RESUMO
BACKGROUND: Chronic inflammation in adipose tissue together with obesity induces insulin resistance. Inhibitors of chronic inflammation in adipose tissue can be a potent candidate for the treatment of diabetes; however, only a few compounds have been discovered so far. The objective of this study was to find a novel inhibitor that can suppress the inflammatory response in adipose tissue and to elucidate the intracellular signaling mechanisms of the compound. METHODS: To find the active compounds, we established an assay system to evaluate the inhibition of induced MCP-1 production in adipocyte/macrophage coculture in a plant extract library. The active compound was isolated by performing high-performance liquid chromatography (HPLC) and was determined as 4ß-hydroxywithanolide E (4ßHWE) by nuclear magnetic resonance (NMR) and mass spectroscopy (MS) spectral analyses. The effect of 4ßHWE on inflammation in adipose tissue was assessed with adipocyte culture and db/db mice. RESULTS: During the screening process, Physalis pruinosa calyx extract was found to inhibit production of MCP-1 in coculture strongly. 4ßHWE belongs to the withanolide family of compounds, and it has the strongest MCP-1 production inhibitory effect and lowest toxicity than any other withanolides in coculture. Its anti-inflammatory effect was partially dependent on the attenuation of NF-κB signaling in adipocyte. Moreover, in vivo experiments showed that the oral administration of 4ßHWE to db/db mice resulted in the inhibition of macrophage invasion and cytokine expression in adipose tissue after 2 weeks of treatment; improved the plasma adiponectin, non-esterified fatty acids and MCP-1 concentrations; and increased glucose tolerance after 3 to 4 weeks of treatment. CONCLUSIONS: These results suggest that 4ßHWE has anti-inflammatory effect via inhibition of NF-κB activation in adipocyte. Moreover, the attenuation of inflammation in adipocyte has an effect on the inhibition of macrophage accumulation in obese adipose tissue. Consequently, 4ßHWE improves impaired glucose tolerance. Thus, 4ßHWE is a useful natural anti-inflammatory compound to attenuate progression of diabetes and obesity.
Assuntos
Adipócitos/efeitos dos fármacos , Tecido Adiposo/patologia , Quimiocina CCL2/antagonistas & inibidores , Inflamação/tratamento farmacológico , Macrófagos/efeitos dos fármacos , NF-kappa B/antagonistas & inibidores , NF-kappa B/metabolismo , Physalis/química , Extratos Vegetais/farmacologia , Transdução de Sinais/efeitos dos fármacos , Vitanolídeos/farmacologia , Células 3T3-L1 , Adipócitos/metabolismo , Adipócitos/patologia , Tecido Adiposo/efeitos dos fármacos , Tecido Adiposo/metabolismo , Animais , Quimiocina CCL2/biossíntese , Cromatografia Líquida de Alta Pressão , Técnicas de Cocultura , Glucose/metabolismo , Immunoblotting , Resistência à Insulina , Macrófagos/metabolismo , Masculino , Espectrometria de Massas , Camundongos , Camundongos Endogâmicos NOD/metabolismo , Ressonância Magnética Nuclear Biomolecular , Fitoterapia , Vitanolídeos/isolamento & purificaçãoRESUMO
Gustatory and visceral stimulation from food regulates digestion and nutrient use. Free L-glutamate (Glu) release from digested protein is responsible for umami taste perception in the gut. Moreover, monosodium Glu (MSG) is widely used as a flavor enhancer to add umami taste in various cuisines. Recent studies indicate that dietary Glu sensors and their signal transduction system exist in both gut mucosa and taste cells. Oral Glu sensing has been well studied. In this review, we focus on the role of Glu on digestion and absorption of food. Infusion of Glu into the stomach and intestine increase afferent nerve activity of the gastric and the celiac branches of the vagus nerve, respectively. Luminal Glu also evokes efferent nerve activation of the abdominal vagus nerve branches simultaneously. Additionally, intragastric infusion of Glu activates the insular cortex, limbic system, hypothalamus, nucleus tractus solitaries, and amygdala, as determined by functional magnetic resonance imaging, and is able to induce flavor-preference learning as a result of postingestive effects in rats. These results indicate that Glu signaling via gustatory and visceral pathways plays an important role in the processes of digestion, absorption, metabolism, and other physiological functions via activation of the brain.
