RESUMO
Thalidomide is an antiinflammatory, antiangiogenic and immunomodulatory agent which has been used for the treatment of erythema nodosum leprosum and multiple myeloma. It has also been employed in treating complex regional pain syndromes. The current study aimed to reveal the molecular mechanisms underlying thalidomide-induced pain antihypersensitive effects in neuropathic pain. Thalidomide gavage, but not its more potent analogs lenalidomide and pomalidomide, inhibited mechanical allodynia and thermal hyperalgesia in neuropathic pain rats induced by tight ligation of spinal nerves, with ED50 values of 44.9 and 23.5 mg/kg, and Emax values of 74% and 84% MPE respectively. Intrathecal injection of thalidomide also inhibited mechanical allodynia and thermal hyperalgesia in neuropathic pain. Treatment with thalidomide, lenalidomide and pomalidomide reduced peripheral nerve injury-induced proinflammatory cytokines (TNFα, IL-1ß and IL-6) in the ipsilateral spinal cords of neuropathic rats and LPS-treated primary microglial cells. In contrast, treatment with thalidomide, but not lenalidomide or pomalidomide, stimulated spinal expressions of IL-10 and ß-endorphin in neuropathic rats. Particularly, thalidomide specifically stimulated IL-10 and ß-endorphin expressions in microglia but not astrocytes or neurons. Furthermore, pretreatment with the IL-10 antibody blocked upregulation of ß-endorphin in neuropathic rats and cultured microglial cells, whereas it did not restore thalidomide-induced downregulation of proinflammatory cytokine expression. Importantly, pretreatment with intrathecal injection of the microglial metabolic inhibitor minocycline, IL-10 antibody, ß-endorphin antiserum, and preferred or selective µ-opioid receptor antagonist naloxone or CTAP entirely blocked thalidomide gavage-induced mechanical antiallodynia. Our results demonstrate that thalidomide, but not lenalidomide or pomalidomide, alleviates neuropathic pain, which is mediated by upregulation of spinal microglial IL-10/ß-endorphin expression, rather than downregulation of TNFα expression.
Assuntos
Interleucina-10/biossíntese , Microglia/metabolismo , Neuralgia/tratamento farmacológico , Neuralgia/metabolismo , Talidomida/uso terapêutico , beta-Endorfina/biossíntese , Animais , Células Cultivadas , Relação Dose-Resposta a Droga , Imunossupressores/farmacologia , Imunossupressores/uso terapêutico , Interleucina-10/agonistas , Masculino , Microglia/efeitos dos fármacos , Ratos , Ratos Wistar , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologia , Talidomida/farmacologia , beta-Endorfina/agonistasRESUMO
ß-Endorphin exerts a broad spectrum of physiological activity on mood, immune functions, pain management, reward effects, and behavioral stability. ß-Endorphin is produced in certain neurons within the central and peripheral nervous system but also in the skin, especially in response to ultraviolet radiation. In the present study we have investigated the impact of visible blue light at λâ¯=â¯453â¯nm (BL) on ß-endorphin production of primary human skin keratinocytes (hKC) in-vitro as well as on systemic ß-endorphin formation of whole-body exposed subjects in-vivo. We found that BL irradiation significantly enhanced both keratinocytic ß-endorphin production of hKC cultures as well as systemic ß-endorphin concentrations in light exposed healthy subjects. Interestingly, in hKC cultures elevated ß-endorphin formation was paralleled by significantly increased levels of non-enzymatically generated nitric oxide (NO), whereas elevated systemic ß-endorphin values of BL-exposed subjects were accompanied by enhanced systemic concentration of bioactive NO-derivates. These findings point to a pivotal role of NO in the molecular mechanism of the observed BL-induced effects, and indeed, exogenously applied NO was able to significantly enhance ß-endorphin production in hKC cultures. Thus, our finding of BL-induced increases in systemic ß-endorphin concentration in-vivo can be plausibly explained by an event sequence comprising 1.) BL-driven non-enzymatic formation of NO in the exposed skin tissue, 2.) systemic distribution of cutaneously produced NO in the form of bioactive nitroso compounds, 3.) a subsequent NO-dependent induction of ß-endorphin synthesis in epidermal keratinocytes, and 4.) probably also a NO-dependent modulation of ß-endorphin synthesis in specialized neurons within the central and peripheral nervous system.
