Biomedical subjects

R Przewłocki

Publications and source records attributed to R Przewłocki.

At least 19 recordsLinked to original sources

PubMed1996

Endogenous opioids may modulate the activity of the hypothalamus-pituitary-adrenocortical axis in domestic fowl.

The activity of the hypothalamus-pituitary-adrenocortical (HPA) axis is under complex neuronal, hormonal and peptidergic control. In order to determine the role of the endogenous opioids in the modulation of the HPA axis in hens we have examined the changes of the plasma levels of Met-enkephalin, alpha-neo-endorphin, catecholamines and corticosterone during rest, stress (30 min of overcrowding) and after naltrexone pretreatment. Short overcrowding induced an increase of all parameters, but the time of response and duration of elevation was different. Plasma Met-enkephalin showed a biphasic response: decrease followed by increase at the end of stress. Catecholamines and corticosterone peaked at 30 min of overcrowding and the highest level of alpha-neo-endorphin has been noticed at 20 min after the experiment was started. Naltrexone (2 mg/kg b.w., i.v) diminished the responses to stress of all parameters and did not change their basal levels, except for corticosterone. These results indicate that the endogenous opioids may take part in the mediation of HPA activity in hens during resting and stressful situations.

Animals↗

PubMed1995

Kappa opioid receptor agonists suppress absence seizures in WAG/Rij rats.

Involvement of the kappa opioid receptor in the regulation of epileptic activity was studied in WAG/Rij rats, a genetic model of absence epilepsy. I.c.v. administration of the kappa agonists U50,488H (trans-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl)-cyclohexyl]- benzeneacetamide), U69,593 (5 alpha, 7 alpha, 8 beta)-(-)-N-methyl-(1-pyrrolidinyl)-1- oxaspiro(4,5)dec-8-yl)benzeneacetamide) or PD117,302 ((+/-)-trans-N-methyl-N-[2-(1-pyrrolidinyl)- cyclohexyl]benzo[b]thiophene-4-acetamide), 50 and 150 micrograms/5 microliter each, dose-dependently decreased the number and mean duration of spike wave discharges (SWD). Peripheral administration of U50,488H (10 and 30 mg/kg s.c.) also attenuated the seizure activity in this model. The specific kappa opioid receptor antagonist nor-binaltorphimine (Nor-BNI, 10 micrograms/5 microliters i.c.v., 18 h before EEG registration) moderately increased the number of SWD, which suggests that endogenous opioids acting through kappa receptors may tonically inhibit the seizure activity in these rats. In addition, the enhancement of an absence-like seizure activity induced by the specific mu opioid receptor agonist D-Ala2-N-methyl-Phe4-Gly5-ol-enkephalin (DAMGO, 0.7 microgram/5 microliters i.c.v.) was also attenuated in rats pretreated with U50,488H, U69,593 or PD117,302. These data indicate that activation of the kappa opioid receptor exerts an inhibitory effect on absence-like seizure activity in WAG/Rij rats.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗

PubMed1995

Seizure related changes in the regulation of opioid genes and transcription factors in the dentate gyrus of rat hippocampus.

An in situ hybridization study showed that limbic seizures induced by kainate strongly augmented the prodynorphin and proenkephalin messenger RNA levels in granular cells of the rat hippocampal dentate gyrus. Pentylenetetrazole increased the level of proenkephalin messenger RNA, but slightly decreased that of prodynorphin messenger RNA in the dentate gyrus. Administration of kainate to rats caused a profound increase in messenger RNAs of the transcription factor genes c-fos and c-jun in the dentate gyrus, followed by an increase in the level of the transcriptional complex activator protein-1 in hippocampal neurons. Pentylenetetrazole also elevated the formation of activator protein-1, but the effect appeared earlier than that induced by kainate. Thus, recurrent limbic seizures activate both prodynorphin and proenkephalin genes, whereas generalized clonic-tonic seizures seem to decrease the prodynorphin and increase the proenkephalin gene expression in the dentate gyrus. Furthermore, our present results suggest that the transcription factors, c-fos, c-jun and activator protein-1 complex may be involved in the process of inducing the hippocampal proenkephalin gene, while these factors might be differently involved in regulation of prodynorphin gene expression.