Assuntos
Sistema Nervoso Autônomo/fisiologia , Digestão , Ácido Glutâmico/metabolismo , Ratos/fisiologia , Transdução de Sinais , Sus scrofa/fisiologia , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Sacarose Alimentar/metabolismo , Preferências Alimentares , Trato Gastrointestinal/metabolismo , Homeostase , Aprendizagem , Metabolismo dos Lipídeos , Período Pós-PrandialRESUMO
Rodents exhibit aversive behavior toward a diet that lacks at least one of the essential amino acids. We sought to determine whether the particular form of anorexia caused by such diets could be ameliorated by the administration of orexigenic peptides while simultaneously analyzing the neural mechanisms underlying anorexia. Rats were fed a valine-deficient diet, which induced severe anorexia (reducing food consumption by 80%). The severe anorexia was associated with a significant decrease in the cerebrospinal fluid valine concentration and hyper-ghrelinemia. Between 6 and 12 days after initiation of the valine-deficient diet, we injected rats twice daily with valine and/or an orexigenic peptide (ghrelin, neuropeptide Y, or agouti-related protein) either i.p. or i.c.v.. We then measured dietary intake. An i.c.v. valine injection allowed earlier food intake compared with an i.p valine injection and increased the density of c-Fos-positive ependymal cells lining the third ventricle. Whereas an i.c.v. injection of ghrelin or neuropeptide Y increased consumption of the valine-deficient diet, i.p injection of ghrelin or i.c.v. injection of agouti-related protein did not. Following i.c.v. administration of either valine or ghrelin, we did not observe complete recovery of consumption of the valine-deficient diet. This may be due to the ineffectiveness of peripheral ghrelin and central agouti-related protein and/or to conditioned aversion to the valine-deficient diet. Since ghrelin is known to be involved in food anticipatory activities, whether the hyper-ghrelinemia observed in valine-deficient rats play role in foraging behavior other than food intake is the future study to be investigated.
Assuntos
Anorexia/metabolismo , Regulação do Apetite/fisiologia , Apetite/fisiologia , Grelina/metabolismo , Valina/deficiência , Proteína Relacionada com Agouti/metabolismo , Proteína Relacionada com Agouti/farmacologia , Animais , Anorexia/tratamento farmacológico , Anorexia/fisiopatologia , Apetite/efeitos dos fármacos , Regulação do Apetite/efeitos dos fármacos , Proteínas Alimentares/metabolismo , Modelos Animais de Doenças , Epêndima/citologia , Epêndima/metabolismo , Comportamento Exploratório/efeitos dos fármacos , Comportamento Exploratório/fisiologia , Comportamento Alimentar/efeitos dos fármacos , Comportamento Alimentar/fisiologia , Alimentos Formulados , Grelina/farmacologia , Hipotálamo/citologia , Hipotálamo/efeitos dos fármacos , Hipotálamo/metabolismo , Masculino , Neuropeptídeo Y/metabolismo , Neuropeptídeo Y/farmacologia , Ratos , Ratos Wistar , Terceiro Ventrículo/citologia , Terceiro Ventrículo/metabolismo , Valina/líquido cefalorraquidiano , Valina/farmacologiaRESUMO
Denatured syngeneic liver tissue prepared by mechanical procedures was intraperitoneally injected into adult C57BL/6 mice. In parallel with a decrease in the total number of lymphocytes in the liver, spleen, and thymus from days 1-7 after the injection, the proportion of the CD4+NK1.1+CD3(int) subset of these cells (i.e. natural killer T or NKT cells) increased in the liver. Even the absolute number of these NKT cells increased in the liver on days 14 and 21. In response to the injection of denatured liver tissue, tissue damage was induced in the liver, as shown by elevated levels of serum transaminases and hepatocyte degeneration observed by electron microscopy. Sera obtained on days 7 and 14 contained autoantibodies including anti-DNA antibodies. The proportion of CD1d(high)B cells in the liver was found to decrease on days 1-7. In other words, denatured liver tissue stimulated both NKT cells and certain B cells in the liver. These results suggest that liver lymphocytes might contain not only autoreactive T cells (e.g. CD3(int) or NKT cells) but also some B cells (e.g. B-1 cells) which produce autoantibodies and that the denatured tissue had the potential to stimulate these lymphocytes and to evoke an autoimmune-like state.
Assuntos
Autoanticorpos/sangue , Doenças Autoimunes/imunologia , Células Matadoras Naturais/imunologia , Fígado/imunologia , Animais , Antígenos/análise , Antígenos Ly , Antígenos de Superfície , Autoantígenos/administração & dosagem , Autoantígenos/química , Autoantígenos/imunologia , Doenças Autoimunes/patologia , Linfócitos B/imunologia , Complexo CD3/análise , Células Cultivadas , Feminino , Injeções Intraperitoneais , Cinética , Lectinas Tipo C , Fígado/patologia , Fígado/ultraestrutura , Ativação Linfocitária , Contagem de Linfócitos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos MRL lpr , Subfamília B de Receptores Semelhantes a Lectina de Células NK , Desnaturação Proteica , Proteínas/análise , Transaminases/sangueRESUMO
OBJECTIVES: To identify genes which are more highly expressed in the peripheral blood mononuclear cells (PBMC) of patients with spondyloarthropathy (SpA), rheumatoid arthritis (RA) and psoriatic arthritis (PsA), in comparison to normal subjects. METHODS: A 588-gene microarray was used as a screening tool to select a panel of such genes from PBMC of these subjects and of normal subjects. Results were then validated by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The following genes were more highly expressed in arthritis patients than in normal subjects: macrophage differentiation marker MNDA (myeloid nuclear differentiation antigen), MRP8 and MRP14 (migratory inhibitory factor-related proteins); signalling molecules JAK3 (janus kinase 3) and MAP kinase p38 (mitogen-activated protein kinase); receptors TNFR2/p75, C-C-chemokine receptor type 1 (CCR1), C-X-C-chemokine receptor type 4 (CXCR4) and integrin beta1; and the cytokines/chemokines interleukin (IL) 1beta and IL-8. Expression of CXCR4 was unexpectedly high among all arthritis subjects. Using RT-PCR, ELISA and immunohistology, expression of stromal cell-derived factor 1 (SDF-1) was demonstrated in arthritis joints. CONCLUSIONS: The CXCR4/SDF-1 is a potential pro-inflammatory axis for RA, PsA and SpA.