Assuntos
Queratinócitos/metabolismo , Óxido Nítrico/química , Pele/metabolismo , beta-Endorfina/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos da radiação , Humanos , Queratinócitos/efeitos da radiação , Luz , Óxido Nítrico/genética , Óxido Nítrico/efeitos da radiação , Pele/crescimento & desenvolvimento , Pele/efeitos da radiação , Raios Ultravioleta/efeitos adversos , beta-Endorfina/biossínteseRESUMO
BACKGROUND AND AIMS: The gut immune, cannabinoid, and opioid systems constitute an integrated network contributing to visceral sensation and pain modulation. We aimed to assess the expression of the µ-opioid receptor (MOR), its ligand ß-endorphin (ß-END), and cannabinoid receptor-2 (CB2 ) in patients with irritable bowel syndrome (IBS) and asymptomatic controls (AC) and their correlation with sex and symptom perception. METHODS: Mucosal biopsies were obtained from the left colon of 31 IBS patients (45% women) with predominant constipation (IBS-C, 9) or diarrhea (IBS-D, 10) or with mixed bowel habits (IBS-M, 12) and 32 AC (44% women) and processed for qRT-PCR, Western blotting, and immunohistochemistry. KEY RESULTS: µ-opioid receptor and CB2 mRNA and protein expression and ß-END protein levels were increased in patients with IBS compared to AC (all Ps=0.021). A significant sex by IBS interaction was found in relation to CB2 mRNA expression (P = .003) with women showing a markedly higher expression to men (P = .035). In contrast, in AC, men had higher expression than women (P = .033). ß-END, MOR, and CB2 immunoreactivities (IR) were localized to CD4+T cells including EMR-1+ eosinophils and CD31+ T cells but not to mast cells. CONCLUSIONS: The increased expression of MOR, ß-END, and CB2 in the mucosa of IBS patients, where they are localized to immune cells, suggests that opioid and cannabinoid systems play an immune-related compensatory role in visceral pain in IBS patients. Further work is necessary to support this hypothesis.
Assuntos
Mucosa Intestinal/metabolismo , Síndrome do Intestino Irritável/metabolismo , Receptor CB2 de Canabinoide/biossíntese , Receptores Opioides mu/biossíntese , beta-Endorfina/biossíntese , Feminino , Humanos , Masculino , Receptor CB2 de Canabinoide/análise , Receptores Opioides mu/análise , Caracteres Sexuais , beta-Endorfina/análiseRESUMO
PURPOSE OF REVIEW: Breast cancer survival has improved motivating the need for better understanding of the sequelae of the disease and its treatments. Lab studies suggest opioids modify cancer cell growth but the association of opioids with cancer progression in humans is not clear. This review aims to summarize recent findings related to opioid use and breast cancer progression. RECENT FINDINGS: Opioid-sparing analgesia may be associated with better survival in cancer patients. In-vitro research suggests that treatment with µ-opioid receptor antagonists inhibits cancer proliferation, and shows some promise for attenuating tumor growth in humans, thereby enhancing survival. Prescription use of opioids does not appear to influence the risk of recurrence in patients, though the evidence comes from a single large registry-based observational study. Ongoing clinical trials are comparing opioid-sparing regional anesthesia with general anesthesia for the risk of breast cancer recurrence. SUMMARY: The association of opioids with breast cancer progression is controversial. Further observational studies are needed. There is currently no clear evidence to suggest that opioid use should be avoided in breast cancer patients because of concerns regarding the risk of breast cancer recurrence.
Assuntos
Analgésicos Opioides/farmacologia , Analgésicos Opioides/uso terapêutico , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/patologia , Anestesia/métodos , Linhagem Celular Tumoral , Ensaios Clínicos como Assunto , Progressão da Doença , Relação Dose-Resposta a Droga , Humanos , Imunidade Celular/efeitos dos fármacos , Recidiva Local de Neoplasia/tratamento farmacológico , Neovascularização Patológica/tratamento farmacológico , Polimorfismo Genético , Prognóstico , Receptores de Estrogênio/efeitos dos fármacos , Receptores Opioides mu/antagonistas & inibidores , beta-Endorfina/biossínteseRESUMO
The glucagon-like peptide-1 (GLP-1) receptor agonist exenatide stimulates microglial ß-endorphin expression and subsequently produces neuroprotection and antinociception. This study illustrated an unrecognized autocrine role of IL-10 in mediation of exenatide-induced ß-endorphin expression. Treatment with exenatide in cultured primary spinal microglia concentration dependently stimulated the expression of the M2 microglial markers IL-10, IL-4, Arg 1, and CD206, but not the M1 microglial markers TNF-α, IL-1ß, IL-6, or CD68. Intrathecal exenatide injection also significantly upregulated spinal microglial expression of IL-10, IL-4, Arg 1, and CD206, but not TNF-α, IL-1ß, IL-6, or CD68. Intrathecal injection of exenatide stimulated spinal microglial expression of IL-10 and ß-endorphin in neuropathic rats. Furthermore, treatment with IL-10 (but not IL-4) stimulated ß-endorphin expression in cultured primary microglia, whereas treatment with ß-endorphin failed to increase IL-10 expression. The IL-10-neutralizing antibody entirely blocked exenatide-induced spinal microglial expression of ß-endorphin in vitro and in vivo and fully blocked exenatide mechanical antiallodynia in neuropathic rats. Moreover, specific cAMP/PKA/p38 signal inhibitors and siRNA/p38ß, but not siRNA/p38α, completely blocked exenatide-induced IL-10 expression in cultured primary microglia. Knock-down of IL-10 receptor-α mRNA using siRNA fully inhibited exenatide-induced spinal microglial ß-endorphin expression and mechanical antiallodynia in neuropathy. Exenatide also markedly stimulated phosphorylation of the transcription factor STAT3 in cultured primary microglia and ß-endorphin stimulation was completely inhibited by the specific STAT3 activation inhibitor. These results revealed that IL-10 in microglia mediated ß-endorphin expression after GLP-1 receptor activation through the autocrine cAMP/PKA/p38ß/CREB and subsequent IL-10 receptor/STAT3 signal pathways.SIGNIFICANCE STATEMENT Activation of GLP-1 receptors specifically and simultaneously stimulates the expression of anti-inflammatory cytokines IL-10 and IL-4, as well as the neuroprotective factor ß-endorphin from microglia. GLP-1 receptor agonism induces ß-endorphin expression and antinociception through autocrine release of IL-10. Activation of GLP-1 receptors stimulates IL-10 and ß-endorphin expression subsequently through the Gs-cAMP/PKA/p38ß/CREB and IL-10/IL-10 receptor-α/STAT3 signal transduction pathways.