Animals↗

PubMed1995

Enhancing effect of staurosporine on NO production in rat peritoneal macrophages via a protein kinase C-independent mechanism.

Staurosporine (3-100 nM), frequently used as a protein kinase C (PKC) inhibitor, increased accumulation of nitrite in the culture medium of rat peritoneal macrophages up to 6 times above the control level. Moreover, when used in combination with the stable analogue of cyclic AMP, dibutyrylcyclic AMP (db cyclic AMP; 0.1 mM), and/or a cytokine, tumour necrosis factor-alpha (TNF alpha; 100 u ml-1), staurosporine synergistically potentiated, up to 30 times, nitrite accumulation. On the other hand, the other PKC inhibitors, calphostin C and H-7 (10 nM-10 microM) were not effective under the same conditions. The staurosporine-induced nitrite accumulation, in both the presence and the absence of TNF alpha and/or db cyclic AMP was effectively inhibited by the protein synthesis inhibitor, cycloheximide, or by the nitric oxide (NO) synthesis inhibitor, NG-monomethyl-L-arginine (L-NMMA). Thus our data suggest that staurosporine may enhance NO production in macrophages via intracellular mechanisms unrelated to the PKC inhibition.

Alkaloids↗

PubMed1994

The effects of cocaine-induced seizures on the proenkephalin mRNA level in the mouse hippocampus: a possible involvement of the nitric oxide pathway.

The effect of the nitric oxide synthase inhibitor L-NG-nitroarginine methyl ester (L-NAME, 2 and 10 mg/kg i.p.) on behavioral and biochemical changes induced by single and repeated administration of cocaine (45 mg/kg/day, i.p., for 5 days) was investigated in mice. Repeated cocaine produced a progressive increase in the intensity of seizures (a ca. 300% increase in the seizure score on day 5, as compared to day 1), this effect being associated with the enhancement of the proenkephalin mRNA level (ca. 240%) in the hippocampal dentate gyrus. L-NAME had no influence, but when used jointly with cocaine, it markedly attenuated both behavioral and biochemical effects of repeated cocaine. These data suggest that the nitric oxide pathway is involved in the progressive increase in excitability of the central nervous system after repeated cocaine administration.

Animals↗

PubMed1994

cAMP analogues and cholera toxin stimulate the accumulation of nitrite in rat peritoneal macrophage cultures.

Rat peritoneal macrophages incubated with the two stable analogues of cAMP, dibutyryl-cAMP and 8-bromo-cAMP, as well as with cholera toxin, released nitrite in a dose-dependent manner. Cholera toxin and dibutyryl-cAMP enhanced nitrite release induced by bacterial lipopolysaccharide. The stimulatory effects of all these substances were inhibited by the nitric oxide synthase inhibitor NG-monomethyl-L-arginine, dexamethasone and the protein synthesis inhibitor, cycloheximide. Our data indicate that the activation of cAMP-dependent pathway(s) can induce nitric oxide synthesis in rat peritoneal macrophages even in the absence of immunological stimuli, such as exogenous cytokines or lipopolysaccharide, although the exact mechanism of this phenomenon remains to be established.

8-Bromo Cyclic Adenosine Monophosphate↗

PubMed1994

Effects of mu and delta opioid receptor agonists and antagonists on absence epilepsy in WAG/Rij rats.