Assuntos
Antígenos de Diferenciação/genética , Leucócitos Mononucleares/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , Espondilite Anquilosante/genética , Adolescente , Adulto , Idoso , Antígenos de Diferenciação/sangue , Artrite Psoriásica/sangue , Artrite Psoriásica/genética , Artrite Reumatoide/sangue , Artrite Reumatoide/genética , Quimiocina CXCL12 , Quimiocinas CXC/sangue , Quimiocinas CXC/genética , DNA/análise , Feminino , Marcadores Genéticos , Humanos , Masculino , Pessoa de Meia-Idade , Receptores CXCR4/sangue , Receptores CXCR4/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Espondilite Anquilosante/sangue , Espondilite Anquilosante/patologia , Membrana Sinovial/metabolismo , Membrana Sinovial/patologiaRESUMO
Natural killer T (NKT) cells are mainly present in the liver and thymus, and the majority of these T cells express either a CD4(+) or a double-negative (DN) CD4(-)8(-) phenotype. In the present study, we examined whether such NKT cells were present in the intestine. NKT cells were rare in all sites of the small intestine, including an intraepithelial site. However, a considerable number of NKT cells were found at an intraepithelial site in the large intestine. This result was confirmed by both immunofluorescence and immunohistochemistry. In contrast to conventional NKT cells, NKT cells in the large intestine were CD8(+) or DN CD4(-)8(-). In the case of conventional NKT cells, their existence is known to depend on non-classical MHC class I-like antigens (i. e. CD1d) but not on classical MHC class I antigens. However, the NKT cells in the large intestine were independent of the presence of both CD1d and classical MHC class I antigens. These results were obtained using knockout mice lacking the corresponding genes and molecules. NKT cells in the large intestine were mainly alpha betaTCR(+) (> 75 %) but did not use an invariant chain of Valpha14Jalpha281, which is preferentially used by conventional NKT cells. These NKT cells did not bias the TCR-Vbeta usage toward Vbeta8. These findings suggest that the large intestine is a site in which unconventional NKT cells carrying the CD8(+) phenotype (or DN CD4(-)8(-)) are abundant and that these cells are independent of MHC and MHC-like antigens.
Assuntos
Antígenos CD8/análise , Intestino Grosso/imunologia , Células Matadoras Naturais/fisiologia , Animais , Antígenos/análise , Antígenos de Diferenciação de Linfócitos B/genética , Antígenos de Superfície , Citocinas/biossíntese , Genes Codificadores da Cadeia beta de Receptores de Linfócitos T , Antígenos de Histocompatibilidade Classe II/genética , Imuno-Histoquímica , Mucosa Intestinal/imunologia , Lectinas Tipo C , Camundongos , Camundongos Endogâmicos C57BL , Subfamília B de Receptores Semelhantes a Lectina de Células NK , Proteínas/análiseRESUMO
Given that there are few natural killer T (NKT) cells in the liver of athymic nude mice and in neonatally thymectomized mice, it is still controversial whether all NKT cells existing in the liver are supplied by the thymus or if some such cells develop in the liver. To determine whether or not NKT cells are consistently supplied from the thymus during adult life, thymectomy was conducted in mice at the age of 8 weeks. Interestingly, the proportion and number of CD4+ NKT cells increased or remained unchanged in the liver after adult thymectomy and this phenomenon continued for up to 6 months after thymectomy. The administration of alpha-galactosylceramide induced severe cytopenia (due to apoptosis) of CD4+ NKT cells in the liver on day 1, but subsequent expansion of these NKT cells occurred in thymectomized mice similar to the case in normal mice. However, in thymectomized mice given lethal irradiation (9.5 Gy) and subsequent bone marrow transfer, the population of CD4+ NKT cells no longer expanded in the liver, although that of CD8+ NKT cells did. These results suggest that thymic CD4+ NKT cells, or their progenitors, may migrate to the liver at a neonatal stage but are not supplied from the thymus in the adult stage under usual conditions. CD8+ NKT cells can be generated in the liver.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Células Matadoras Naturais/imunologia , Fígado/imunologia , Timo/imunologia , Animais , Transplante de Medula Óssea , Contagem de Linfócito CD4 , Linfócitos T CD4-Positivos/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Divisão Celular/imunologia , Galactosilceramidas/farmacologia , Imunofenotipagem , Células Matadoras Naturais/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Baço/imunologia , Irradiação Corporal TotalRESUMO