Assuntos
Comunicação Autócrina/fisiologia , Receptor do Peptídeo Semelhante ao Glucagon 1/biossíntese , Interleucina-10/biossíntese , Microglia/metabolismo , Medula Espinal/metabolismo , beta-Endorfina/biossíntese , Animais , Animais Recém-Nascidos , Comunicação Autócrina/efeitos dos fármacos , Células Cultivadas , Relação Dose-Resposta a Droga , Exenatida , Expressão Gênica , Receptor do Peptídeo Semelhante ao Glucagon 1/agonistas , Receptor do Peptídeo Semelhante ao Glucagon 1/genética , Interleucina-10/genética , Interleucina-10/farmacologia , Masculino , Microglia/efeitos dos fármacos , Peptídeos/farmacologia , Ratos , Ratos Wistar , Medula Espinal/citologia , Medula Espinal/efeitos dos fármacos , Peçonhas/farmacologia , beta-Endorfina/genéticaRESUMO
The back skin of C57BL/6 mice was exposed to a single 400 mJ cm-2 dose of ultraviolet B (UVB), and parameters of hypothalamic-pituitary-adrenal (HPA) axis in relation to immune activity were tested after 30-90 min following irradiation. Levels of brain and/or plasma corticotropin-releasing hormone (CRH), ß-endorphin, ACTH and corticosterone (CORT) were enhanced by UVB. Hypophysectomy had no effect on UVB-induced increases of CORT. Mitogen-induced IFNγ production by splenocytes from UVB-treated mice was inhibited at 30, 90 min and after 24 h. UVB also led to inhibition of IL-10 production indicating an immunosuppressive effect on both Th1 and Th2 cytokines. Conditioned media from splenocytes isolated from UVB-treated animals had no effect on IFNγ production in cultured normal splenocytes; however, IFNγ increased with conditioned media from sham-irradiated animals. Sera from UVB-treated mice suppressed T-cell mitogen-induced IFNγ production as compared to sera from sham-treated mice. IFNγ production was inhibited in splenocytes isolated from UVB-treated animals with intact pituitary, while stimulated in splenocytes from UVB-treated hypophysectomized mice. Thus, cutaneous exposure to UVB rapidly stimulates systemic CRH, ACTH, ß-endorphin and CORT production accompanied by rapid immunosuppressive effects in splenocytes that appear to be independent of the HPA axis.
Assuntos
Sistema Hipotálamo-Hipofisário/efeitos da radiação , Sistema Hipófise-Suprarrenal/efeitos da radiação , Pele/imunologia , Pele/efeitos da radiação , Raios Ultravioleta/efeitos adversos , Hormônio Adrenocorticotrópico/biossíntese , Animais , Corticosterona/biossíntese , Hormônio Liberador da Corticotropina/biossíntese , Meios de Cultivo Condicionados , Feminino , Interferon gama/sangue , Camundongos Endogâmicos C57BL , beta-Endorfina/biossínteseRESUMO
Immune cell-derived beta-endorphin (END) and other opioid peptides elicit potent and clinically relevant inhibition of pain (analgesia) in inflamed tissue by activation of peripheral opioid receptors. Pro-opiomelanocortin (POMC) is the polypeptide precursor of END and is processed by prohormone convertases (PCs). This study aims to decipher the processing of POMC in lymphocyte subsets in a rat model of unilateral painful hindpaw inflammation. Lymphocytes, isolated from popliteal lymph nodes, were separated into B-cells, T-cells, T-helper cells and cytotoxic T-cells using magnetic cell sorting, and were examined by polymerase chain reaction, immunofluorescence and radioimmunoassay. At 2 h of inflammation, POMC exon 2-3 mRNA was mostly expressed in B- but not in T-cells. Prohormone convertase 1 (PC1) mRNA and protein were upregulated in B-cells and T-helper cells. Prohormone convertase 2 (PC2) was expressed in T- and B-cells, both in inflamed and non-inflamed lymph nodes. END was expressed in B- but not in T-cells. We conclude that POMC, its processing enzymes and END are predominantly expressed in B-lymphocytes at 2 h of paw inflammation.
