RESUMEN
Fulminant hepatitis is characterized by rapid and massive immune-mediated liver injury. Dosage-sensitive sex reversal-adrenal hypoplasia congenita critical region on the X chromosome, gene 1 (DAX1; NR0B1) represses the transcription of various genes. Here, we determine whether DAX1 serves as a regulator of inflammatory liver injury induced by concanavalin A (ConA). C57BL/6J (WT), myeloid cell-specific Dax1 knockout (MKO), and hepatocyte-specific Dax1 knockout (LKO) mice received single intravenous administration of ConA. Histopathological changes in liver and plasma alanine aminotransferase and aspartate aminotransferase levels in Dax1 MKO mice were comparable with those in WT mice following ConA administration. Unlike Dax1 MKO mice, Dax1 LKO mice were greatly susceptible to ConA-induced liver injury, which was accompanied by enhanced infiltration of immune cells, particularly CD4+ and CD8+ T cells, in the liver. Factors related to T-cell recruitment, including chemokines and adhesion molecules, significantly increased following enhanced and prolonged phosphorylation of NF-κB p65 in the liver of ConA-administered Dax1 LKO mice. This is the first study to demonstrate that hepatocyte-specific DAX1 deficiency exacerbates inflammatory liver injury via NF-κB p65 activation, thereby causing T-cell infiltration by modulating inflammatory chemokines and adhesion molecules. Our results suggest DAX1 as a therapeutic target for fulminant hepatitis treatment.
Asunto(s)
Linfocitos T CD8-positivos , Necrosis Hepática Masiva , Ratones , Animales , FN-kappa B , Ratones Endogámicos C57BL , Hepatocitos , Transducción de Señal , Concanavalina A/toxicidad , Linfocitos T CD4-PositivosRESUMEN
Acetaminophen (APAP) is a widely used analgesic and antipyretic drug, but its overdose can cause acute liver failure. The dosage-sensitive sex reversal adrenal hypoplasia congenita critical region on the X chromosome, gene 1 (DAX-1, NR0B1), is an orphan nuclear receptor that acts as a transcriptional co-repressor of various genes. In this study, we identified the role of DAX-1 in APAP-induced liver injury using hepatocyte-specific Dax-1 knockout (Dax-1 LKO) mice. Mouse primary hepatocytes were used as a comparative in vitro study. APAP overdose led to decreased plasma alanine aminotransferase and aspartate aminotransferase levels in Dax-1 LKO mice compared to C57BL/6J (WT) controls, accompanied by reduced liver necrosis. The expression of the genes encoding the enzymes catalyzing glutathione (GSH) synthesis and metabolism and antioxidant enzymes was increased in the livers of APAP-treated Dax-1 LKO mice. The rapid recovery of GSH levels in the mitochondrial fraction of APAP-treated Dax-1 LKO mice led to reduced reactive oxygen species levels, resulting in the inhibition of the prolonged JNK activation. The hepatocyte-specific DAX-1 deficiency increased the protein expression of nuclear factor erythroid 2-related factor 2 (Nrf2) compared with WT controls after APAP administration. These results indicate that DAX-1 deficiency in hepatocytes protects against APAP-induced liver injury by Nrf2-regulated antioxidant defense.
Asunto(s)
Antipiréticos , Enfermedad Hepática Inducida por Sustancias y Drogas , Receptor Nuclear Huérfano DAX-1 , Factor 2 Relacionado con NF-E2 , Acetaminofén/toxicidad , Alanina Transaminasa/metabolismo , Animales , Antioxidantes/metabolismo , Aspartato Aminotransferasas/metabolismo , Enfermedad Hepática Inducida por Sustancias y Drogas/genética , Enfermedad Hepática Inducida por Sustancias y Drogas/metabolismo , Enfermedad Hepática Inducida por Sustancias y Drogas/prevención & control , Proteínas Co-Represoras/metabolismo , Receptor Nuclear Huérfano DAX-1/genética , Glutatión/metabolismo , Hepatocitos/metabolismo , Hígado/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Factor 2 Relacionado con NF-E2/genética , Factor 2 Relacionado con NF-E2/metabolismo , Receptores Nucleares Huérfanos/metabolismo , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Fulminant hepatitis is a serious inflammatory condition of the liver characterized by massive necrosis of liver parenchyma following excessive immune cell infiltration into the liver, and possibly causing sudden hepatic failure and medical emergency. However, the underlying mechanisms are not fully understood. Here, we investigated the role of cyclic AMP-responsive element-binding protein, hepatocyte specific (CREBH) in concanavalin A (ConA)-driven hepatitis-evoked liver injury. C57BL/6J (WT) and Crebh knockout (KO) mice injected with ConA (7.5 or 25 mg/kg) and bone marrow (BM) chimeric mice, generated by injection of BM cells into sub-lethally irradiated recipients followed by ConA injection (22.5 or 27.5 mg/kg) 8 weeks later, were used for in vivo study. Primary mouse hepatocytes and HEK293T cells were used for a comparative in vitro study. Crebh KO mice are highly susceptible to ConA-induced liver injury and prone to death due to increased neutrophil infiltration driven by enhanced hepatic expression of neutrophil-attracting chemokines. Notably, BM chimera experiment demonstrated that Crebh-deficient hepatocytes have an enhanced ability of recruiting neutrophils to the liver, thereby promoting hepatotoxicity by ConA. Intriguingly, in vitro assays showed that p65, a subunit of NF-κB and common transcription factor for various chemokines, dependent transactivation was inhibited by CREBH. Furthermore, p65 expression was inversely correlated with CREBH level in ConA-treated mice liver and TNFα-stimulated primary mouse hepatocytes. This is the first demonstration that CREBH deficiency aggravates inflammatory liver injury following chemokine-dependent neutrophil infiltration via NF-κB p65 upregulation. CREBH is suggested to be a novel therapeutic target for treatment of fulminant hepatitis.
