on the sexual response and tyrosine hydroxylase (TH) expression in dopaminergic pathways related to sexual behavior of naive and sexual experienced rats. Male Wistar rats had their sexual behavior evaluated in 6 copulatory tests, with a 4 days interval. In these tests, the animals interacted with a receptive female and parameters that compose each component of the male sexual reply (initiation, arousal and ejaculation) were evaluated. After the 5th test, the animals were randomly divided in 2 groups, control and PSD, and 96 h later they were submitted to the last copulatory test. PSD facilitated the excitatory and the ejaculatory component, increasing the copulatory efficiency. In addition, reduced mount frequency and ejaculation latency were observed. The temporal patterning of the sexual behavior was modified, with reduction in the number of mount bouts. PSD per se was not able to modify TH levels, but in PSD sexual trained rats, an increase in the number of TH-immunoreactive cellular bodies in all dopaminergic areas evaluated was detected. Our data suggest that PSD facilitates the sexual response and this facilitation combined to sexual training could be the consequence of increased TH levels in dopaminergic pathways related to sexual reply.