Assuntos
Linfócitos B/metabolismo , Dor/metabolismo , Pró-Opiomelanocortina/biossíntese , RNA Mensageiro/biossíntese , Serina Endopeptidases/biossíntese , beta-Endorfina/biossíntese , Animais , Linfócitos B/enzimologia , Expressão Gênica , Inflamação/genética , Inflamação/metabolismo , Masculino , Dor/genética , Pró-Opiomelanocortina/genética , Pró-Proteína Convertase 1/biossíntese , Pró-Proteína Convertase 1/genética , Pró-Proteína Convertase 2/biossíntese , Pró-Proteína Convertase 2/genética , RNA Mensageiro/genética , Ratos , Ratos Wistar , Serina Endopeptidases/genética , beta-Endorfina/genéticaRESUMO
BACKGROUND: Giant basal cell carcinomas (GBCCs), (BCC ≥ 5 cm), are often painless, destructive tumors resulting from poorly understood patient neglect. OBJECTIVES: To elucidate etiopathogenic factors distinguishing GBCC from basal cell carcinoma (BCC) and identify predictors for disease-specific death (DSD). METHODS: Case-control study examining clinicopathologic and neuroactive factors (ß-endorphin, met-enkephalin, serotonin, adrenocorticotropic hormone, and neurofilament expression) in GBCC and BCC. Systematic literature review to determine DSD predictors. RESULTS: Thirteen GBCCs (11 patients) were compared with 26 BCCs (25 patients). GBCC significantly differed in size, disease duration, and outcomes; patients were significantly more likely to live alone, lack concern, and have alcoholism. GBCC significantly exhibited infiltrative/morpheic phenotypes, perineural invasion, ulceration, and faster growth. All neuromediators were similarly expressed. Adenoid phenotype was significantly more common in GBCC. Adenoid tumors expressed significantly more ß-endorphin (60% vs. 18%, P = 0.01) and serotonin (30% vs. 4%, P = 0.02). In meta-analysis (n ≤ 311: median age 68 years, disease duration 90 months, tumor diameter 8 cm, 18.4% disease-specific mortality), independent DSD predictors included tumor diameter (cm) (hazard ratio (HR): 1.12, P = 0.003), bone invasion (HR: 4.19, P = 0.015), brain invasion (HR: 8.23, P = 0.001), and distant metastases (HR: 14.48, P = 0.000). CONCLUSIONS: GBCC etiopathogenesis is multifactorial (ie, tumor biology, psychosocial factors). BCC production of paracrine neuromediators deserves further study.
Assuntos
Carcinoma Basocelular/patologia , Serotonina/biossíntese , Neoplasias Cutâneas/patologia , beta-Endorfina/biossíntese , Hormônio Adrenocorticotrópico/análise , Hormônio Adrenocorticotrópico/biossíntese , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma Basocelular/metabolismo , Carcinoma Basocelular/psicologia , Estudos de Casos e Controles , Encefalina Metionina/análise , Encefalina Metionina/biossíntese , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Prognóstico , Estudos Retrospectivos , Serotonina/análise , Neoplasias Cutâneas/metabolismo , Neoplasias Cutâneas/psicologia , Adulto Jovem , beta-Endorfina/análiseRESUMO
INTRODUCTION: Testicular function in the sand rodent Psammomys obesus is subjected to seasonal alternations with a trigger of spermatogenesis in winter and a total quiescence which extends from late spring to summer. The aim of this study was to investigate the distribution of b-endorphin in the testis at the period of winter sexual activity and at its summer regression, and assess the effect of 17 ß-estradiol treatment on testicular morphology and b-endorphin expression. MATERIAL AND METHODS: The adult males were grouped into 4 groups (rest group, sexually active group, rest treated with 17ß-estradiol group and controls at sexual rest injected with olive oil, n = 5 in each group). Using anti-serum against b-endorphin, we studied its testicular expression by Western blot and cellular location by immunohistochemical (IHC) method, respectively. RESULTS: We detected by Western blot a peptide of 3.5 kDa molecular weight corresponding to b-endorphin only in sexually resting and control males. The 17ß-estradiol treatment induced a clear reduction in the b-endorphin band expression compared with the latter. These results were confirmed by the IHC analysis since b-endorphin was only observed in the testis at sexual rest and in controls, in majority of seminiferous tubules at the level of germ cells. The intensity of IHC labeling was significantly different between spermatogonia and spermatocytes I or round spermatids which revealed the strongest labeling. The intense immunoreactivity was also located in Leydig cells and highly significantly varied compared to the germ cells. The 17 ß-estradiol treatment decreased significantly the ß-endorphin signal in germ cells but not in Leydig cells. CONCLUSION: The 17ß-estradiol treatment induces a repressive effect on seasonal testicular endorphinergic system in P. obesus and this action targets exclusively the germ cells.