The effects of various types of opioid receptor agonists and antagonists were determined in a genetic rat model for generalized absence epilepsy. Rats of the WAG/Rij strain spontaneously showed several hundred spike-wave discharges per day. Intracerebroventricular (i.c.v.) injections of the selective mu agonist DAMGO (0.2, 0.7 microgram) resulted in a dose-related increase in the number of spike-wave discharges, while the selective delta agonist DPDPE (20, 60 micrograms) was without effect. DAMGO reduced the duration of automatic behavior, enhanced the immobile behavior (after the low dose) and had no effect on exploratory behavior. On the other hand, DPDPE significantly enhanced the total time spent on exploration, but did not influence other behavioral parameters. There was no correlation between the ability to the drug to modulate the epileptic activity and behavioral changes. The nonselective antagonist naloxone, administered either i.p. (0.4, 2.0, 10 mg/kg) or i.c.v. (10, 50 micrograms), increased the number of spike-wave discharges in a dose-dependent way. The specific delta receptor antagonist naltrindole (0.3, 1 mg/kg) was without effect, as was the irreversible mu receptor antagonist beta-funaltrexamine (beta-FNA). Pretreatment with beta-FNA diminished the action of DAMGO. These results clearly indicate that activation of the mu opioid receptor increases the number of spike-wave discharges, and that modulation of delta receptors is not effective. On the other hand, the naloxone-induced enhancement of spike-wave discharges, suggests a tonic control of the epileptic activity by another opioid system. These results point to an important role of the mu-, but not delta-, receptor in facilitation of the epileptic activity in WAG/Rij rats.

Analysis of Variance↗

PubMed1994

Inhibition of nitric oxide synthase attenuates the development of morphine tolerance and dependence in mice.

The effect of the nitric oxide (NO) synthase inhibitor L-NG-nitroarginine methyl ester (L-NAME, 5-20 mg/kg i.p.) and NG-nitro-L-arginine (NO2Arg, 5-20 mg/kg i.p.) on morphine-induced analgesia, as well as on morphine induced tolerance and dependence was examined in male albino Swiss mice. Neither acute nor repeated (for 5 days) administration of the nitric oxide synthase inhibitor, L-NAME affected the morphine induced analgesia, as measured by hot plate and tail-flick tests. On the other hand, administration of L-NAME or NO2Arg along with morphine prevented the development of tolerance to the analgesic effect of morphine for at least 7 days, whereas the analgesic effect of morphine alone disappeared after 5 days. L-NAME and NO2Arg also attenuated some signs of morphine dependence, as assessed by naloxone (2 mg/kg)-precipitated withdrawal. These results indicate that NO may play a role in the development of morphine tolerance and dependence.

Amino Acid Oxidoreductases↗

PubMed1994

Seizure-induced expression of G proteins in the rat hippocampus.

The effects of kainic acid (15 mg/kg i.p.) on alpha subunits of the Gs and Go protein mRNA levels in the rat hippocampal formation were investigated. An in situ hybridization study showed an increase in the Gs alpha mRNA level in the dentate gyrus at 3 h (by ca. 17%), 24 h (by ca. 75%), 72 h (by ca. 89%) and 30 days (by ca. 59%) after kainic acid administration. An emulsion autoradiography revealed enhancement in the Gs alpha mRNA signal intensity over granular cells of the dentate gyrus and over some hilar cells adjacent to the granule cell layer, most likely in GABA interneurons. The Gs alpha mRNA showed a slight tendency to increase in the CA1 and CA3 pyramidal cell layers at 3 h after kainic acid administration, but it decreased after 24 h, 72 h and 30 days. The latter decrease correlated well with the pyramidal cells loss in those areas. Kainic acid differently influenced the Go alpha mRNA level in the dentate gyrus: it had no effect after 3 h, while after 24 h the mRNA level tended to decrease (by ca. 16%); then it increased after 72 h (by ca. 20%) and, to a lesser extent, after 30 days (by ca. 12%). The Go alpha mRNA level in CA1 and CA3 tended to decrease at 3 h after kainic acid administration; the signal completely disappeared after 24, 72 h as well as after 30 days.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1994

The effect of chronic morphine and cocaine administration on the Gs and Go protein messenger RNA levels in the rat hippocampus.

