RESUMEN
Lipid mediators are known to play crucial roles not only in the onset of the inflammatory response but also in the induction of resolution of inflammation. Here, we report that palmitoylethanolamide (PEA), an endogenous N-acylethanolamine, can suppress the inflammation induced by Toll-like receptor (TLR) signaling both in vitro and in vivo. PEA was found to be significantly reduced in the serum and spleen of lupus-prone MRL/lpr mice analyzed by lipidomics. PEA suppressed pro-inflammatory cytokine production in a mouse macrophage cell line stimulated with TLR ligands such as lipopolysaccharide, peptidoglycan, poly (I:C), imiquimod, and CpG-ODN. PEA also inhibited both mRNA and protein levels of IL-6 in bone marrow-derived dendritic cells (BMDCs) and B cells stimulated with CpG-ODN. Augmentation of cell surface CD86 and CD40 on BMDCs and B cells, IgM production, and cell proliferation of B cells in response to CpG-ODN were attenuated by PEA. Moreover, PEA treatment significantly reduced mortality and serum IL-6 levels in mice injected with CpG-ODN plus D-galactosamine. Taken together, PEA ameliorates inflammation induced by TLR signaling, which could be a novel therapeutic target for inflammatory disorders.
Asunto(s)
Interleucina-6 , Receptor Toll-Like 9 , Amidas , Animales , Cromatografía Liquida , Etanolaminas , Inflamación/tratamiento farmacológico , Interleucina-6/metabolismo , Lipidómica , Ratones , Ratones Endogámicos MRL lpr , Ácidos Palmíticos , Espectrometría de Masas en Tándem , Receptores Toll-LikeRESUMEN
Antibiotics sometimes exert adverse effects on the pathogenesis of colitis due to the dysbiosis resulting from the disruption of gut homeostasis. However, the precise mechanisms underlying colitogenic effects of antibiotic-induced colitis are largely unknown. Here, we show a novel murine fecal occult bleeding model induced by the combinatorial treatment of ampicillin and vancomycin, which is accompanied by an enlarged cecum, upregulation of pro-inflammatory cytokines IL-6 and IL-12, a reduction in Ki-67-positive epithelial cell number and an increase in the apoptotic cell number in the colon. Moreover, gas chromatography-tandem mass analysis showed that various kinds of metabolites, including glutamic acid and butyric acid, were significantly decreased in the cecal contents. In addition, abundance of butyric acid producer Clostridiales was dramatically reduced in the enlarged cecum. Interestingly, supplementation of monosodium glutamate or its precursor glutamine suppressed colonic IL-6 and IL-12, protected from cell apoptosis and prevented fecal occult blood indicating that the reduced level of glutamic acid is a possible mechanism of antibiotic-induced fecal occult bleeding. Our data showed a novel mechanism of antibiotic-induced fecal occult bleeding providing a new insight into the clinical application of glutamic acid for the treatment of antibiotic-induced colitis.