Assuntos
Estradiol/farmacologia , Gerbillinae/metabolismo , Testículo/efeitos dos fármacos , beta-Endorfina/biossíntese , Animais , Células Germinativas/efeitos dos fármacos , Células Germinativas/metabolismo , Células Intersticiais do Testículo/efeitos dos fármacos , Células Intersticiais do Testículo/metabolismo , Masculino , Azeite de Oliva/farmacologia , Estações do Ano , Túbulos Seminíferos/citologia , Túbulos Seminíferos/efeitos dos fármacos , Túbulos Seminíferos/metabolismo , Comportamento Sexual/efeitos dos fármacos , Espermatócitos/efeitos dos fármacos , Espermatócitos/metabolismo , Espermatogênese/efeitos dos fármacos , Espermatogônias/efeitos dos fármacos , Espermatogônias/metabolismo , Testículo/citologia , Testículo/metabolismo , beta-Endorfina/química , beta-Endorfina/imunologiaRESUMO
We previously found that ultraviolet B (UVB) could stimulate the paraventricular nucleus (PVN) with activation the systemic hypothalamic-pituitary- adrenal (HPA) axis. To investigate whether UVB can also stimulate other hypothalamic nuclei, we tested its effect on the proopiomelanocortin (POMC) related signalling system in the arcuate nucleus (ARC) of female C57BL/6 and FVB albino mice. The shaved back skin of the mice was irradiated with either 100 or 400 mJ/cm2 of UVB. After 1, 3, 6 and 12 h, blood and hypothalamus were collected and processed for gene and protein expression, and measurement of α-MSH and ß-endorphin (ß-END) levels. An in situ immunohistochemical examination was performed for melanocortin receptor 4 (MC4R) and POMC-derived α-MSH. The expression of Pomc and MC4R mRNAs was stimulated, whereas that of AgRP was inhibited after exposure to UVB. It was accompanied by an increased number of both α-MSH- and MC4R-immunoreactive neurons in the ARC, and by increased levels of α-MSH and ß-END (both found in the hypothalamus and plasma). This surprising discovery of UVB stimulating the POMC system in the ARC, accompanied by the increased plasma levels of α-MSH and ß-END, paves the way for exciting areas of research on the communication between the skin and the brain, as well as is suggesting a new role for UVB in regulation of body metabolism.
Assuntos
Núcleo Arqueado do Hipotálamo/metabolismo , Pró-Opiomelanocortina/biossíntese , Pele/efeitos da radiação , Raios Ultravioleta , Proteína Relacionada com Agouti/biossíntese , Proteína Relacionada com Agouti/genética , Animais , Metabolismo Basal , Feminino , Regulação da Expressão Gênica/efeitos da radiação , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos , Pró-Opiomelanocortina/genética , RNA Mensageiro/biossíntese , Receptor Tipo 4 de Melanocortina/biossíntese , Receptor Tipo 4 de Melanocortina/genética , Transdução de Sinais/fisiologia , Organismos Livres de Patógenos Específicos , alfa-MSH/biossíntese , alfa-MSH/sangue , alfa-MSH/genética , beta-Endorfina/biossíntese , beta-Endorfina/sangue , beta-Endorfina/genéticaRESUMO
BACKGROUND: Kahweol is a diterpene present in the oil derived from coffee beans. Although several pharmacological activities of kahweol are already well described in the literature, no study was done in order to assess the analgesic activity of this substance. Thus, the aim of this study was to investigate the possible peripheral antinociceptive effect of kahweol. Considering that the opioid peptides have been implicated in peripheral antinociception induced by non-opioidergic compounds, the present study also evaluated the endogenous opioids involvement in this effect. METHODS: The rat paw pressure test was used, and hyperalgesia was induced by intraplantar injection of prostaglandin E2 (2µg/paw). All drugs were administered subcutaneously in the hindpaws of male Wistar rats. The expression of ß-endorphin was examined by immunohistochemistry in the skin tissue samples of the plantar surface of rat right hindpaws. RESULTS: Intraplantar injection of kahweol (40 and 80µg) induced significant peripheral antinociception. The antinociceptive effect of kahweol was due to a local peripheral action because the higher dose (80µg/paw) did not produce any effect in the contralateral paw. The opioid receptor antagonist naloxone (50 and 100µg/paw) prevented action of kahweol (80µg/paw) and the aminopeptidases inhibitor bestatin (400µg/paw) potentiated the antinociceptive effect of kahweol (40µg/paw). Furthermore, kahweol treatment increased the intensity of ß-endorphin immunoreactivity in the epithelium of rat paws. CONCLUSIONS: The results discussed here provide evidence that kahweol treatment has peripheral antinociceptive effect and suggest that this effect is mediated by the release of endogenous opioids.
Assuntos
Analgésicos/farmacologia , Café/química , Diterpenos/farmacologia , Peptídeos Opioides/farmacologia , Animais , Dinoprostona , Hiperalgesia/induzido quimicamente , Hiperalgesia/tratamento farmacológico , Leucina/análogos & derivados , Leucina/farmacologia , Masculino , Naloxona/farmacologia , Antagonistas de Entorpecentes/farmacologia , Dor/induzido quimicamente , Dor/tratamento farmacológico , Medição da Dor , Peptídeos/farmacologia , Pressão , Ratos , Ratos Wistar , Pele/efeitos dos fármacos , Pele/metabolismo , beta-Endorfina/biossínteseRESUMO
Mammalian skin incorporates a local equivalent of the hypothalamic-pituitary-adrenal (HPA) axis that is critical in coordinating homeostatic responses against external noxious stimuli. Ultraviolet radiation B (UVB) is a skin-specific stressor that can activate this cutaneous HPA axis. Since C57BL/6 (B6) and DBA/2J (D2) strains of mice have different predispositions to sensorineural pathway activation, we quantified expression of HPA axis components at the gene and protein levels in skin incubated ex vivo after UVB or sham irradiation. Urocortin mRNA was up-regulated after all doses of UVB with a maximum level at 50 mJ/cm(2) after 12h for D2 and at 200 mJ/cm(2) after 24h for B6. Proopiomelanocortin mRNA was enhanced after 6h with the peak after 12h and at 200 mJ/cm(2) for both genotypes of mice. ACTH levels in tissue and media increased after 24h in B6 but not in D2. UVB stimulated ß-endorphin expression was higher in D2 than in B6. Melanocortin receptor 2 mRNA was stimulated by UVB in a dose-dependent manner, with a peak at 200 mJ/cm(2) after 12h for both strains. The expression of Cyp11a1 mRNA - a key mitochondrial P450 enzyme in steroidogenesis, was stimulated at all doses of UVB irradiation, with the most pronounced effect after 12-24h. UVB radiation caused, independently of genotype, a dose-dependent increase in corticosterone production in the skin, mainly after 24h of histoculture. Thus, basal and UVB stimulated expression of the cutaneous HPA axis differs as a function of genotype: D2 responds to UVB earlier and with higher amplitude than B6, while B6 shows prolonged (up to 48 h) stress response to a noxious stimulus such as UVB.
