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Limitations on the use of animals for biological experiments has forced a search for alternative simpler organisms for testing the biological activity of organic com­pounds. The aim of this research was to use Tetrahymena termophila for testing the opioid activity of pep­tide, -casomorphin-7, isolated from cheese. Three kinds of test were used: 1) test for in­hi­bition of protozoa mobility, 2) trap test, and 3) phagocytose test. The results for the trap test suggests that it can be used to determine the presence of harmful substances occurring in food. Determination of the opioid properties using the phagocytose test showed that morphine inhibits the phagocytose process of Tetrahymena termophila. -casomorphin-7 had smaller inhibitory effect on ph­ago­cy­to­sis than mor­phine. it can be said that our results and literature prove the possibility of using protozoa Tet­rahy­mena termophila for studies of antinutritional sub­stances and opioids in raw and food products.
Respiratory effects of morphine injection to the femoral vein were investigated in urethane and chloralose anaesthetized and spontaneously breathing rats, prior to and after midcervical vagotomy. Bolus injection of morphine HCl at a dose of 2 mg/kg of body weight induced depression of ventilation in all rats, due to the significant decrease in tidal volume and to the decline in respiratory rate both pre- and post-vagotomy. Expiratory apnoea of mean duration of 10.0±3.4 s was present in the vagally intact rats only. Bilateral midcervical section of the vagus nerve precluded the occurrence of apnoea. Prolongation of the expiratory time (TE morphine / TE control), which amounted to10.7±2.2-fold in the intact state, was apparently reduced to 1.5±0.3-fold after division of the vagi. Morphine significantly decreased mean arterial pressure (MAP) at 30 s after the challenge, the effect persisted for not less than 1 minute and was absent in vagotomized rats. The respiratory changes evoked by morphine reverted to the control level after intravenous injection of naloxone at a dose of 1 mg/kg. Results of this study indicate that opioid receptors on vagal afferents are responsible for the occurrence of apnoea and hypotension evoked by morphine.
In previous studies the effect of endogenous opioid peptides (EOP) on LH secretion was mainly considered at the hypothalamic level, while opioid involvement in the modulation of LH secretion at the pituitary level remains insufficiently elucidated. Therefore, the present study was undertaken to determine the expression of genes encoding opioid precursors – proopiomelanocortin (POMC), proenkephalin (PENK), prodynorphin (PDYN) and opioid receptors – mu, delta, kappa in the porcine anterior pituitary throughout the estrous cycle. Additionally, the mRNA content of ß-LH subunit and GnRH receptor (GnRH-R) was estimated. Pituitaries (5×N = 7) were collected from sows on days 3-5, 8-10, 13-15, 16-17 and 19-20 of the cycle and gene expression was determined using a semi-quantitative RT-PCR assay. The expression of POMC, PDYN, delta and kappa receptor genes was variable across the cycle, whereas the expression of PENK and mu receptor genes remained relatively stable. The POMC mRNA content was the lowest on days 19-20 of the cycle and the PDYN content was reduced on days 8-10. The delta receptor mRNA content was elevated on days 3-5, while the kappa receptor mRNA content was decreasing over the luteal phase. Changes in the expression of genes encoding ß-LH and GnRH-R were also demonstrated. These results indicate variable activity of pituitary opioid systems in cyclic pigs and suggest implication of EOP in the modulation of LH secretion at the pituitary level.
Lifetime distribution analysis were performed to study the influence of Leu configura­tion in position 5 on changes of the peptide chain of cyclic analogues of enkephalins con­taining a fluorescence donor and acceptor in different solvents. The configuration change of Leu5 in all the analogues of enkephalins studied which contain donor-acceptor pairs has no apparent influence on Trp lifetime distributions. In contrast, there is a significant solvent effect on the shape of lifetime distribution.
A series of new nociceptin analogues containing cysteine was tested for their noci­ceptive effects in tail-flick test on mice after icv injection. The cysteines were intro­duced in order to get irreversibly binding analogues based on the assumption that the cysteines in the ligand can interact with the cysteines from the receptor to form an S-S bridge. In vivo tests revealed that CysX-nociceptin (1-13)-NH2 (Cys1-NC) is an antagonist, whereas Cys7-NC is an agonist. Gly1 ,[Phe(p-NO2)]4 -NC was less active indi­cating that the antagonist properties of Cys1-NC are associated with the presence of the sulfhydryl group of cysteine. The analogues D-Cys2 and Cys3 were also almost in­active.
Opioids administered by intracerebroventricular injections produce analgesic responses in rats. The present study was undertaken to investigate the effects of a highly selective m-opioid receptor ligand morphiceptin on trigemino-hypoglossal reflex in rats. The analgesic effect of morphiceptin was compared with another opioid peptide, Met-enkephalin. With the experimental settings used in this study, we have demonstrated that both morphiceptin and Met-enkephalin show significant dose-dependent analgesic effects after i.c.v. administration in rats as assayed by trigemino-hypoglossal reflex test. The antinociceptive response to Met-enkephalin was short lasting and was observed 10 to 15 min after i.c.v. perfusion. Morphiceptin had a relatively longer duration of antinociceptive action, the effect was observed 20- 50 min after i.c.v. perfusion. Neither morphiceptin nor Met-enkephalin produced antinociception after peripheral injections. The results of the present study indicate that both tested peptides act at µ-opioid receptors situated in the central nervous system. They also suggest that µ-opioid receptors present in the central nervous system are an important element of the trigemino-hypoglossal reflex arc. For that reason selective m-opioid receptor ligands, like morphiceptin, inhibit the reflex more significantly.
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