Protracted opioid withdrawal is considered to be a traumatic event with many adverse effects. However, little attention is paid to its consequences on the protein expression in the rat brain. A better understanding of the changes at the molecular level is essential for designing future innovative drug therapies. Our previous proteomic data indicated that long-term morphine withdrawal is associated with altered proteins functionally involved in energy metabolism, cytoskeletal changes, oxidative stress, apoptosis, or signal transduction. In this study, we selected peroxiredoxin II (PRX II) as a marker of oxidative stress, 14-3-3 proteins as adaptors, and creatine kinase-B (CK-B) as a marker of energy metabolism to detect their amounts in the brain cortex and hippocampus isolated from rats after 3-month (3 MW) and 6-month morphine withdrawal (6 MW). Methodically, our work was based on immunoblotting accompanied by 2D resolution of PRX II and 14-3-3 proteins. Our results demonstrate significant upregulation of PRX II in the rat brain cortex (3-fold) and hippocampus (1.3-fold) after 3-month morphine abstinence, which returned to the baseline six months since the drug was withdrawn. Interestingly, the level of 14-3-3 proteins was downregulated in both brain areas in 3 MW samples and remained decreased only in the brain cortex of 6 MW. Our findings suggest that the rat brain cortex and hippocampus exhibit the oxidative stress-induced vulnerability represented by compensatory upregulation of PRX II after three months of morphine withdrawal.

• MeSH
• abstinenční syndrom * metabolismus   MeSH
• hipokampus metabolismus   MeSH
• krysa rodu rattus MeSH
• morfin metabolismus   MeSH
• mozek metabolismus   MeSH
• peroxiredoxiny metabolismus   farmakologie   MeSH
• proteiny 14-3-3 metabolismus   MeSH
• proteomika MeSH
• upregulace MeSH
• závislost na morfiu * MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Early-life morphine exposure causes a variety of behavioural and physiological alterations observed later in life. In the present study, we investigated the effects of prenatal and early postnatal morphine on the maturation of the circadian clockwork in the suprachiasmatic nucleus and the liver, and the rhythm in aralkylamine N-acetyltransferase activity in the pineal gland. Our data suggest that the most affected animals were those born to control, untreated mothers and cross-fostered by morphine-exposed dams. These animals showed the highest mesor and amplitude in the rhythm of Per2, Nr1d1 but not Per1 gene expression in the suprachiasmatic nuclei (SCN) and arrhythmicity in AA-NAT activity in the pineal gland. In a similar pattern to the rhythm of Per2 expression in the SCN, they also expressed Per2 in a higher amplitude rhythm in the liver. Five of seven specific genes in the liver showed significant differences between groups in their expression. A comparison of mean relative mRNA levels suggests that this variability was caused mostly by cross-fostering, animals born to morphine-exposed dams that were cross-fostered by control mothers and vice versa differed from both groups of natural mothers raising offspring. Our data reveal that the circadian system responds to early-life morphine administration with significant changes in clock gene expression profiles both in the SCN and in the liver. The observed differences between the groups suggest that the dose, timing and accompanying stress events such as cross-fostering may play a role in the final magnitude of the physiological challenge that opioids bring to the developing circadian clock.

