Brain nitric oxide production by a proline-rich decapeptide from Bothrops jararaca venom improves baroreflex sensitivity of spontaneously hypertensive rats.

Baroreflex sensitivity is disturbed in many people with cardiovascular diseases such as hypertension. Brain deficiency of nitric oxide (NO), which is synthesized by NO synthase (NOS) in the citrulline-NO cycle (with argininosuccinate synthase (ASS) activity being the rate-limiting step), contributes to impaired baroreflex. We recently showed that a decapeptide isolated from Bothrops jararaca snake venom, denoted Bj-PRO-10c, exerts powerful and sustained antihypertensive activity. Bj-PRO-10c promoted vasodilatation dependent on the positive modulation of ASS activity and NO production in the endothelium, and also acted on the central nervous system, inducing the release of GABA and glutamate, two important neurotransmitters in the regulation of autonomic systems. We evaluated baroreflex function using the regression line obtained by the best-fit points of measured heart rate (HR) and mean arterial pressure (MAP) data from spontaneously hypertensive rats (SHRs) treated with Bj-PRO-10c. We also investigated molecular mechanisms involved in this effect, both in vitro and in vivo. Bj-PRO-10c mediated an increase in baroreflex sensitivity and a decrease in MAP and HR. The effects exerted by the peptide include an increase in the gene expression of endothelial NOS and ASS. Bj-PRO-10c-induced NO production depended on intracellular calcium fluxes and the activation of a G(i/o)-protein-coupled metabotropic receptor. Bj-PRO-10c induced NO production and the gene expression of ASS and endothelial NOS in the brains of SHRs, thereby improving baroreflex sensitivity. Bj-PRO-10c may reveal novel approaches for treating diseases with impaired baroreflex function.

The central nervous system as target for antihypertensive actions of a proline-rich peptide from Bothrops jararaca venom.

Pyroglutamyl proline-rich oligopeptides, present in the venom of the pit viper Bothrops jararaca (Bj-PROs), are the first described naturally occurring inhibitors of the angiotensin I-converting enzyme (ACE). The inhibition of ACE by the decapeptide Bj-PRO-10c (

Platelet-derived exosomes from septic shock patients induce myocardial dysfunction.

INTRODUCTION: Mechanisms underlying inotropic failure in septic shock are incompletely understood. We previously identified the presence of exosomes in the plasma of septic shock patients. These exosomes are released mainly by platelets, produce superoxide, and induce apoptosis in vascular cells by a redox-dependent pathway. We hypothesized that circulating platelet-derived exosomes could contribute to inotropic dysfunction of sepsis. METHODS: We collected blood samples from 55 patients with septic shock and 12 healthy volunteers for exosome separation. Exosomes from septic patients and healthy individuals were investigated concerning their myocardial depressant effect in isolated heart and papillary muscle preparations. RESULTS: Exosomes from the plasma of septic patients significantly decreased positive and negative derivatives of left ventricular pressure in isolated rabbit hearts or developed tension and its first positive derivative in papillary muscles. Exosomes from healthy individuals decreased these variables non-significantly. In hearts from rabbits previously exposed to endotoxin, septic exosomes decreased positive and negative derivatives of ventricular pressure. This negative inotropic effect was fully reversible upon withdrawal of exosomes. Nitric oxide (NO) production from exosomes derived from septic shock patients was demonstrated by fluorescence. Also, there was an increase in myocardial nitrate content after exposure to septic exosomes. CONCLUSION: Circulating platelet-derived exosomes from septic patients induced myocardial dysfunction in isolated heart and papillary muscle preparations, a phenomenon enhanced by previous in vivo exposure to lipopolysaccharide. The generation of NO by septic exosomes and the increased myocardial nitrate content after incubation with exosomes from septic patients suggest an NO-dependent mechanism that may contribute to myocardial dysfunction of sepsis.

The herbal drug Catuama reverts and prevents ventricular fibrillation in the isolated rabbit heart.

INTRODUCTION: Catuama, an herbal drug very popular in Brazil, was tested on the reversion and prevention of ventricular fibrillation (VF) in the isolated rabbit heart. MATERIALS AND METHODS: Catuama (a mixture of Trichilia catigua, Paullinia cupana, Ptychopetalum olacoides, and Zinziber officinalis) was perfused in the isolated perfused rabbit heart. Its effects on intraventricular conduction, heart rate, and monophasic action potential (MAP) duration were evaluated, and sustained VF was induced. The effects on reversion and reinduction of arrhythmia were observed, and new measures were taken in the hearts that reverted. RESULTS: Catuama and T catigua reverted VF in all hearts, prevented reinduction, and prolonged intraventricular conduction. Catuama prolonged MAP phase 2. On the other hand, P cupana reverted VF in 3 of 5 hearts, but depressed automatism, prolonged MAP phase 3, and did not prevent reinduction. DISCUSSION: Catuama reverted and prevented VF in this model. T catigua extract is probably the main agent responsible for the beneficial actions observed. Further studies are now in progress to clarify these actions.

Hypertonic saline solution increases the expression of heat shock protein 70 and improves lung inflammation early after reperfusion in a rodent model of controlled hemorrhage.