Asunto(s)
Antibacterianos/administración & dosificación , Antibacterianos/efectos adversos , Colon/patología , Células Epiteliales/patología , Enfermedades Metabólicas/complicaciones , Sangre Oculta , Administración Oral , Ampicilina/administración & dosificación , Ampicilina/farmacología , Animales , Péptidos Catiónicos Antimicrobianos/metabolismo , Ácido Butírico/farmacología , Metabolismo de los Hidratos de Carbono/efectos de los fármacos , Ciego/microbiología , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Neoplasias del Colon/patología , Citocinas/metabolismo , Glutamina/administración & dosificación , Metabolismo de los Lípidos/efectos de los fármacos , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Metaboloma/efectos de los fármacos , Metagenómica , Ratones , Microbiota/efectos de los fármacos , Microbiota/genética , Células RAW 264.7 , Regeneración/efectos de los fármacos , Glutamato de Sodio/administración & dosificación , Especificidad de la Especie , Vancomicina/administración & dosificación , Vancomicina/farmacologíaRESUMEN
Sialadenitis is an inflammatory condition affecting the salivary glands including the parotid, submandibular, and sublingual glands. There are several different types of sialadenitis, each with different sites of predilection. However, the pathogenic mechanism underlying the tissue specificity of sialadenitis is largely unknown. TRAF6 is a cytoplasmic adaptor protein that is necessary for the activation of dendritic cells in response to Toll-like receptor ligands, thereby regulating innate immune responses. We previously demonstrated that T cell-specific TRAF6-deficient mice (TRAF6ΔT mice) spontaneously develop systemic inflammatory disease. Here, we show that salivary secretion is reduced in TRAF6ΔT mice due to sialadenitis that occurs in the parotid and submandibular glands, but not the sublingual glands. Consistent with pathological findings, both CD4+ and CD8+ T cells predominantly infiltrated the submandibular glands; however, sublingual infiltration was rare in TRAF6ΔT mice. The TH1 cytokine IFN-γ, the TH1 cell attractant chemokine CCL2, and its cognate receptor CCR2 were upregulated concomitantly in both the submandibular and sublingual glands. Interestingly, the TH17â¯cell attractant chemokine CCL20 and its cognate receptor CCR6 were selectively increased in the submandibular glands, but not in the sublingual glands of TRAF6ΔT mice. Thus, the expression of TRAF6 in T cells might be implicated in tissue-specific sialadenitis by regulating the chemokine-chemokine receptor system.
Asunto(s)
Enfermedades Autoinmunes/metabolismo , Quimiocinas/metabolismo , Receptores de Quimiocina/metabolismo , Sialadenitis/metabolismo , Linfocitos T/metabolismo , Factor 6 Asociado a Receptor de TNF/genética , Animales , Quimiocina CCL2/metabolismo , Citoplasma/metabolismo , Inflamación , Ratones , Ratones Noqueados , Glándula Parótida/metabolismo , Receptores CCR2/metabolismo , Glándulas Salivales/metabolismo , Sialadenitis/inmunología , Glándula Submandibular/metabolismo , Células TH1/metabolismo , Células Th17/metabolismo , Regulación hacia ArribaRESUMEN
A series of fatty acid amides were synthesized and their peroxisome proliferator-activated receptor α (PPAR-α) agonistic activities were evaluated in a normal rat liver cell line, clone 9. The mRNAs of the PPAR-α downstream genes, carnitine-palmitoyltransferase-1 and mitochondrial 3-hydroxy-3-methylglutaryl-CoA synthase, were determined by real-time reverse transcription-polymerase chain reaction (RT-PCR) as PPAR-α agonistic activities. We prepared nine oleic acid amides. Their PPAR-α agonistic activities were, in decreasing order, N-oleoylhistamine (OLHA), N-oleoylglycine, Oleamide, N-oleoyltyramine, N-oleoylsertonin, and Olvanil. The highest activity was found with OLHA. We prepared and evaluated nine N-acylhistamines (N-acyl-HAs). Of these, OLHA, C16:0-HA, and C18:1Δ(9)-trans-HA showed similar activity. Activity due to the different chain length of the saturated fatty acid peaked at C16:0-HA. The PPAR-α antagonist, GW6471, inhibited the induction of the PPAR-α downstream genes by OLHA and N-oleoylethanolamide (OEA). These data suggest that N-acyl-HAs could be considered new PPAR-α agonists.
Asunto(s)
Amidas/farmacología , Ácidos Grasos/farmacología , PPAR alfa/agonistas , Amidas/síntesis química , Amidas/química , Animales , Línea Celular , Ácidos Grasos/síntesis química , Ácidos Grasos/química , Hígado/efectos de los fármacos , Hígado/metabolismo , Estructura Molecular , PPAR alfa/genética , PPAR alfa/metabolismo , ARN Mensajero/genética , Ratas , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
In this study, we have proposed and demonstrated experimentally a novel monitoring device of single mouse embryo activity after in vitro fertilization (IVF) using a semiconductor-based field effect transistor (FET). The FET biosensor realized to detect it noninvasively, quantitatively, and continuously by change of hydrogen ions with positive charges, which were induced by dissolved carbon dioxide due to cellular respiration activity during cleavage. The electrical signal of FET biosensor should become an effective indication to evaluate objectively single embryo activity as its morphology is observed subjectively after IVF. The platform based on the FET biosensor will contribute to promote elective single embryo transfer (eSET) in human assisted reproductive technology (ART).
