Simvastatin and dehydroepiandrosterone sulfate effects against hypoxic pulmonary hypertension are not additive.

Pulmonary hypertension is a group of disorders characterized by elevated mean pulmonary artery pressure (mPAP) and pulmonary vascular resistance. To test our hypothesis that combining two drugs useful in experimental pulmonary hypertension, statins and dehydroepiandrosterone sulfate (DHEA S), is more effective than either agent alone, we induced pulmonary hypertension in adult male rats by exposing them to hypoxia (10%O2) for 3 weeks. We treated them with simvastatin (60 mg/l) and DHEA S (100 mg/l) in drinking water, either alone or in combination. Both simvastatin and DHEA S reduced mPAP (froma mean±s.d. of 34.4±4.4 to 27.6±5.9 and 26.7±4.8 mmHg, respectively), yet their combination was not more effective (26.7±7.9 mmHg). Differences in the degree of oxidative stress (indicated by malondialdehydeplasma concentration),the rate of superoxide production (electron paramagnetic resonance), or blood nitric oxide levels (chemiluminescence) did not explain the lack of additivity of the effect of DHEA S and simvastatin on pulmonary hypertension. We propose that the main mechanism of both drugs on pulmonary hypertension could be their inhibitory effect on 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase, which could explain their lack of additivity.

Chronic hypoxia increases fetoplacental vascular resistance in rat placenta perfused with blood.

BACKGROUND: Chronic hypoxia elevates vascular resistance on the fetal side of the placenta. However, when a low-viscosity perfusate is used, the increase in resistance caused by chronic hypoxia is relatively small (12 mmHg). In the present study, we tested the hypothesis that perfusion with more viscous fluid (blood) will reveal more substantial effect of chronic hypoxia. METHODS: Using an isolated, dually perfused rat placenta perfused at a constant flow rate with homologous blood, perfusion pressure on the fetal side was measured. Then, the relationship between perfusion pressure and flow (P/Q) was determined. RESULTS: Fetoplacental vascular resistance was increased by chronic hypoxia (10 % O2) during the last third of gestation. This was observed when the placentas were perfused with a low viscosity Krebs solution and more enhanced when perfused with blood. Nevertheless, we found no clear advantage for using blood instead of Krebs solution to study the effects of hypoxia on the fetoplacental vasculature. The elevation of fetoplacental vascular resistance caused by chronic hypoxia was at least partly resistant to acute reoxygenation. CONCLUSION: Using blood for the perfusion of the isolated rat placenta does not confer any clear methodological advantage over using Krebs solution (Tab. 2, Fig. 2, Ref. 21).

Depletion of alveolar macrophages attenuates hypoxic pulmonary hypertension but not hypoxia-induced increase in serum concentration of MCP-1.

Exposure to hypoxia, leading to hypoxic pulmonary hypertension (HPH), is associated with activation of alveolar macrophages (AM). However, it remains unclear how AM participate in this process. There are studies which imply that the AM product monocyte chemoattractant protein-1 (MCP-1) plays an important role. Thus we tested: 1. if the selective elimination of AM attenuates HPH in rats, 2. the correlation of MCP-1 plasmatic concentrations with the presence and absence of AM during exposure to hypoxia, 3. the direct influence of hypoxia on MCP-1 production in isolated AM. We found that experimental depletion of AM attenuated the chronic hypoxia-induced increase in mean pulmonary arterial pressure, but did not affect the serum MCP-1 concentrations. Furthermore, the MCP-1 production by AM in vitro was unaffected by hypoxia. Thus we conclude that AM play a significant role in the mechanism of HPH, but MCP-1 release from these cells is most likely not involved in this process. The increase of MCP-1 accompanying the development of HPH probably originates from other sources than AM.

L-arginine in combination with sildenafil potentiates the attenuation of hypoxic pulmonary hypertension in rats.

Chronic hypoxia induces an increased production of nitric oxide (NO) in pulmonary prealveolar arterioles. Bioavailability of the NO in the pulmonary vessels correlates with concentration of L-arginine as well as activity of phosphodiesterase-5 enzyme (PDE-5). We tested a hypothesis whether a combination of L-arginine and PDE-5 inhibitor sildenafil has an additive effect in reduction of the hypoxic pulmonary hypertension (HPH) in rats. Animals were exposed to chronic normobaric hypoxia for 3 weeks. In the AH group, rats were administered L-arginine during chronic hypoxic exposure. In the SH group, rats were administered sildenafil during chronic hypoxic exposure. In the SAH group, rats were treated by the combination of L-arginine as well as sildenafil during exposure to chronic hypoxia. Mean PAP, structural remodeling of peripheral pulmonary arterioles (%DL) and RV/LV+S ratio was significantly decreased in the SAH group compared to hypoxic controls even decreased compared to the AH and the SH groups in first two measured parameters. Plasmatic concentration of cGMP and NOx were significantly lower in the SAH group compared to hypoxic controls. We demonstrate that NO synthase substrate L-arginine and phosphodiesterase-5 inhibitor sildenafil administered in combination are more potent in attenuation of the HPH compared to a treatment by substances given alone.

Rho-kinase inhibition attenuates acute hypoxic fetoplacental vasoconstriction in the rat.