• MeSH
• cirkadiánní hodiny * MeSH
• cirkadiánní rytmus MeSH
• krysa rodu rattus MeSH
• laktace MeSH
• morfin metabolismus   farmakologie   MeSH
• nucleus suprachiasmaticus metabolismus   MeSH
• těhotenství MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Integrated behavioral paradigms such as nociceptive processing coupled to anti-nociceptive responsiveness include systemically-mediated states of alertness, vigilance, motivation, and avoidance. Within a historical and cultural context, opium and its biologically active compounds, codeine and morphine, have been widely used as frontline anti-nociceptive agents. In eukaryotic cells, opiate alkaloids and opioid peptides were evolutionarily fashioned as regulatory factors in neuroimmune, vascular immune, and systemic immune communication and auto-immunoregulation. The significance of opioidergic regulation of immune function was validated by the identification of novel μ and δ opioid receptors on circulating leukocytes. The novel μ3 opioid receptor subtype has been characterized as an opioid peptide-insensitive and opiate alkaloid-selective G protein-coupled receptor (GPCR) that is functionally linked to the activation of constitutive nitric oxide synthase (cNOS). Opioid peptides stimulate granulocyte and immunocyte activation and chemotaxis via activation of a novel leukocyte δ2 receptor subtype. However, opiate alkaloid μ3 receptor agonists inhibit these same cellular activities. Opiate coupling to cNOS and subsequent production and release of mitochondrial nitric oxide (NO) suggests an evolutionary linkage to similar physiological events in prokaryotic cells. A subpopulation of immunocytes from Mytilus edulis and Leucophaea maderae and human granulocytes respond to low opioid concentrations, mediated by the adherence-promoting role of (D-Ala2-D-Met5)-enkephalinamide (DAMA), which is blocked by naloxone in a dose-dependent manner. Neutral endopeptidase 24.11 (NEP), or enkephalinase (CD10), is present on both human and invertebrate immunocytes. Alkaloids, including morphine, are found in both prokaryotic and eukaryotic cells and may have evolved much later in evolution through horizontal gene transfer. It is possible that opioid-mediated regulatory activities were conserved and elaborated during evolution as the central nervous system (CNS) became immunologically isolated by the blood-brain barrier. Thus, opioid receptor coupling became significant for cognitive and behavioural processes. Although opioid peptides and alkaloids work synergistically to suppress nociception, they mediate different actions in immune surveillance. Increased understanding of the evolutionary development of opioid receptors, nociceptive and anti-nociceptive pathways, and immunomodulation may help in the understanding of the development of tolerance to the clinical use of opiates for pain management. The significance of endogenous morphine's importance to evolution can be ascertained by the number of physiological tissues and systems that can be affected by this chemical messenger mechanism, which transcends pain. An integrated review is presented of opioid and opiate receptors, immunomodulation, and pain associated with inflammation, from an evolutionary perspective.

• MeSH
• biologická evoluce MeSH
• bolest metabolismus   MeSH
• chování MeSH
• imunita MeSH
• imunomodulace MeSH
• lidé MeSH
• morfin metabolismus   MeSH
• opioidní peptidy metabolismus   MeSH
• oxid dusnatý metabolismus   MeSH
• receptory opiátové genetika   metabolismus   MeSH
• zánět metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

The interactions between TRPV1 and µ-opioid receptors (MOR) have recently attracted much attention because these two receptors play important roles in pain pathways and can apparently modulate each other's functioning. However, the knowledge about signaling interactions and crosstalk between these two receptors is still limited. In this study, we investigated the mutual interactions between MOR and TRPV1 shortly after their activation in HEK293 cells expressing these two receptors. After activation of one receptor we observed significant changes in the other receptor's lateral mobility and vice versa. However, the changes in receptor movement within the plasma membrane were not connected with activation of the other receptor. We also observed that plasma membrane β-arrestin 2 levels were altered after treatment with agonists of both these receptors. Knockdown of β-arrestin 2 blocked all changes in the lateral mobility of both receptors. Furthermore, we found that β-arrestin 2 can play an important role in modulating the effectiveness of ERK1/2 phosphorylation after activation of MOR in the presence of TRPV1. These data suggest that β-arrestin 2 and ERK1/2 are important mediators between these two receptors and their signaling pathways. Collectively, MOR and TRPV1 can mutually affect each other's behavior and β-arrestin 2 apparently plays a key role in the bidirectional crosstalk between these two receptors in the plasma membrane.