Asunto(s)
Quimiocinas/metabolismo , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/genética , Necrosis Hepática Masiva/patología , Infiltración Neutrófila , Factor de Transcripción ReIA/metabolismo , Animales , Enfermedad Hepática Inducida por Sustancias y Drogas/etiología , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Concanavalina A/toxicidad , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Citocinas/sangre , Citocinas/metabolismo , Células HEK293 , Humanos , Masculino , Necrosis Hepática Masiva/inducido químicamente , Necrosis Hepática Masiva/metabolismo , Ratones Endogámicos C57BL , Ratones Noqueados , Infiltración Neutrófila/efectos de los fármacos , Regulación hacia Arriba/efectos de los fármacosRESUMEN
Gastritis and gastric ulcers are common gastric diseases that are caused by infection, drugs, alcohol consumption, or stress. These conditions lead to increased inflammatory cytokines and recruitment of leukocytes, which damage the stomach mucosa and exacerbate disease severity. Sicyos angulatus (SA), an annual vine in the Cucurbitaceae family, is known to have an anti-inflammatory effect, but its efficacy for preventing gastritis and gastric ulcers has not yet been evaluated. In the present study, we investigated the gastroprotective effect of SA using a hydrochloric acid/ethanol-induced gastric mucosal injury mouse model and lipopolysaccharide (LPS)-stimulated KATO III cells. Macroscopic analysis revealed a reduction in gastric ulcer area. Similarly, histopathological analysis showed a dose-dependent decrease in gastric mucosal injury, with significant improvement at 750 mg/kg of SA treatment. Gene expressions of inflammatory cytokines, chemokines, and adhesion molecule were reduced in the SA-administered group. Immunohistochemical staining indicated that SA significantly decreased neutrophil infiltration in the lamina propria and epithelium of the stomach. Kaempferol, a major bioactive flavonoid of SA, also improved gastric injury by reducing macroscopic and microscopic lesions, inflammatory mediator gene expression, and neutrophil infiltration. Furthermore, both SA and kaempferol downregulated LPS-mediated increases in inflammatory cytokines and chemokines following inhibition of p38 and c-Jun N-terminal kinase (JNK) phosphorylation in KATO III cells. These results suggest that SA can ameliorate gastric mucosal injury by inhibiting the recruitment of inflammatory cells, particularly neutrophils, and by suppressing p38 and JNK phosphorylation.
RESUMEN
Akkermansia muciniphila has received great attention because of its beneficial roles in gut health by regulating gut immunity, promoting intestinal epithelial development, and improving barrier integrity. However, A. muciniphila-derived functional molecules regulating gut health are not well understood. Microbiome-secreted proteins act as key arbitrators of host-microbiome crosstalk through interactions with host cells in the gut and are important for understanding host-microbiome relationships. Herein, we report the biological function of Amuc_1409, a previously uncharacterised A. muciniphila-secreted protein. Amuc_1409 increased intestinal stem cell (ISC) proliferation and regeneration in ex vivo intestinal organoids and in vivo models of radiation- or chemotherapeutic drug-induced intestinal injury and natural aging with male mice. Mechanistically, Amuc_1409 promoted E-cadherin/ß-catenin complex dissociation via interaction with E-cadherin, resulting in the activation of Wnt/ß-catenin signaling. Our results demonstrate that Amuc_1409 plays a crucial role in intestinal homeostasis by regulating ISC activity in an E-cadherin-dependent manner and is a promising biomolecule for improving and maintaining gut health.