Hypertonic saline solution (HS solution, NaCl 7.5%) has shown to restore hemodynamic parameters in hemorrhagic shock and to decrease the inflammation triggered by ischemia-reperfusion injury (I-R). Therefore, our objective was to investigate the effects of HS solution on the mechanisms involved in I-R, in an experimental model of controlled hemorrhagic shock. Wistar rats (280-350 g) were submitted to controlled bleeding, keeping the mean arterial pressure around 40 mmHg, for 1 h. After that, rats were randomized and treated with HS solution (4 mL/kg) or normal saline (34 mL/kg). There were no differences in hemodynamic parameters between both groups for at least 2 h after shock. No difference either was observed in reactive oxygen species generation (measured indirectly by malondialdehyde concentration) or cytokines (interleukins 6 and 10) production (measured by enzyme-linked immunosorbent assay). Quantitative analysis of lung tissue showed a smaller neutrophil infiltration in animals that received HS solution. Moreover, the animals in the HS group showed an increased expression of heat shock protein 70. Therefore, we concluded that treatment of hemorrhagic shock with HS solution can decrease pulmonary inflammation and increase cellular protection by up-regulating heat shock protein 70 expression.

Local and systemic effects of hypertonic solution (NaCl 7.5%) in experimental acute pancreatitis.

OBJECTIVES: Severe acute pancreatitis (AP) is characterized by hemodynamic alterations and a systemic inflammatory response, leading to a high mortality rate. Treatment of hemorrhagic shock with hypertonic saline solutions significantly reduces mortality through an improvement in the hemodynamic conditions and possibly by an anti-inflammatory effect. Therefore, hypertonic solutions could be effective in AP. METHODS: Wistar rats were divided in 4 groups: group C, control, without AP; group NT, AP, without treatment; group NS, treatment with normal saline solution (NaCl 0.9%) 1 hour after AP; group HTS, treatment with hypertonic saline solution (NaCl 7.5%) 1 hour after AP. AP was induced by injection of 2.5% sodium taurocholate into the pancreatic duct. Mean arterial blood pressure (MAP) and heart rate were recorded at 0 and 2, 4, 24, and 48 hours after AP. After induction of AP, animals were killed at 2, 12, 24, and 48 hours for serum amylase, interleukin (IL)-6, and IL-10 analysis, pancreatic tissue culture and histologic analysis, oxidation and phosphorylation of liver mitochondria, pulmonary myeloperoxidase activity (MPO), and mortality study. RESULTS: In animals of groups NS and NT, a significant decrease of MAP was observed 48 hours after AP (NS: 91 +/- 3 mm Hg; NT: 89 +/- 3 mm Hg) compared with baseline (C: 105 +/- 2 mm Hg) and to HTS group (HTS: 102 +/- 2 mm Hg; P < 0.05). In animals of group NT, NS, and HTS, serum IL-6 and IL-10 levels were significantly higher at 2 hours after AP compared with the control group. However, IL-6 levels at 12 hours after AP and IL-10 levels at 2 and 12 hours after AP were significant lower in group HTS compared with NS and NT groups (P < 0.05). In group HTS, a decrease of pulmonary MPO activity and of pancreatic infection was observed 24 hours after AP compared with NT and NS groups (P < 0.05). A significant reduction on pancreatic acinar necrosis and mitochondrial dysfunction was observed after 48 hours of AP in animals of group HTS compared with groups NT and NS (P < 0.05). A significant reduction on mortality was observed in HTS (0/14) compared with NS (6/17; 35%) and NT (7/20; 35%). CONCLUSIONS: The administration of hypertonic saline solution in experimental AP attenuated hemodynamic alterations, decreased inflammatory cytokines, diminished systemic lesions and pancreatic acinar necrosis, prevented pancreatic infection, and reduced the mortality rate.

Efeito da solução hipertônica de NaCI 7, 5 por cento na resposta inflamatória em modelo de choque hipovolêmico / Effects of hypertonic solution (NaCI 7, 5 per cent) on the inflammatory response in an experimental model of hemorrhagic shock

A solução hipertônica de cloreto de sódio 7,5 por cento (SSH) é eficaz em restaurar os parâmetros hemodinâmicos e reduzir a inflamação em modelos experimentais de choque hemorrágico. Assim, foi nosso objetivo investigar a ação da SSH sobre os mecanismos envolvidos na lesão de isquemia e reperfusão (I/R) em um modelo de choque hemorrágico controlado. Ratos Wistar (280-350 g) foram submetidos à hemorragia controlada, mantendo-se a pressão arterial média em 40 mmHg por 1 h / Hypertonic saline solution (HSS - NaCI 7,5 per cent) was shown to restore hemodynamic parameters in hemorrhagic shock and to decrease the inflammation triggered by ischemia-reperfusion injury (I/R). Therefore, our objective was to investigate the effects of HSS on the mechanisms involved in I/R, in an experimental model of controled hemorrhagic shock. Wistar rats (2`80-350 g) were submitted to the controled bleeding, keeping the mean arterial pressure around 40 mmHg, for 1 hour...

New model of ventricular fibrillation.

The purpose of this study was to develop a more efficient and stable model of ventricular fibrillation (VF) in the isolated rabbit heart, because there is not a satisfactory model with this animal. We also observed the effects of increasing extracellular calcium in the stability and reversibility of the arrhythmia. After suspending the hearts in a classical Langendorff preparation, VF was induced by burst stimulation (current = 2.0 mA, pulse duration = 3 milliseconds, frequency = 50 Hz, voltage = 10 V, duration of stimulation = 5 minutes). The hearts were then divided into 2 groups, A and B. The hearts in group B were perfused with a modified Krebs-Henseleit solution, which contained twice as much calcium as the solution used in the other group. The rate of success with this model was 100% for both groups. The hearts fibrillated up to 30 minutes in group A and more than 40 minutes in group B, longer then all studies ever published in rabbit hearts. Ventricular fibrillation reverted to sinus rhythm in 100% of the hearts of group A when treated with an antifibrillatory drug, whereas no reversion at all was observed in the hearts of group B. We conclude that high extracellular calcium makes the reversion to sinus rhythm more difficult in this model. Our high rate of success and the exceptionally stable and long-lasting VF turn our model very effective for the study of antiarrhythmic interventions in the isolated rabbit heart.