We have been studying polymorphisms of HLA class I and II genes in East Asians including Buryat in Siberia, Mongolian, Han Chinese, Man Chinese, Korean Chinese, South Korean, and Taiwan indigenous populations in collaboration with many Asian scientists. Regional populations in Japan, Hondo-Japanese, Ryukyuan, and Ainu, were also studied. HLA-A, -B, and -DRB1 gene frequencies were subjected to the correspondence analysis and calculation of DA distances. The correspondence analysis demonstrated several major clusters of human populations in the world. "Mongoloid" populations were highly diversified, in which several clusters such as Northeast Asians, Southeast Asians, Oceanians, and Native Americans were observed. Interestingly, an indigenous population in North Japan, Ainu, was placed relatively close to Native Americans in the correspondence analysis. Distribution of particular HLA-A, -B, -DRB1 alleles and haplotypes was also analyzed in relation to migration and dispersal routes of ancestral populations. A number of alleles and haplotypes showed characteristic patterns of regional distribution. For example, B39-HR5-DQ7 (B*3901-DRB1*1406-DQB1*0301) was shared by Ainu and Native Americans. A24-Cw8-B48 was commonly observed in Taiwan indigenous populations, Maori in New Zealand, Orochon in Northeast China, Inuit, and Tlingit. These findings further support the genetic link between East Asians and Native Americans. We have proposed that various ancestral populations in East Asia, marked by different HLA haplotypes, had migrated and dispersed through multiple routes. Moreover, relatively small genetic distances and the sharing of several HLA haplotypes between Ainu and Native Americans suggest that these populations are descendants of some Upper Paleolithic populations of East Asia.
Assuntos
Povo Asiático/genética , Antígenos HLA/genética , Haplótipos/genética , Indígenas Norte-Americanos/genética , Alelos , Sudeste Asiático , Ásia Oriental , Antígenos HLA-D/genética , Antígenos de Histocompatibilidade Classe I/genética , Humanos , Japão , Modelos Genéticos , Análise Multivariada , Polimorfismo Genético , População Branca/genéticaRESUMO
Whether intermediate TCR (TCRint) cells and natural killer T (NKT or NK1.1+TCRint) cells are extrathymically generated remains controversial. This arises from the fact that there are few of these T cells in athymic nude mice and neonatally thymectomized mice. However, when athymic mice were provided with appropriate microenvironments or stimulation, many TCRint cells (mainly NK1.1-) were found to arise in the liver. NKT cells are known to be positively selected by monomorphic major histocompatibility complex (MHC) -like antigens (e.g. CD1d). This is true even if they are CD4+. In other words, a MHC class I-like antigen is restricted to CD4 antigen. This rule is somewhat different from that seen in conventional T cells (i.e. the restriction of class II with CD4 and that of class I and CD8). In the case of NK1.1-TCRint cells, they were selected by polymorphic MHC antigens, but their MHC restriction to CD4 or CD8 antigen was incomplete. This was revealed by experiments of bone marrow transfer with class I (bm 1) or II (bm 12) disparity. Depending on the disparity, a unique cytokine profile in sera was detected. These results suggest that the development of T lineage lymphocytes and MHC restriction to CD4 and CD8 might have occurred in parallell as a phylogenic event, and that NK1.1- extrathymic T cells (i.e. NK1.1-TCRint) are at an intermediate position between NKT cells and conventional T cells in phylogeny.
Assuntos
Transplante de Medula Óssea/imunologia , Hepatócitos/imunologia , Células Matadoras Naturais/imunologia , Complexo Principal de Histocompatibilidade/imunologia , Subpopulações de Linfócitos T/imunologia , Envelhecimento/imunologia , Animais , Complexo CD3/análise , Histocompatibilidade/imunologia , Interferon gama/sangue , Interleucina-12/imunologia , Interleucina-4/sangue , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Nus , MutaçãoRESUMO
When C57Bl/6 mice were exposed to restraint stress for 12 h or 24 h, lymphocytopenia was induced in the liver, spleen, and thymus. We examined which types of lymphocytes were sensitive or resistant to such stress by a immunofluorescence test. T cells of thymic origin were sensitive while NKT and NK cells were resistant. In contrast to the increase in the proportion of NK cells, NK activity of liver lymphocytes against YAC-1 targets decreased at 24 h after stress. On the other hand, their NKT cytotoxicity against syngeneic thymocytes increased in parallel with an increase in their proportion. In perforin -/- B6 mice and B6-gld/gld (Fas ligand-) mice, NK cells were found to mediate cytotoxicity through perforin while NKT cells mediated self-reactive cytotoxicity through Fas ligand. These results suggest that stress increases the proportion of both NK and NKT cells, but that NK cytotoxicity is suppressed while self-reactive NKT cytotoxicity is not, due to a diversity of their functional mechanisms.