The vessels on the fetal side of the placenta differ from most other vascular beds except the lungs in that they respond to acute hypoxia by vasoconstriction. An essential role of calcium influx in the mechanism of this hypoxic fetoplacental vasoconstriction (HFPV) has been shown previously. That finding does not, however, exclude the possible involvement of other mechanisms of vascular tone regulation. In this study we tested the hypothesis that Rho-kinase-mediated calcium sensitization is involved in HFPV. We used a model of isolated rat placenta dually perfused (from both the maternal and fetal side) with Krebs salt solution saturated with normoxic and hypoxic gas mixture respectively at constant flow rate. Rho-kinase pathway was inhibited by fasudil (10 microM). We found that fasudil reduced basal normoxic fetoplacental vascular resistance and completely prevented HFPV. This suggests that the activity of Rho-kinase signaling pathway is essential for HFPV.

[Traumatic brain injuries--effects of alcohol and caffeine on intracranial pressure and cerebral blood flow]. / Traumatická poranení mozku--vliv alkoholu a kofeinu na intrakraniální tlak a prutok krve mozkem.

INTRODUCTION: Alcohol and caffeine is intermittently or regularly used by majority of people. These drugs may have acute or chronic effects on patients with traumatic brain injury. Alcohol intoxication increases cerebral blood flow from 8 to 24%. Caffeine decreases cerebral blood flow from 10 to 20%. These facts create a theoretical hypothesis that the decrease of CBF may reduce incranial pressure. The aim of this study was to investigate the effect of caffeine on intracranial pressure in rats following traumatic brain injury. METHODS: Ten Wistar male rats underwent weight drop closed head injury. The second stage of the experiment was done 12 to 18 hours later. Intracranial pressure, respiration, heart rate and the mean arterial pressure was monitored. Intraperitoneal injection of caffeine (20 mg/kg) was administered and repetitive measurement of vital signs and intracranial pressure was done. RESULTS: The baseline ICP after head injury was 8.5 +/- 2.9 mm Hg. Ten minutes after intraperitoneal caffeine administration ICP dropped to 7.6 +/- 3.1 mm Hg (p < 0.05). This represents a 11% decrease from baseline value. Mean arterial pressure, respiration and heart rate were stable. CONCLUSION: Intracranial pressure decrease of 11% from baseline value. This drop was compatible with our hypothesis. The exact dosage of caffeine, its form and rate, should be more precisely specified in further studies.

Level of vigilance influences licking frequency in rats.

In rats, the basic licking rhythm is generated by the central pattern generator located in the brainstem. Nevertheless, the licking frequency can be regulated between about 7.5 and 4 Hz by changing the drinking conditions. If these conditions are kept constant, the licking frequency can be influenced only to a minor degree by factors such as deprivation level, type of solution, and phase of the session. The aim of our study was to compare the licking frequency of rats at different levels of vigilance. We investigated spontaneous licking of rats by an electrical lick sensor; parallel behavior monitoring was also performed. Animals kept in a stable environment and showing a lower level of vigilance licked at a rate of 5.96 Hz, fully vigilant rats licked significantly more rapidly at a frequency 6.57 Hz. The fastest rate of licking (6.49 Hz and 6.82 Hz, respectively) was encountered in alert rats under a mild stress caused by the presence of a second animal in the experimental box. The vigilance level is thus another factor affecting the licking rate of rats that should be taken into account in behavioral licking experiments.

The effect of direct administration of drugs into the licking generator in rats.

Spontaneous licking of thirsty rats was investigated under the effect of GYKI (an AMPA/kainate receptor antagonist) and of 2-amino-5-phosphonopentanoic acid (AP5, an NMDA receptor antagonist) applied intracranially into the central rhythm generator of licking. Adult Long Evans male rats were stereotaxically implanted with guiding cannulae aimed at the oral part of nucleus reticularis gigantocellularis (NRG). After a recovery (1 week at least), animals after 24 h water deprivation were trained to drink in an experimental box. Then 1 microl of GYKI (1 mM solution) or 1 microl of AP5 (20 mM solution) was administrated by microinjection through the guiding cannula directly into the target structure. Lick-lick interval (LLI) was recorded by an electrical lick sensor and analysed with a laboratory computer Pentium. Localisation of the administration was checked by a routine histological method. GYKI administration significantly prolonged the LLI i.e. slowed down licking frequency. The effect was immediate and began to dwindle in the period between 10 min to 2 h. Licking frequency under the influence of AP5 was faster (shorter LLI). This effect culminated after 30 min and almost disappeared after 2 h. Both our findings are in a good accordance with those of Grillner's group that NMDA receptors are important for slow swimming movements while non-NMDA receptors are responsible for fast ones.

Functional ablation of deep cerebellar nuclei temporarily impairs learned coordination of forepaw and tongue movements.

The role of the cerebellum in complex skilled movements was assessed by the use of functional ablation technique. Rats were trained to synchronize tongue and forepaw movements in a drinking box equipped with a retractable spout which was automatically withdrawn after every lick but could be returned by pressing and releasing a lever placed 4 cm below the spout. The animals learned to perform short presses synchronized with the lick cycle in such a way as to allow continuous drinking. The contribution of the neocerebellum to these lick-associated instrumental movements was estimated by intracranial injection of 2 ng of tetrodotoxin into the dentate and lateral part of interposed nuclei. Bilateral blockade of the mainly neocerebellar output interfered with learned synchronization of licking and bar pressing, but did not suppress licking from a stationary spout and only decreased the licking frequency by 10%. It is concluded that the tongue-forepaw synchronization is disrupted by elimination of the neocerebellar output but for a much shorter time (< 9 h) than the tetrodotoxin-induced inactivation of the lateral part of the caudate nucleus (72 h) reported earlier. The results confirm participation of cerebellar hemispheres in learned tongue-forepaw synchronization, but indicate at the same time that elimination of this link can be easily compensated.