The effects of single (20 or 50 mg/kg) and repeated morphine (14 days, rising doses: four days, 2 x 10 mg/kg; two days, 2 x 20 mg/kg; two days, 2 x 30 mg/kg; two days, 2 x 40 mg/kg and four days, 2 x 50 mg/kg i.p.) or cocaine (20 mg/kg i.p., once every hour for 3 h for one or five days) on the Gs alpha and Go alpha protein messenger RNA levels were investigated in the rat hippocampus. As shown by in situ hybridization immunocytochemistry and optical densitometry of autoradiograms, acute morphine (20 mg/kg) decreased the Gs alpha messenger RNA level and increased the Go alpha messenger RNA level in the dentate gyrus and CA1 hippocampal region. Repeated morphine administration significantly decreased the hippocampal Gs alpha messenger RNA level at 12, 24 and 48 h, and enhanced the Go alpha messenger RNA level in those areas at 12 and 24 h after the last injection. Repeated and acute cocaine administration decreased the Go alpha messenger RNA level in the CA3 region and increased it in the CA1 at various time points. Furthermore, in all hippocampal regions, acute, but not chronic, cocaine administration increased the Gs alpha mRNA level at 24 h, whereas chronic treatment significantly decreased that level at 48 h after the last dose. The data indicate that chronic morphine elevates the Go and attenuates the Gs protein biosynthesis at later time points in all regions of the hippocampal formation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1994

Repeated ethanol differently affects opioid peptide biosynthesis in the rat pituitary.

In situ hybridization and specific radioimmunoassays were used to study the influence of ethanol on proopiomelanocortin (POMC) and prodynorphin (PDYN) biosynthesis in the rat pituitary. Repeated intragastric ethanol administration (starting with a total daily dose of 5 g/kg every 2nd day, until a dose of 10 g/kg was attained on the 10th day and that dose was maintained by the 19th day) resulted in a reduction in the POMC mRNA level (about 20%) in the intermediate lobe of the pituitary (3 h after the last dose), whereas the level of beta-endorphin in the neurointermediate lobe was attenuated (by about 32%) only during the withdrawal (48 h after the last dose). Additionally, the plasma level of beta-endorphin after repeated ethanol and during withdrawal was significantly reduced (by about 44 and 66%, respectively). No changes in the POMC mRNA or the beta-endorphin levels were detected in the anterior lobe. In contrast, the PDYN mRNA level was found to be decreased in the anterior lobe during the withdrawal (by about 43%). This decrease was in conjunction with an increase in the alpha-neoendorphin level (by about 57%) in that lobe. The PDYN mRNA level in the intermediate lobe and the alpha-neoendorphin level in the neurointermediate lobe were unchanged after ethanol, as well as during the withdrawal period. Acute ethanol (5 g/kg) decreased the level of beta-endorphin in the anterior lobe; this effect being associated with an elevation in the peptide level in plasma. On the other hand, acute ethanol had no effect on the POMC and PDYN mRNA levels, nor did it affect the alpha-neoendorphin concentration in the pituitary.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1994

Influence of SIN-1 and sodium nitroprusside (NANP) on ox-LDL metabolism in macrophages.

Effects of NO-donors (3-morpholinosydnonimine-SIN-1 and sodium nitroprusside NaNP) on the accumulation and degradation of oxidized LDL (ox-LDL) by macrophages were studied. Ox-LDL, but not native-LDL (n-LDL) suppressed the LPS-stimulated biosynthesis of NO by macrophages. SIN-1 at low concentrations < 100 microM was without any effect while SIN-1 at high concentration (300 microM) and NaNP (30-300 microM) stimulated the accumulation and degradation of ox-LDL by macrophages. The pretreatment of macrophages with NG-monomethyl-L-arginine (L-NMMA, 3 microM) for 24 hours had the same stimulatory effect. The inhibition of endogenous formation of NO, by L-NMMA profoundly changed the pattern of action of NO-donors on ox-LDL catabolism by macrophages; the stimulatory action of SIN-1 was transformed to the inhibitory action on the accumulation and degradation of ox-LDL whereas NaNP lost its stimulatory action entirely. Our interpretation of this unexpected interactions between SIN-1, NaNP and L-NMMA is as follows. Endogenous NO in macrophages inhibits the accumulation of ox-LDL and therefore, the stimulatory effect of L-NMMA has been overcome by exogenous NO from SIN-1. However, NO at high concentrations promotes lipid accumulation in macrophages and thereby, in the absence of L-NMMA, SIN-1 at high concentrations and NaNP produced a paradoxical stimulatory effect in macrophages. NaNP is not a proper NO-donor and its mode of action differed from that of SIN-1. In conclusion, NO at low physiological concentrations keeps scavenger receptors of macrophages downregulated and hence endogenous NO may show anti-atherogenic properties.