Assuntos
Células Matadoras Naturais/imunologia , Subpopulações de Linfócitos/patologia , Linfopenia/etiologia , Estresse Psicológico/imunologia , Animais , Apoptose , Autoimunidade , Citotoxicidade Imunológica/efeitos dos fármacos , Epinefrina/farmacologia , Proteína Ligante Fas , Técnica Indireta de Fluorescência para Anticorpo , Hidrocortisona/farmacologia , Imobilização/efeitos adversos , Fígado/patologia , Subpopulações de Linfócitos/efeitos dos fármacos , Subpopulações de Linfócitos/imunologia , Glicoproteínas de Membrana/deficiência , Glicoproteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Especificidade de Órgãos , Perforina , Proteínas Citotóxicas Formadoras de Poros , Organismos Livres de Patógenos Específicos , Baço/patologia , Estresse Psicológico/complicações , Estresse Psicológico/patologia , Timo/patologiaRESUMO
Beige mice lack natural killer (NK) cytotoxicity, although NK cells are normally present. In recent studies, NK T cells have been newly identified. We therefore examined the number and function of NK T cells in beige mice. The number of NK T cells was at a normal level in the liver of beige mice. NK cytotoxicity was decreased in the liver of these mice, whereas NK T cytotoxicity was intact. When immunochemical staining for perforin was conducted, the majority of NK cells and the minority of NK T cells in beige mice carried a giant granule, containing perforin, in the cytoplasm. In the case of control B6 mice, the majority of NK cells and the minority of NK T cells had multiple, dispersed granules containing perforin. These results suggest that NK T cytotoxicity is unaffected by the beige mutation, owing to their cytotoxicity being mediated without the secretion system of perforin.
Assuntos
Citotoxicidade Imunológica/genética , Células Matadoras Naturais/imunologia , Mutação , Subpopulações de Linfócitos T/imunologia , Animais , Técnicas de Cultura de Células , Citoplasma/química , Citotoxicidade Imunológica/imunologia , Técnicas Imunoenzimáticas , Células Matadoras Naturais/química , Fígado/imunologia , Glicoproteínas de Membrana/análise , Glicoproteínas de Membrana/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Perforina , Proteínas Citotóxicas Formadoras de Poros , Baço/imunologia , Subpopulações de Linfócitos T/químicaRESUMO
c-kit+Lin- cells are present in various immune organs, including the liver, thymus, and bone marrow, where lymphoid, myeloid, or erythroid cells are generated. To compare their properties as lymphoid precursors, c-kit+Lin- cells purified from various organs of B6.Ly5.1 mice were injected into 6.5 Gy-irradiated B6.Ly5.2 mice. Depending on the source of the c-kit cells, the degree of entrance and expansion of lymphoid cells differed in the liver and thymus of recipient mice. c-kit+ cells isolated from the bone marrow entered and expanded prominently in both the liver and thymus, whereas c-kit+ cells from the thymus did not do so at all. On the other hand, c-kit+ cells isolated from the liver and spleen showed an intermediate pattern, namely, they took a long time to enter and expand in the liver and thymus of recipient mice. All of these c-kit+ cells had the potential to give rise to lymphoid cells, which were specific to the liver and thymus, respectively. We previously showed that progenitor cells for extrathymic T cells in the liver and those for conventional T cells in the thymus are not always supplied by the bone marrow, as shown by experiments using parabiosis. Taken together with those previous data, the present results suggest that c-kit+Lin- cells isolated from various immune organs have organ specific properties.