Asunto(s)
Verrucomicrobia , beta Catenina , Masculino , Ratones , Animales , beta Catenina/metabolismo , Verrucomicrobia/metabolismo , Intestinos , Cadherinas/metabolismo , AkkermansiaRESUMEN
BACKGROUND: Neuroinflammation plays an important role in cognitive decline and memory impairment in neurodegenerative disorders. Previously, we demonstrated that Humulus japonicus (HJ) has anti-inflammatory effects in rodent models of Alzheimer's disease and Parkinson's disease. The present study aimed to examine the protective potential of HJ extracts against lipopolysaccharide (LPS)-induced cognitive impairment and scopolamine-induced amnesia in mouse models. Cognitive improvement of mice was investigated by novel object recognition test. For analyzing effects on neuroinflammation, immunohistochemistry and quantitative real-time polymerase chain reaction (qRT-PCR) assays were performed. RESULTS: We found that the oral administration of HJ significantly improved cognitive dysfunction induced by LPS in a novel object recognition test. The LPS-induced activation of microglia was notably decreased by HJ treatment in the cortex and hippocampus. HJ administration with LPS also significantly increased the mRNA expression of interleukin (IL)-10 and decreased the mRNA expression of IL-12 in the parietal cortex of mice. The increased expression of LPS-induced complement C1q B chain (C1bq) and triggering receptor expressed on myeloid cells 2 (Trem2) genes was significantly suppressed by HJ treatment. In addition, HJ administration significantly improved novel object recognition in a scopolamine-induced amnesia mouse model. CONCLUSIONS: These findings revealed that HJ has a beneficial effect on cognitive impairment and neuroinflammation induced by systemic inflammation and on amnesia induced by scopolamine in mice.
RESUMEN
The dopamine precursor 3,4dihydroxyphenyl lalanine (LDOPA) is the most widely used symptomatic treatment for Parkinson's disease (PD); however, its prolonged use is associated with LDOPAinduced dyskinesia in more than half of patients after 10 years of treatment. The present study investigated whether cotreatment with ßLapachone, a natural compound, and LDOPA has protective effects in a 6hydroxydopamine (6OHDA)induced mouse model of PD. Unilateral 6OHDAlesioned mice were treated with vehicle or ßLapachone (10 mg/kg/day) and LDOPA for 11 days. Abnormal involuntary movements (AIMs) were scored on days 5 and 10. ßLapachone (10 mg/kg) cotreatment with LDOPA decreased the AIMs score on both days 5 and 10. ßLapachone was demonstrated to have a beneficial effect on the axial and limb AIMs scores on day 10. There was no significant suppression in dopamine D1 receptorrelated and ERK1/2 signaling in the DAdenervated striatum by ßLapachonecotreatment with LDOPA. Notably, ßLapachonecotreatment with LDOPA increased phosphorylation at the Ser9 site of glycogen synthase kinase 3ß (GSK3ß), indicating suppression of GSK3ß activity in both the unlesioned and 6OHDAlesioned striata. In addition, astrocyte activation was markedly suppressed by ßLapachonecotreatment with LDOPA in the striatum and substantia nigra of the unilateral 6OHDA model. These findings suggest that ßLapachone cotreatment with LDOPA therapy may have therapeutic potential for the suppression or management of the development of LDOPAinduced dyskinesia in patients with PD.
Asunto(s)
Discinesias , Levodopa/efectos adversos , Naftoquinonas/farmacología , Oxidopamina/efectos adversos , Enfermedad de Parkinson Secundaria , Animales , Discinesias/tratamiento farmacológico , Discinesias/metabolismo , Discinesias/patología , Levodopa/farmacología , Masculino , Ratones , Oxidopamina/farmacología , Enfermedad de Parkinson Secundaria/inducido químicamente , Enfermedad de Parkinson Secundaria/tratamiento farmacológico , Enfermedad de Parkinson Secundaria/metabolismo , Enfermedad de Parkinson Secundaria/patologíaRESUMEN
Alzheimer's disease (AD) is the most prevalent neurodegenerative disease, and is characterized by the accumulation and presence of amyloid plaques (Aß), tangles, dementia, and cognitive impairment. Currently, there is no known cure for AD; however, recently, the association between alteration of the gut microbiota and AD pathology has been explored to find novel therapeutic approaches. Microbiota-targeted intervention has been suggested as an attractive therapeutic approach for AD. Agathobaculum butyriciproducens (SR79) is a strict anaerobic and butyric acid-producing bacteria. We hypothesized that administration of SR79 might have a beneficial effect on cognitive deficits and AD pathologies. To determine the therapeutic effects of SR79 on AD pathologies, APP/PS1 transgenic and lipopolysaccharide -induced cognitive impairment mouse models were used. In the lipopolysaccharide -induced cognitive deficit model, the administration of SR79 improved cognitive function and decreased microglia activation. In addition, the administration of SR79 to APP/PS1 mice significantly improved novel object recognition and percent alteration results in novel object recognition and Y-maze alteration tests. Furthermore, Aß plaque deposition and microglial activation were markedly reduced in the parietal cortex and hippocampus after SR79 treatment in APP/PS1 mice. SR79 treatment significantly decreased gene expression levels of IL-1ß and C1QB and increased the gene expression levels of IGF-1 and thereby the downstream signaling pathway in the cortex of APP/PS1 mice. In conclusion, SR79 administration improved cognitive function and AD pathologies through the regulation of neuroinflammation and IGF-1 signaling in an animal model.