Animals↗

PubMed1993

Inhibition of nitric oxide synthase enhances morphine antinociception in the rat spinal cord.

NG-nitro-L-arginine methyl ester (L-NAME, 400-1500 micrograms), administered intrathecally (ith.), elicits a slight but dose-related antinociception in rats, assessed by tail-flick and paw pressure tests. L-NAME (400 micrograms) and morphine (0.5 microgram) coadministered ith. elicit a profound and long-lasting antinociception, which is abolished by ith. administration of 3-morpholino-sydnonimine (SIN-1, 100 micrograms). Hemoglobin (266 micrograms) administered ith. also slightly potentiates morphine antinociception. These results suggest that nitric oxide (NO) is involved in spinal nociceptive events, and that the increased production of NO following the nociceptive input may diminish the efficiency of opioid antinociception in the spinal cord.

Amino Acid Oxidoreductases↗

PubMed1992

Repeated ethanol administration decreases prodynorphin biosynthesis in the rat hippocampus.

The effect of single and repeated (19 days) intragastric ethanol administration on prodynorphin mRNA and alpha-neoendorphin levels in the hippocampus of the rat was studied using in situ hybridization and RIA methods, respectively. Single ethanol administration had no effect on either of these two parameters. Repeated ethanol administration decreased both immunoreactive (ir) alpha-neoendorphin (by ca. 37%) and prodynorphin mRNA (by ca. 57%) levels. Two days after the last dose of ethanol no changes in the ir-alpha-neoendorphin tissue content were found, whereas the level of prodynorphin mRNA remained decreased (by ca. 44%). These results suggest that repeated ethanol evokes a long-lasting decrease in the biosynthesis of hippocampal prodynorphin, this effect that may play some role in the ethanol-induced deficiency of neuronal functions.

Alcoholism↗

PubMed1992

Differential effects of opioid receptor agonists on nociception and cAMP level in the spinal cord of monoarthritic rats.

Changes in functional responsiveness of spinal opioid receptors in monoarthritic rats were investigated at the behavioral and the molecular level. After intrathecal administration of morphine, D-Ala2-D-Leu5-enkephalin (DADLE), D-Pen2-D-Pen5-enkephalin (DPDPE) and dynorphin monoarthritic rats showed an enhanced antinociceptive response as measured by a tail-flick latency. No such changes were observed following administration of the selective kappa agonists U50,488H and U69,593. The opioid mu and delta receptor agonists (0.1-1.0 microM) inhibited the basal, as well as the forskolin-stimulated cAMP formation in spinal cord slices obtained from monoarthritic rats, whereas no significant changes were found in control animals. Higher concentrations of the mu and delta opioid receptor agonists were required to attenuate the cAMP level in spinal cord of control animals. The selective kappa agonists U50,488H and U69,593 did not influence the cAMP formation in monoarthritic or control animals. Additionally, we found that the GppNHp-stimulated level of cAMP was higher in the spinal cord slices of monoarthritic rats, which points to an enhanced responsiveness of the adenylate cyclase effector system to the action of this GTP analog. Our data suggest that the enhanced antinociceptive response to intrathecally administered opioids in monoarthritic rats may be connected with the increased sensitivity of adenylate cyclase to the inhibitory effects of mu and delta agonists.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗

PubMed1992

Endogenous opioid peptides in brain and pituitary of rats with absence epilepsy.