Assuntos
Células da Medula Óssea/citologia , Fígado/citologia , Linfócitos/citologia , Proteínas Proto-Oncogênicas c-kit/análise , Células-Tronco/citologia , Timo/citologia , Animais , Células da Medula Óssea/química , Diferenciação Celular , Subpopulações de Linfócitos , Linfócitos/química , Camundongos , Camundongos Endogâmicos C57BL , Especificidade de Órgãos , Células-Tronco/química , Linfócitos T/química , Linfócitos T/citologiaRESUMO
Sexual dimorphism of the rodent brain is manifested by the epigenetic action of gonadal steroids. Our previous research identified the granulin (grn) precursor gene as a sex steroid-inducible gene, which was shown to be expressed more abundantly in male than female neonates at the mediobasal hypothalamic area. Grn is a 6-kDa polypeptide promoting or inhibiting the growth of epithelial cells and hematocytes in vitro. In this study, effects on male sexual behavior of male were pursued under conditions in which grn gene expression was suppressed during the critical period. To this end, an antisense oligodeoxynucleotide (ODN) of the grn precursor gene was designed, incorporated into inactivated Sendai virus (HVJ)-liposome complexes, and infused into the third ventricle of 2-day-old male rats. Two different control treatments were used: the first consisted of a control sequence ODN that had little homology to known mRNAs; the second of vehicle (HVJ-liposome) alone. After maturation, animals treated with antisense ODN of grn displayed significantly lower scores than control males on various parameters assessing sexual behavior; i.e., mount, intromission, and ejaculation. The antisense ODN, however, did not affect body growth or serum concentrations of testosterone and luteinizing hormone. Further, there was no significant difference in the volume of the sexual dimorphic nucleus of the preoptic area between antisense ODN-treated and control animals. It was shown that inadequate expression of the grn gene in the brain of male neonatal rats during the critical period suppressed the induction of some type of male sexual behavior, suggesting the grn was involved in the process of masculinization of the rat brain.
Assuntos
Peptídeos e Proteínas de Sinalização Intercelular , Oligodesoxirribonucleotídeos Antissenso/administração & dosagem , Precursores de Proteínas/antagonistas & inibidores , Precursores de Proteínas/genética , Comportamento Sexual Animal/efeitos dos fármacos , Animais , Animais Recém-Nascidos , Células Cultivadas , Portadores de Fármacos , Humanos , Hipotálamo/química , Hipotálamo/metabolismo , Injeções Intraventriculares , Lipossomos , Hormônio Luteinizante/sangue , Masculino , Área Pré-Óptica/anatomia & histologia , Área Pré-Óptica/efeitos dos fármacos , Progranulinas , RNA Mensageiro/análise , RNA Mensageiro/genética , Ratos , Ratos Wistar , Respirovirus/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Comportamento Sexual Animal/fisiologia , Testosterona/sangue , Terceiro VentrículoRESUMO
The Ainu people are assumed to be the descendants of pre-agricultural native populations of northern Japan, while the majority of population of present-day Japan (Hondo-Japanese) is considered to have descended mainly from post-neolithic migrants. Sequence-level polymorphisms of the HLA-class I (HLA-A and HLA-B) genes were investigated in DNA samples of 50 Ainu living in Hidaka district, Hokkaido. HLA-A*2402, A*0201, A*0206, A*2601, A*3101, B*1501, B*5101, B*3901, and B*3501 were observed at frequencies of more than 10% and most of these have previously been found in populations of not only Asians but also North and South American Indians. A*68012, which has not so far been detected in Hondo-Japanese, was found in the Ainu (3%). On the other hand, several alleles common in Hondo-Japanese, including HLA-A*3303, A*1101, B*4403, B*5201, B*5401, B*4601, and B*0702 were infrequent in Ainu (0-1%). Correspondence and neighbor-joining analyses of various populations based on HLA-A, -B and -DRB1 gene frequencies enabled distinction between Asian, Native South American, European, and African populations. The Ainu, as well as Tlingit (Na-Dene), were placed midway between other East Asians, including Hondo Japanese, and Native South Americans (Amerindians) in the correspondence analysis. Furthermore, several HLA-A-B and HLA-B-DR-DQ haplotypes common in the Ainu, are shared with some Native American populations. These observations strongly suggest a unique place for the Ainu as descendants of some Upper Paleolithic populations of East Asia, from whom some Native Americans may have descended.
Assuntos
Antígenos HLA-A/genética , Antígenos HLA-B/genética , Alelos , Ásia , Antígenos HLA-A/classificação , Antígenos HLA-B/classificação , Haplótipos , Humanos , JapãoRESUMO
When C57BL/6 mice were partially hepatectomized (PHx), severe lymphocytosis was induced in the liver in the early phase of hepatocyte regeneration (4 to 12 hours after PHx). A major lymphocyte subset expanding in this organ was estimated to be natural killer 1.1(+) (NK1.1(+)) intermediate CD3 (CD3(int)) cells (i.e., NKT cells). CD3(int) cells are extrathymic T cells generated in situ in the liver. These changes were suppressed when mice with PHx were pretreated with a beta-adrenergicD antagonist (i.e., beta-blocker), propranolol (PPL). This might have been caused by sympathetic nerve stimulation during hepatocyte regeneration. An alpha-blocker showed a similar effect, although the magnitude of suppression was lower than that of the beta-blocker. We previously showed that NK and NKT cells express surface beta-adrenergic receptors and are activated in number by sympathetic nerve stimulation. In the present study, NK cytotoxicity mediated by liver lymphocytes obtained from mice with PHx decreased, whereas NKT cytotoxicity against syngeneic thymocytes increased. Purified CD3(int) cells were also found to be able to mediate NKT cytotoxicity against regenerating hepatocytes. These results suggest that sympathetic nerve stimulation after PHx results in subsequent activation of NKT cells and that these NKT cells might be associated with immunologic surveillance during hepatocyte regeneration.