Asunto(s)
Técnicas Biosensibles/métodos , Transferencia de un Solo Embrión/métodos , Transistores Electrónicos , Animales , Técnicas Biosensibles/instrumentación , Técnicas de Cultivo de Embriones/instrumentación , Técnicas de Cultivo de Embriones/métodos , Femenino , Masculino , Ratones , Ratones Endogámicos ICR , Embarazo , Transferencia de un Solo Embrión/instrumentaciónRESUMEN
Effective delivery of protein therapeutics into the brain remains challenging because of difficulties associated with crossing the blood-brain barrier (BBB). To overcome this problem, many researchers have focused on antibodies binding the transferrin receptor (TfR), which is expressed in endothelial cells, including those of the BBB, and is involved in receptor-mediated transcytosis (RMT). RMT and anti-TfR antibodies provide a useful means of delivering therapeutics into the brain, but the anti-TfR antibody has a short half-life in blood because of its broad expression throughout the body. As a result, anti-TfR antibodies are only maintained at high concentrations in the brain for a short time. To overcome this problem, we developed a different approach which slows down the export of therapeutic antibodies from the brain by binding them to a brain-specific antigen. Here we report a new technology, named AccumuBrain, that achieves both high antibody concentration in the brain and a long half-life in blood by binding to myelin oligodendrocyte glycoprotein (MOG), which is specifically expressed in oligodendrocytes. We report that, using our technology, anti-MOG antibody levels in the brains of mice (Mus musculus) and rats (Rattus norvegicus) were increased several tens of times for a period of one month. The mechanism of this technology is different from that of RMT technologies like TfR and would constitute a breakthrough for central nervous system disease therapeutics.
Asunto(s)
Anticuerpos Antiidiotipos/inmunología , Barrera Hematoencefálica/inmunología , Glicoproteína Mielina-Oligodendrócito/inmunología , Receptores de Transferrina/inmunología , Animales , Anticuerpos/inmunología , Anticuerpos Antiidiotipos/genética , Barrera Hematoencefálica/efectos de los fármacos , Encéfalo/efectos de los fármacos , Encéfalo/inmunología , Epítopos/inmunología , Humanos , Ratones , Glicoproteína Mielina-Oligodendrócito/genética , Especificidad de Órganos/inmunología , Unión Proteica/inmunología , Ratas , Transducción de Señal/inmunología , Transcitosis/genética , Transcitosis/inmunologíaRESUMEN
BACKGROUND: Both environmental and genetic factors have been implicated in the induction of autoimmune disease. Therefore, it is important to understand the pathophysiological significance of the gut microbiota and host genetic background that contribute to an autoimmune disease such as inflammatory bowel disease (IBD). We have previously reported that mice deficient for suppressor of cytokine signaling-1 (SOCS1), in which SOCS1 expression was restored in T and B cells on an SOCS1-/- background (SOCS1-/-Tg mice), developed systemic autoimmune diseases accompanied by spontaneous colitis. METHODS: To investigate whether the proinflammatory genetic background affects the gut microbiota, we used SOCS1-/-Tg mice as a model of spontaneous chronic colitis. Fecal samples were collected from SOCS1-/-Tg mice and SOCS1+/+Tg (control) mice at 1 and 6 months of age, and the fecal bacterial 16S ribosomal RNA genes were sequenced using the Illumina MiSeq platform. RESULTS: Gut microbial diversity was significantly reduced and the intestinal bacterial community composition changed in SOCS1-/-Tg mice in comparison with the control mice. Interestingly, the population of Prevotella species, which is known to be elevated in ulcerative colitis and colorectal cancer patients, was significantly increased in SOCS1-/-Tg mice regardless of age. CONCLUSION: Taken together, these results suggest that the proinflammatory genetic background owing to SOCS1 deficiency causes dysbiosis of the gut microbiota, which in turn generates a procolitogenic environment.