Assuntos
Regulação da Expressão Gênica/efeitos da radiação , Sistema Hipotálamo-Hipofisário/metabolismo , Sistema Hipófise-Suprarrenal/metabolismo , Raios Ultravioleta , Hormônio Adrenocorticotrópico/biossíntese , Animais , Enzima de Clivagem da Cadeia Lateral do Colesterol/biossíntese , Relação Dose-Resposta à Radiação , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos DBA , Pró-Opiomelanocortina/biossíntese , Receptor Tipo 2 de Melanocortina/biossíntese , Urocortinas/biossíntese , beta-Endorfina/biossínteseRESUMO
Although peroxisomes exert essential biological functions, cell type-specific features of this important organelle are still only superficially characterized. An intriguing new aspect of peroxisomal function was recently uncovered by the observation that the peptide hormones ß-lipotropin (ß-LPH) and ß-endorphin are localized to peroxisomes in various human tissues. This suggests a functional link between peptide hormone metabolism and peroxisomes. In addition, because endocrine manifestations that affect steroid hormones are often found in patients suffering from inherited peroxisomal disorders, the question has been raised whether peroxisomes are also involved in steroidogenesis. With this chapter, we will review several crucial aspects concerning peroxisomes and hormone metabolism.
Assuntos
Corticosteroides/biossíntese , Córtex Suprarrenal/metabolismo , Hormônios Esteroides Gonadais/biossíntese , Gônadas/metabolismo , Hormônios Peptídicos/biossíntese , Peroxissomos/metabolismo , Animais , Ácidos e Sais Biliares/biossíntese , Humanos , beta-Endorfina/biossíntese , beta-Lipotropina/biossínteseRESUMO
Body and mind interact extensively with each other to control health. Emerging evidence suggests that chronic neurobehavioral stress can promote various tumor growth and progression. The biological reaction to stress involves a chemical cascade initiated within the central nervous system and extends to the periphery, encompassing the immune, endocrine, and autonomic systems. Activation of sympathetic nervous system, such as what happens in the "fight or flight" response, downregulates tumor-suppressive genes, inhibits immune function, and promotes tumor growth. On the other hand, an optimistic attitude or psychological intervention helps cancer patients to survive longer via increase in ß-endorphin neuronal suppression of stress hormone levels and sympathetic outflows and activation of parasympathetic control of tumor suppressor gene and innate immune cells to destroy and clear tumor cells.
Assuntos
Neoplasias da Mama/metabolismo , Imunidade Inata , Proteínas do Tecido Nervoso/metabolismo , Neurônios/metabolismo , Estresse Fisiológico , Estresse Psicológico/metabolismo , beta-Endorfina/metabolismo , Animais , Neoplasias da Mama/etiologia , Neoplasias da Mama/imunologia , Carcinogênese , Proliferação de Células , Citotoxicidade Imunológica , Feminino , Humanos , Glândulas Mamárias Animais/imunologia , Glândulas Mamárias Animais/inervação , Glândulas Mamárias Animais/metabolismo , Glândulas Mamárias Humanas/imunologia , Glândulas Mamárias Humanas/inervação , Glândulas Mamárias Humanas/metabolismo , Proteínas do Tecido Nervoso/biossíntese , Neurônios/imunologia , Estresse Psicológico/fisiopatologia , Transmissão Sináptica , beta-Endorfina/biossínteseRESUMO
BACKGROUND: Prenatal exposure to ethanol (EtOH) reduces the expression of hypothalamic proopiomelanocortin (POMC) gene, known to control various physiological functions including the organismal stress response. In this study, we determined whether the changes in POMC neuronal functions are associated with altered expressions of histone-modifying and DNA-methylating enzymes in POMC-producing neurons, because these enzymes are known to be involved in regulation of gene expression. In addition, we tested whether gestational choline supplementation prevents the adverse effects of EtOH on these neurons. METHODS: Pregnant rat dams were fed with alcohol-containing liquid diet or control diet during gestational days 7 and 21 with or without choline, and their male offspring rats were used during the adult period. Using double-immunohistochemistry, real-time reverse transcription polymerase chain reaction (RT-PCR) and methylation-specific RT-PCR, we determined protein and mRNA levels of histone-modifying and DNA-methylating enzymes and the changes in POMC gene methylation and expression in the hypothalamus of adult male offspring rats. Additionally, we measured the basal- and lipopolysaccharide (LPS)-induced corticosterone levels in plasma by enzyme-linked immunosorbent assay. RESULTS: Prenatal EtOH treatment suppressed hypothalamic levels of protein and mRNA of histone activation marks (H3K4me3, Set7/9, acetylated H3K9, phosphorylated H3S10), and increased the repressive marks (H3K9me2, G9a, Setdb1), DNA-methylating enzyme (Dnmt1), and the methyl-CpG-binding protein (MeCP2). The treatment also elevated the level of POMC gene methylation, while it reduced levels of POMC mRNA and ß-EP and elevated corticosterone response to LPS. Gestational choline normalized the EtOH-altered protein and the mRNA levels of H3K4me3, Set7/9, H3K9me2, G9a, Setdb1, Dnmt1, and MeCP2. It also normalizes the changes in POMC gene methylation and gene expression, ß-EP production, and the corticosterone response to LPS. CONCLUSIONS: These data suggest that prenatal EtOH modulates histone and DNA methylation in POMC neurons that may be resulting in hypermethylation of POMC gene and reduction in POMC gene expression. Gestational choline supplementation prevents the adverse effects of EtOH on these neurons.