Assuntos
Hepatectomia , Células Matadoras Naturais/imunologia , Regeneração Hepática , Fígado/inervação , Contagem de Linfócitos , Sistema Nervoso Simpático/fisiologia , Antagonistas Adrenérgicos alfa/farmacologia , Antagonistas Adrenérgicos beta/farmacologia , Animais , Complexo CD3/análise , Linfócitos T CD4-Positivos , Citotoxicidade Imunológica , Fígado/imunologia , Subpopulações de Linfócitos/química , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos C57BL , Tamanho do Órgão , Propranolol/farmacologia , Receptores Adrenérgicos beta/análiseRESUMO
When mice were exposed to restraint stress for 12 or 24 h, severe lymphopenia was induced in all immune system organs, including the liver and the thymus. However, in adrenalectomized mice, this response was completely absent. Phenotypic characterization revealed that interleukin (IL)-2Rbeta+CD3int cells (i.e. extrathymic T cells) with CD4+ phenotype and the NK1.1+ subset of CD3int cells (i.e. NKT cells) in the liver as well as the mature conventional T cells in the thymus were resistant to such stress. In adrenalectomized mice, there was no significant change in the distribution of lymphocyte subsets in all tested organs before stress. Interestingly, the number of lymphocytes in the liver and spleen and the proportion of NKT cells in the liver rather increased after stress in these adrenalectomized mice. Therefore, endogenous steroid hormones were indicated to be important in the induction of immunosuppressive states after stress. Among stress associated cytokines, the secretion of tumour necrosis factor (TNF)-alpha was completely suppressed while that of IL-6 was partially suppressed in adrenalectomized mice. These results suggest that endogenous steroid hormones are important for the induction of the stress associated immunosuppression and that NKT cells are resistant to stress, namely, resistant to exposure to endogenous steroid hormones.
Assuntos
Glucocorticoides/fisiologia , Terapia de Imunossupressão , Fígado/imunologia , Baço/imunologia , Estresse Fisiológico/imunologia , Timo/imunologia , Adrenalectomia , Animais , Complexo CD3/biossíntese , Catecolaminas/sangue , Corticosterona/sangue , Citoplasma/imunologia , Citoplasma/metabolismo , Glucocorticoides/sangue , Imunidade Inata , Imunofenotipagem , Interleucina-6/sangue , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Fígado/citologia , Subpopulações de Linfócitos/imunologia , Subpopulações de Linfócitos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Restrição Física , Baço/citologia , Linfócitos T/imunologia , Linfócitos T/metabolismo , Timo/citologia , Fatores de Tempo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Parabiotic pairs of B6.Ly5.1 and B6.Ly5.2 mice were used to investigate how lymphocytes in various organs and various lymphocyte subsets mixed with partner cells. The origin of partner cells was determined by using anti-Ly5.1 mAb in conjunction with immunofluorescence tests. Parabiosis was also produced after the irradiation of B6.Ly5.2 mice at various doses to prepare an immunosuppressive partner. Irrespective of irradiation, lymphocytes and other hematopoietic cells in the bone marrow and lymphocytes in the thymus showed a low mixture of partner cells in comparison with those of all other organs tested. On the other hand, lymphocytes in the blood, spleen, and lymph nodes became a half-and-half mixture of their own cells and partner cells by 14 days after parabiosis. Among lymphocyte subsets, intermediate CD3 cells (i.e., CD3int cells) and NKT cells (i.e., NK1.1+ subset of CD3int cells) in the liver also showed a low mixture of partner cells. The present results raise the possibility that lymphocytes in the bone marrow and thymus, and extrathymic T cells in the liver might be in situ generated from their own preexisting precursor cells. Another observation was that, after irradiation, partner cells showed accelerated mixture even if they showed a low mixture under non-irradiated conditions. However, only lymphocyte subsets with the same phenotype as those of preexisting cells entered the corresponding sites.