• MeSH
• arrestiny metabolismus   MeSH
• beta arrestin 2 metabolismus   fyziologie   MeSH
• beta arrestiny metabolismus   MeSH
• buněčná membrána metabolismus   fyziologie   MeSH
• fosforylace MeSH
• HEK293 buňky MeSH
• kationtové kanály TRPV metabolismus   fyziologie   MeSH
• lidé MeSH
• MAP kinasový signální systém fyziologie   MeSH
• morfin metabolismus   MeSH
• opioidní analgetika metabolismus   MeSH
• receptory opiátové mu metabolismus   fyziologie   MeSH
• receptory opiátové metabolismus   MeSH
• signální transdukce MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

• MeSH
• asfyxie diagnóza   MeSH
• biometrie metody   MeSH
• dopravní nehody MeSH
• ethanol analýza   otrava   MeSH
• inzulin analýza   otrava   MeSH
• iris anatomie a histologie   MeSH
• lidé MeSH
• morfin analýza   metabolismus   MeSH
• mydriáza chemicky indukované   MeSH
• patologický nystagmus chemicky indukované   MeSH
• pití alkoholu patofyziologie   škodlivé účinky   MeSH
• sklivec chemie   metabolismus   účinky léků   MeSH
• soudní toxikologie * MeSH
• Check Tag
• lidé MeSH

There are some indications that biased μ-opioid ligands may diversely affect μ-opioid receptor (MOR) properties. Here, we used confocal fluorescence recovery after photobleaching (FRAP) to study the regulation by different MOR agonists of receptor movement within the plasma membrane of HEK293 cells stably expressing a functional yellow fluorescent protein (YFP)-tagged μ-opioid receptor (MOR-YFP). We found that the lateral mobility of MOR-YFP was increased by (D-Ala(2),N-MePhe(4),Gly(5)-ol)-enkephalin (DAMGO) and to a lesser extent also by morphine but decreased by endomorphin-2. Interestingly, cholesterol depletion strongly enhanced the ability of morphine to elevate receptor mobility but significantly reduced or even eliminated the effect of DAMGO and endomorphin-2, respectively. Moreover, the ability of DAMGO and endomorphin-2 to influence MOR-YFP movement was diminished by pertussis toxin treatment. The results obtained by agonist-stimulated [(35)S]GTPγS binding assays indicated that DAMGO exhibited higher efficacy than morphine and endomorphin-2 did and that the efficacy of DAMGO, contrary to the latter agonists, was enhanced by cholesterol depletion. Overall, our study provides clear evidence that biased MOR agonists diversely affect receptor mobility in plasma membranes as well as MOR/G protein coupling and that the regulatory effect of different ligands depends on the membrane cholesterol content. These findings help to delineate the fundamental properties of MOR regarding their interaction with biased MOR ligands and cognate G proteins.

• MeSH
• bakteriální proteiny genetika   metabolismus   MeSH
• buněčná membrána účinky léků   metabolismus   MeSH
• cholesterol nedostatek   MeSH
• enkefalin, Ala(2)-MePhe(4)-Gly(5)- metabolismus   farmakologie   MeSH
• FRAP MeSH
• guanosin 5'-O-(3-thiotrifosfát) metabolismus   MeSH
• HEK293 buňky MeSH
• konfokální mikroskopie MeSH
• lidé MeSH
• ligandy MeSH
• luminescentní proteiny genetika   metabolismus   MeSH
• morfin metabolismus   farmakologie   MeSH
• narkotika - antagonisté farmakologie   MeSH
• oligopeptidy metabolismus   farmakologie   MeSH
• pertusový toxin farmakologie   MeSH
• proteiny vázající GTP - alfa-podjednotky Gi-Go metabolismus   MeSH
• receptory opiátové mu agonisté   genetika   metabolismus   MeSH
• rekombinantní fúzní proteiny metabolismus   MeSH
• transfekce MeSH
• transport proteinů MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

• Klíčová slova
• genetický polymorfizmus, aktivní/toxické metabolity, opioidy,
• MeSH
• cytochrom P-450 CYP2D6 farmakokinetika   metabolismus   MeSH
• játra enzymologie   metabolismus   MeSH
• kodein metabolismus   MeSH
• lékové interakce MeSH
• lidé MeSH
• morfin metabolismus   MeSH
• opioidní analgetika aplikace a dávkování   farmakokinetika   metabolismus   MeSH
• oxykodon analogy a deriváty   metabolismus   MeSH
• polymorfismus genetický genetika   účinky léků   MeSH
• tramadol metabolismus   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• Check Tag
• lidé MeSH