Assuntos
Colina/administração & dosagem , Metilação de DNA/efeitos dos fármacos , Histonas/antagonistas & inibidores , Efeitos Tardios da Exposição Pré-Natal/prevenção & controle , Pró-Opiomelanocortina/antagonistas & inibidores , beta-Endorfina/antagonistas & inibidores , Animais , Metilação de DNA/fisiologia , Suplementos Nutricionais , Feminino , Histonas/metabolismo , Hipotálamo/efeitos dos fármacos , Hipotálamo/metabolismo , Masculino , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Gravidez , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Pró-Opiomelanocortina/biossíntese , Ratos , beta-Endorfina/biossínteseRESUMO
BACKGROUND: Animals exposed to alcohol during the developmental period develop circadian disturbances and metabolic problems that often persist during their adult period. In order to study whether alcohol and the circadian clock interact to alter metabolic signaling in the hypothalamus, we determined whether postnatal alcohol feeding in mice permanently alters metabolic sensing in the hypothalamus. Furthermore, we evaluated whether the effect of circadian disruption via Period 2 (Per2) gene mutation prevents alcohol's effects on metabolic signaling in the hypothalamus. METHODS: Per2 mutant and wild-type male and female mice of the same genetic background were given a milk formula containing ethanol (EtOH; 11.34% vol/vol) from postnatal day (PD) 2 to 7 and used for gene expression and peptide level determinations in the hypothalamus at PD7 and PD90. RESULTS: We report here that postnatal alcohol feeding reduces the expression of proopiomelanocortin (Pomc) gene and production of ß-endorphin and α-melanocyte stimulating hormone (α-MSH) in the hypothalamus that persists into adulthood. In addition, expressions of metabolic sensing genes in the hypothalamus were also reduced as a consequence of postnatal alcohol exposure. These effects were not sex-specific and were observed in both males and females. Mice carrying a mutation of the Per2 gene did not show any reductions in hypothalamic levels of Pomc and metabolic genes and ß-endorphin and α-MSH peptides following alcohol exposure. CONCLUSIONS: These data suggest that early-life exposure to alcohol alters metabolic sensing to the hypothalamus possibly via regulating Per2 gene and/or the cellular circadian clock mechanism.
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Etanol/farmacologia , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Hipotálamo/metabolismo , Proteínas Circadianas Period/genética , Fatores Etários , Animais , Animais Recém-Nascidos , Feminino , Regulação da Expressão Gênica no Desenvolvimento/genética , Hipotálamo/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Mutação , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo , Pró-Opiomelanocortina/biossíntese , Fator de Transcrição STAT3/biossíntese , Sirtuína 1/biossíntese , Proteínas Supressoras da Sinalização de Citocina/biossíntese , Fatores de Transcrição/biossíntese , alfa-MSH/biossíntese , beta-Endorfina/biossínteseRESUMO
The experiments show that hypoxia training leads to an increase of the immunoreactivity to corticotropin and beta-endorphin and to an increase of the content of these neuropeptides. In this case, the ratio of the area of the immunoreactive material to the corticotropin/beta-endorphin, and the coefficient of their content in the PVN did not change significantly compared with the control. Thus, the selected hypoxia mode leads to adaptation, which is reflected in the balanced activity of stress-realizing and stress-limiting systems of rats' hypothalamus.