Asunto(s)
Enfermedad de Alzheimer/terapia , Clostridiales/fisiología , Cognición , Disfunción Cognitiva/terapia , Microbioma Gastrointestinal/fisiología , Probióticos , Enfermedad de Alzheimer/microbiología , Péptidos beta-Amiloides/metabolismo , Animales , Encéfalo/patología , Encéfalo/fisiopatología , Modelos Animales de Enfermedad , Humanos , Factor I del Crecimiento Similar a la Insulina/genética , Factor I del Crecimiento Similar a la Insulina/metabolismo , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Lipopolisacáridos , Aprendizaje por Laberinto , Ratones , Ratones Transgénicos , Microglía/fisiología , Proteínas Mitocondriales/genética , Proteínas Mitocondriales/metabolismo , Placa Amiloide/terapia , Reconocimiento en PsicologíaRESUMEN
Humulus japonicus (HJ) is a widely used herbal medicine in Asia with antioxidative, antimicrobial, and antiinflammatory effects. We investigated the potential therapeutic effects of HJ in rheumatoid arthritis (RA) using a mouse model of collageninduced arthritis (CIA) and a lipopolysaccharidestimulated murine macrophage cell line (RAW 264.7). The CIA mice were administered 300 mg/kg HJ orally starting 3 days prior to second immunization. The clinical and histopathological findings were assessed in the paw of CIA mice. The levels of autoantibodies and inflammatory markers were determined in the plasma and cell culture supernatant, respectively. The expression at mRNA and protein levels was analyzed by reverse transcription quantitativePCR and western blot analysis, respectively. HJ significantly decreased the gross arthritic scores and paw swelling in CIA mice. Furthermore, synovial inflammation, cartilage destruction, and bone erosion were markedly reduced by HJ. It also decreased the expression of inflammatory enzymes in both the paw of mice and RAW 264.7 cells. Moreover, the expression of genes related to all macrophages and proinflammatory M1 macrophage were significantly decreased, whereas the expression of antiinflammatory M2 macrophage marker was markedly increased in the paw of HJtreated CIA mice. In addition, HJ suppressed the levels of plasma antitype II collagen antibody following the decreased expression of T helper type 1 (Th1) and Th2 cellassociated surface markers and cytokines in the paw. HJ also significantly inhibited the expression of IL6 both in vitro and in vivo, followed by reduced STAT3 phosphorylation and expression in the paw of CIA mice. Finally, the expression of osteoclastrelated genes was decreased in the paw of HJtreated CIA mice. These findings suggest that HJ can play a role in suppressing the development of CIA by overall regulation of articular inflammation. This study should provide new insights into the use of HJ as a therapeutically effective natural product against RA.
Asunto(s)
Antiinflamatorios/uso terapéutico , Artritis Experimental/tratamiento farmacológico , Artritis Reumatoide/tratamiento farmacológico , Humulus , Extractos Vegetales/uso terapéutico , Animales , Antiinflamatorios/química , Artritis Experimental/inmunología , Artritis Reumatoide/inmunología , Autoanticuerpos/inmunología , Modelos Animales de Enfermedad , Humulus/química , Inflamación/tratamiento farmacológico , Inflamación/inmunología , Mediadores de Inflamación/inmunología , Masculino , Ratones , Extractos Vegetales/química , Células RAW 264.7RESUMEN
Livestock infectious diseases, such as foot-and-mouth disease (FMD), cause substantial economic damage to livestock farms and their related industries. Among various causes of disease spread, airborne dispersion has previously been considered to be an important factor that could not be controlled by preventive measures to stop the spread of disease that focus on direct and indirect contact. Forecasting and predicting airborne virus spread are important to make time for developing strategies and to minimise the damage of the disease. To predict the airborne spread of the disease a modelling approach is important since field experiments using sensors are ineffective because of the rarefied concentrations of virus in the air. The simulation of airborne spread during past outbreaks required improvement both for farmers and for policy decision makers. In this study a free license computational fluid dynamics (CFD) code was used to simulate airborne virus spread. Forecasting data from the Korea Meteorological Administration (KMA) was directly connected in the developed model for real-time forecasting for 48 h in three-hourly intervals. To reduce computation time, scalar transport for airborne virus spread was simulated based on a database for the CFD computed airflow in the investigated area using representative wind conditions. The simulation results, and the weather data were then used to make a database for a web-based forecasting system that could be accessible to users.