RESUMEN
Zika fever, a mosquito-borne infectious disease caused by Zika virus (ZIKV), is an epidemic disease for which no effective therapy has been established. The recent outbreaks of ZIKV in Brazil and French Polynesia have been linked to a considerable increase in the incidence of fetal microcephaly and other diseases such as Guillain-Barre syndrome. Because there is currently no specific therapy or vaccine, the early exploitation of a method to prevent expansion of ZIKV is a high priority. To validate commonly used antiviral drugs, we evaluated the effect of ribavirin, a drug used to treat hepatitis C with interferon-ß (IFN-ß), on ZIKV replication. In mammalian cells, we observed an inhibitory effect of ribavirin on ZIKV replication and ZIKV-induced cell death without cytotoxic effect. Furthermore, we found that STAT1-deficient mice, which lack type I IFN signaling, were highly sensitive to ZIKV infection and exhibited lethal outcome. Ribavirin abrogated viremia in ZIKV-infected STAT-1-deficient mice. These data suggest that the inhibition of viral RNA-dependent RNA polymerases may be effective for treatment of ZIKV infection. Our data provide a new insight into the mechanisms for inhibition of ZIKV replication and prevention of Zika fever.
Asunto(s)
Antivirales/farmacología , Ribavirina/farmacología , Factor de Transcripción STAT1/deficiencia , Infección por el Virus Zika/tratamiento farmacológico , Virus Zika/efectos de los fármacos , Animales , Antivirales/administración & dosificación , Antivirales/uso terapéutico , Chlorocebus aethiops , Replicación del ADN/efectos de los fármacos , Ratones , Ribavirina/administración & dosificación , Ribavirina/uso terapéutico , Factor de Transcripción STAT1/genética , Células Vero , Viremia/tratamiento farmacológico , Viremia/virología , Replicación Viral/efectos de los fármacos , Virus Zika/fisiología , Infección por el Virus Zika/virologíaRESUMEN
The therapeutic use of neurotrophic factors to treat neurodegenerative disorders, including Alzheimer's disease, is considered feasible. Magnolol and honokiol, constituents of the Magnolia plant, are small organic compounds with neurotrophic activity. We investigated whether magnolol and honokiol can prevent age-related learning and memory impairment and cholinergic deficits in senescence-accelerated mice (SAM). Magnolol (1, 10 mg/kg) or honokiol (0.1, 1 mg/kg) were orally administered to SAMP8 mice once a day for 14 days in 2-month-old mice. Learning and memory performance were evaluated by passive avoidance tests and location and object novelty recognition tests. SAMP8 mice showed significant impairment of learning and memory at 4 and 6 months of age. This age-related learning and memory impairment was prevented by pretreatment with either magnolol (10 mg/kg) or honokiol (1 mg/kg). Cholinergic neuron densities in the medial septum and vertical limb of the diagonal band of the forebrain were evaluated by an immunohistochemical analysis of choline acetyltransferase (ChAT). SAMP8 mice showed a significant cholinergic deficit at 6 months of age. These age-related cholinergic deficits were prevented by treatment with either magnolol (10 mg/kg) or honokiol (1 mg/kg). Moreover, SAMP8 mice showed decreased activity of Akt, a member of the prosurvival pathway, in the forebrain at 2 months of age. A 14-day treatment with either magnolol (10 mg/kg) or honokiol (1 mg/kg) enhanced phosphorylation of Akt in the forebrain at 2 months of age. These results suggest that magnolol and honokiol prevent age-related learning and memory impairment by preserving cholinergic neurons in the forebrain. These compounds may have potential therapeutic applications to various neurodegenerative disorders.