Práce uvádí toxikologické charakteristiky 198 případů akutní parenterální intoxikace heroinem, analyzuje klinicky získaný rozsah hladin jeho metabolitů v krvi a v moči. Jsou objasněny hlavní příčiny smrti u hospitalizovaných obětí otravy heroinem. Je určen vztah pravděpodobnosti úmrtí při zjištění morfinu v tělních tekutinách obětí, byly stanoveny jeho koncentrace v krvi a v moči, které nepochybně ukazují na to, že nastane smrt způsobená otravou. Bylo stanoveno, že smrt způsobená otravou heroinem může nastat v celém rozmezí jeho detekovatelných hladin. Není pochyb o tom, že koncentrace morfinu v krvi vyšší než 2,0 µg/ml by měla být považována za fatální.

The paper presents toxicological characteristics of 198 cases of acute parenteral heroin intoxication, analyzes the clinically encountered range of blood and urinary concentrations of its metabolites. The principal causes of death are elucidated in victims of heroin poisoning at the hospital stage. Where there is a relationship of death probability to the detection of morphine in the victims' biological fluids is considered; its blood and urinary concentrations are determined, which undoubtedly suggests the occurrence of poisoning-related death. It has been established that death from poisoning by heroin may occur in the whole range of its detectable concentrations. There is no doubt that the blood morphine concentrations of at least 2.0 µg/ml should be considered to be fatal.

• MeSH
• heroin farmakologie   toxicita   MeSH
• lidé MeSH
• morfin aplikace a dávkování   kontraindikace   metabolismus   MeSH
• mortalita MeSH
• toxikologie metody   MeSH
• Check Tag
• lidé MeSH

V práci jsou uvedeny toxikologické charakteristiky 198 případů akutní parenterální intoxikace heroinem a úplný seznam koncentrací jejich metabolitů, které se používají v praxi. Byl učiněn pokus stanovit riziko úmrtí při určitých koncentracích morfinu v krvi u akutní intoxikace heroinem v závislosti na věku a pohlaví. Bylo zjištěno, že střední smrtná koncentrace morfinu (LD50) se významně liší u mužů (0,78 µg/ml) a u žen (0,98 µg/ml) a u osob mladších (0,39 µg/ml) a starších (1,50 µg/ml) 25 let. Při věku nižším než 25 let, většina obětí vykazuje téměř totožné reakce v úzkém rozmezí koncentrací (LD50 = 0,39 µg/ml; LD100 = 1,57 µg/ml). U osob starších než 25 let jsou velké individuální rozdíly v jejich citlivosti na koncentraci toxické látky (LD50 = 1,50 µg/ml; LD100 = 3,90 µg/ml).

The paper presents the toxicological characteristics of 198 cases of acute parenteral heroin intoxication in the whole range of concentrations of their metabolites, which are encountered in practice. An attempt was made to estimate the risk of death within possible blood morphine concentrations in acute heroin intoxication, by taking into account gender- and age-related differences. The median lethal morphine concentration (LD50) has been found to greatly differ in males (0.78 µg/ml) and females (0.98 µg/ml) and in those aged under (0.39 µg/ml) and above (1.50 µg/ml) 25 years. At the age under 25 years, most victims show nearly the same response in the narrow range of concentrations (LD50 = 0.39 µg/ml; LD100 = 1.57 µg/ml); in persons above 25 years of age, there are great individual differences in their sensitivity to the concentration of a toxic substance (LD50 = 1.50 µg/ml; LD100 = 3.90 µg/ml).

• MeSH
• heroin krev   moč   otrava   MeSH
• lidé MeSH
• morfin kontraindikace   metabolismus   normy   MeSH
• rozložení podle pohlaví MeSH
• věkové faktory MeSH
• Check Tag
• lidé MeSH

• MeSH
• analýza moči MeSH
• dospělí MeSH
• lidé MeSH
• morfin metabolismus   MeSH
• Papaveraceae MeSH
• poruchy spojené s užíváním opiátů diagnóza   MeSH
• potraviny MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• přehledy MeSH
• srovnávací studie MeSH