Assuntos
Células da Medula Óssea/fisiologia , Células-Tronco Hematopoéticas/fisiologia , Fígado/imunologia , Subpopulações de Linfócitos/fisiologia , Timo/imunologia , Animais , Imunofluorescência , Leucócitos Mononucleares/fisiologia , Fígado/citologia , Linfonodos/citologia , Linfonodos/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Parabiose , Baço/citologia , Baço/imunologia , Linfócitos T/fisiologia , Timo/citologiaRESUMO
NK and extrathymic T cells are abundant in the decidua of the pregnant uterus. To determine how this unique pattern is induced, overall populations of leukocytes were examined in the blood and other tissues in pregnant women. Time-kinetic studies showed that a basal change of leukocytes during pregnancy was granulocytosis and lymphocytopenia in the blood. This change might be due to sympathetic nerve activation during pregnancy, because the administration of catecholamine is known to activate myelopoiesis in the bone marrow. In addition to the numerical change, the functional activation of NK and extrathymic T cells also seemed to be present. This might be due to NK cells and extrathymic T cells (as well as granulocytes), which carry a high density of surface adrenergic receptors. Such functional activation of NK and extrathymic T cells was more prominent in the blood and urine in patients with preeclampsia and hyperemesis gravidarum than in normal pregnant women. The present results suggest that the activation of granulocytes, NK cells, and extrathymic T cells is essential for the maintenance of pregnancy but that overactivation thereof may be responsible for the onset of pregnancy disorders.
Assuntos
Neuroimunomodulação , Gravidez/imunologia , Sistema Nervoso Simpático/fisiologia , Adulto , Feminino , Granulócitos/imunologia , Humanos , Imunofenotipagem , Células Matadoras Naturais/imunologia , Cinética , Leucócitos Mononucleares/imunologia , Subpopulações de Linfócitos , Linfopenia/imunologia , Pré-Eclâmpsia/imunologia , Pré-Eclâmpsia/urina , Receptores Adrenérgicos beta/biossíntese , Linfócitos T/citologia , Linfócitos T/imunologia , Timo/citologia , Fatores de TempoRESUMO
We previously reported that extrathymic T cells (intermediate T-cell receptor cells [TCR(int) cells]) are in situ generated in the parenchymal space of the liver in mice. They subsequently migrate to the sinusoidal lumen. In this study, we characterized how such extrathymic T cells, natural killer (NK) cells, and thymus-derived T cells (high T-cell receptor cells [TCR(high) cells]) localized in the parenchymal space or the sinusoidal lumen of mice. To this end, liver irrigation with physiological saline from the portal vein was performed and the distribution of lymphocyte subsets was compared between the liver (i.e., lymphocytes in the parenchymal space) and the irrigation solution (i.e., lymphocytes in the sinusoidal lumen). Extrathymic T cells and NK cells were found to be abundant in both the liver and sinusoidal lumen. As expected, thymus-derived T cells were abundant in the sinusoidal lumen. However, a significant proportion of thymus-derived T cells were always present in the parenchymal space, even after intensive irrigation with or without collagenase. These results suggest that thymus-derived T cells may consistently infiltrate the parenchymal space from the sinusoidal lumen in normal mice. This possibility was confirmed by (1) the injection of B6 splenic cells (TCR(high) cells) or the thymus graft into B6-nu/nu mice (presence of only TCR(int) cells) and by (2) using parabiotic mice of B6.Ly5.1 and B6.Ly5.2 strains (sharing circulation) in conjunction with immunofluorescence tests and immunohistochemical staining. In other words, inverted routes of migration and homing between extrathymic T cells and thymus-derived T cells exist in the liver.
Assuntos
Fígado/citologia , Linfócitos T/fisiologia , Timo/citologia , Animais , Antígenos/análise , Antígenos de Superfície , Complexo CD3/análise , Movimento Celular , Lectinas Tipo C , Transfusão de Linfócitos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Nus , Subfamília B de Receptores Semelhantes a Lectina de Células NK , Proteínas/análise , Receptores de Interleucina-2/análise , Irrigação TerapêuticaRESUMO
The prolactin (PRL) surge in cycling rats during the proestrous afternoon is an inducer of apoptotic cell death in luteal cells. This luteolytic action of PRL is peculiar, because PRL may be categorized as a survival factor, if other known physiological functions of PRL are taken into account. Here we analyzed the underlying molecular/cellular mechanisms of this PRL-induced apoptosis. Corpora lutea (CL) were prepared from the ovary on the proestrous day and cultured with or without PRL (2 microg/ml). An addition of PRL to the culture medium induced DNA breakdown in the nuclei of cells mostly identified as steroidogenic by 3beta-HSD activity staining, and the number of 3beta-HSD-positive cells were significantly decreased, indicating the induction of apoptotic cell death by PRL among luteal cells in culture. Next, the expression of membrane form-Fas ligand (mFasL) in the luteal cell lysate was quantified, because Fas receptor is known to have an exact physiological role in luteolysis. An addition of PRL increased the expression of mFasL. Immunostaining and TUNEL assay on regressing CL revealed that both CD3-positive cells and FasL-positive cells were co-localized in the regions where apoptosis convergently occurred. Moreover, an addition of concanavalin A (ConA), a T-cell specific activator, to the culture mimicked the PRL action by inducing apoptosis in luteal cells and enhancing the expression of mFasL. These data suggest that the CD3-positive T lymphocyte in the CL is at least one of the PRL-effector cell species during the process of luteolysis in rats, and that FasL expression of these cells is upregulated by PRL.