The level of opioid peptides in several brain areas and in the pituitary was estimated in WAG/Rij rats, which are considered to be a genetic animal model for human absence epilepsy. In comparison with three groups of non-epileptic controls, these epileptic rats had an elevated level of the proenkephalin-derived peptide Met-enkephalin-Arg6-Gly7-Leu8 in the mesencephalon and striatum, while the level of the prodynorphin-derived peptide alpha-neoendorphin was increased in the striatum and hippocampus. In addition various age- and/or strain-related changes in these peptide levels were found in the hippocampus, thalamus, striatum, frontal cortex and neurointermediate lobe of the pituitary. No difference in the hypothalamic beta-endorphin level were found between epileptic and non-epileptic rats, though strain- and/or age-related changes in the peptide content were detected in both lobes of the pituitary. The increased level of proenkephalin and prodynorphin opioid peptides in brain structures, essential for the appearance of spike-wave discharges, suggests that these opioid systems, but not proopiomelanocortin one, may play a role in absence epilepsy.

Animals↗

PubMed1992

The effects of excitatory amino acids on proenkephalin and prodynorphin mRNA levels in the hippocampal dentate gyrus of the rat; an in situ hybridization study.

Injection of N-methyl-D-aspartate (NMDA, 7.5 micrograms) kainate (1 microgram) or quisqualate (2 micrograms) into the rat dorsal hippocampus induced wet-dog shakes and convulsions. As shown by an in situ immunohistochemical analysis, 3 h after the excitatory amino acids injections the rats displayed a bilateral profound elevation of the proenkephalin and prodynorphin mRNA levels in dentate gyrus granule cells (2-3 or 1.5-2 fold higher than control levels, respectively). Pretreatment of rats with D-amino-phosphonovalerate (D-APV, 10 micrograms), a selective antagonist of NMDA receptor, prevented the behavioral and biochemical changes evoked by NMDA. The changes in the behavior and gene expression evoked by kainate or quisqualate were diminished in rats which received 6-cyano-7-nitroquinoxaline-2,3-dion (CNQX, 2 micrograms), a putative antagonist of quisqualate and kainate receptors. The study demonstrated that activation of NMDA, quisqualate or kainate receptors in the hippocampus induced seizures associated with a marked increase in the proenkephalin (PENK) and the prodynorphin (PDYN) gene expression in the rat dentate gyrus.

2-Amino-5-phosphonovalerate↗

PubMed1992

Time-dependent changes in the activity of opioid systems in the spinal cord of monoarthritic rats--a release and in situ hybridization study.

The activity of the endogenous opioid systems was analysed in the spinal cord of rats 12 h, one, five or 14 days after injection of the Freund Adjuvant into the hind limb sole. The tissue level of immunoreactive Met-enkephalin-Arg6-Gly7-Leu8, a peptide derived from proenkephalin, started to rise 12 h after Freund Adjuvant inoculation and remained enhanced until day 14. The spontaneous release of the immunoreactive Met-enkephalin-Arg6-Gly7-Leu8 was also elevated at all the examined time points, the peak occurring on day 1. No changes were observed in the stimulated release except on day 14, when the peptide release was decreased. The proenkephalin messenger RNA level was enhanced at all the time points on the ipsilateral side of the spinal cord in laminae I-II, whereas in lamina V an increase was observed only on days 1 and 5. An increase in the proenkephalin messenger RNA level on the contralateral side was observed only in laminae I-II and only on days 1 and 5. The tissue level of immunoreactive alpha-neoendorphin, a peptide derived from prodynorphin, was significantly increased on days 5 and 14. The spontaneous immunoreactive alpha-neoendorphin release from spinal cord slices was elevated at all the time points studied, whereas the stimulated release of the peptide was strongly increased 12 h after Freund Adjuvant inoculation but gradually declined on the following days. An in situ hybridization study showed that the prodynorphin messenger RNA level in laminae I-II was increased at all the examined time points.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