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Adaptação Fisiológica , Hormônio Liberador da Corticotropina/biossíntese , Hipóxia/fisiopatologia , Neurônios/fisiologia , Núcleo Hipotalâmico Paraventricular/fisiopatologia , beta-Endorfina/biossíntese , Adaptação Fisiológica/imunologia , Animais , Hipóxia/imunologia , Hipóxia/metabolismo , Masculino , Neurônios/imunologia , Neurônios/metabolismo , Núcleo Hipotalâmico Paraventricular/imunologia , Núcleo Hipotalâmico Paraventricular/metabolismo , Ratos , Ratos WistarRESUMO
Primary exposure of mice to the nematode Heligmosomoides polygyrus infection reduces inflammation in an experimental model of colitis. The aim of the present investigation was to evaluate whether the reduced inflammation provoked by H. polygyrus L4 larvae in BALB/c mice treated with dextran sulphate sodium is associated with changed expression of opioids in the small intestine and colon. Colitis was induced by 5% Dextran sulphate sodium (DSS) oral administration for 3 days before oral infection with 200 infective larvae (L3) H. polygyrus until the end of the experiment, 6 days post-infection. Clinical disease symptoms were monitored daily. The expressions of proopiomelanocortin POMC1, MOR1 (Oprm1) - opioid receptor and ß-endorphin were determined by RT-PCR, Western blot and immunoassay, respectively, in the colon and small intestine of mice. RT-PCR analysis of colon tissues showed up-regulation of the expression of POMC and MOR1 opioid-dependent genes in mice with DSS-induced colitis. H. polygyrus L4 larvae inhibited DSS-induced colitis symptoms that were correlated with increased IL-1ß, TNF-α, IL-6, myeloperoxidase (MPO) concentration, macrophages infiltration and MOR1, POMC and ß-endorphin increased expression in the small intestine and inhibition of those in the colon.
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Colite/prevenção & controle , Intestinos/fisiologia , Nematospiroides dubius/imunologia , Pró-Opiomelanocortina/biossíntese , Receptores Opioides/biossíntese , beta-Endorfina/biossíntese , Animais , Western Blotting , Colite/induzido quimicamente , Colite/patologia , Citocinas/metabolismo , Sulfato de Dextrana , Perfilação da Expressão Gênica , Larva/imunologia , Macrófagos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Neutrófilos/imunologia , Peroxidase/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Índice de Gravidade de DoençaRESUMO
Periaqueductal gray (PAG) plays a very important role in pain modulation through endogenous opiate peptides including leucine-enkephalin (L-Ek), methionine-enkephalin (M-Ek), ß-endorphin (ß-Ep) and dynorphin A(1-13) (DynA(1-13)). Our pervious study has demonstrated that intra-PAG injection of oxytocin (OXT) increases the pain threshold, and local administration of OXT receptor antagonist decreases the pain threshold, in which the antinociceptive role of OXT can be reversed by pre-PAG administration of OXT receptor antagonist. The experiment was designed to investigate the effect of OXT on endogenous opiate peptides in the rat PAG during the pain process. The results showed that (1) the concentrations of OXT, L-Ek, M-Ek and ß-Ep, not DynA(1-13) in the PAG perfusion liquid were increased after the pain stimulation; (2) the concentrations of L-Ek, M-Ek and ß-Ep, not DynA(1-13) in the PAG perfusion liquid were decreased by the OXT receptor antagonist; (3) the increased pain threshold induced by the OXT was attenuated by naloxone, an opiate receptor antagonist; and (4) the concentrations of L-Ek, M-Ek and ß-Ep, not DynA(1-13) in the PAG perfusion liquid were increased by exogenous OXT administration. The data suggested that OXT in the PAG could influence the L-Ek, M-Ek and ß-Ep rather than DynA(1-13) to participate in pain modulation, i.e. OXT in the PAG participate in pain modulation by influencing the L-Ek, M-Ek and ß-Ep rather than DynA(1-13).
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Microinjeções/métodos , Naloxona/farmacologia , Antagonistas de Entorpecentes/farmacologia , Ocitocina/farmacologia , Limiar da Dor/efeitos dos fármacos , Substância Cinzenta Periaquedutal , Animais , Cateterismo , Dinorfinas/análise , Dinorfinas/biossíntese , Encefalina Leucina/análise , Encefalina Leucina/biossíntese , Encefalina Metionina/análise , Encefalina Metionina/biossíntese , Dor , Medição da Dor , Limiar da Dor/fisiologia , Fragmentos de Peptídeos/análise , Fragmentos de Peptídeos/biossíntese , Substância Cinzenta Periaquedutal/efeitos dos fármacos , Substância Cinzenta Periaquedutal/metabolismo , Radioimunoensaio , Ratos , Ratos Sprague-Dawley , beta-Endorfina/análise , beta-Endorfina/biossínteseRESUMO
Evidence showing expression of endogenous opioids in the mammalian retina is sparse. In the present study we examined a transgenic mouse line expressing an obligate dimerized form of Discosoma red fluorescent protein (DsRed) under the control of the pro-opiomelanocortin promoter and distal upstream regulatory elements to assess whether pro-opiomelanocortin peptide (POMC), and its opioid cleavage product, beta-endorphin, are expressed in the mouse retina. Using double label immunohistochemistry we found that DsRed fluorescence was restricted to a subset of GAD-67-positive cholinergic amacrine cells of both orthotopic and displaced subtypes. About 50% of cholinergic amacrine cells colocalized DsRed and a large fraction of DsRed-expressing amacrine cells was positive for beta-endorphin immunostaining, whereas beta-endorphin-immunoreactive neurons were absent in retinas of POMC null mice. Our findings contribute to a growing body of evidence demonstrating that opioid peptides are an integral component of vertebrate retinas, including those of mammals.