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BACKGROUND: Porcine reproductive and respiratory syndrome virus (PRRSV) infection causes severe inflammatory response, respiratory disease and sow reproductive failure. Quercetin is among the widely occurring polypheno found abundantly in nature. Quercetin has anti-inflammatory, anti-oxidative and anti-viral properties. OBJECTIVES: This study aimed to explore the effect and mechanism of quercetin on PRRSV-induced inflammation in MARC-145 cells. METHODS: Observing the cytopathic effect and measurements of inflammatory markers in MARC-145 cells collectively demonstrate that quercetin elicits a curative effect on PRRSV-induced inflammation. Liquid chromatography-mass spectrometry was further used for a non-targeted metabolic analysis of the role of quercetin in the metabolic regulation of PRRSV inflammation in MARC-145 cells. RESULTS: It was shown that quercetin attenuated PRRSV-induced cytopathy in MARC-145 cells. Quercetin treatment inhibited PRRSV replication in MARC-145 cells in a dose-dependent manner. We also found that quercetin inhibited PRRSV-induced mRNA expression and secretion levels of tumour necrosis factor-α, interleukin 1ß and interleukin 6. Metabolomics analysis revealed that quercetin ameliorated PRRSV-induced inflammation. Pathway analysis results revealed that PRRSV-induced pathways including arachidonic acid metabolism, linoleic acid, glycerophospholipid and alanine, aspartate and glutamate metabolism were suppressed by quercetin. Moreover, we confirmed that quercetin inhibited the activation of NF-κB/p65 pathway, probably by attenuating PLA2, ALOX and COX mRNA expression. CONCLUSIONS: These results provide a crucial insight into the molecular mechanism of quercetin in alleviating PRRSV-induced inflammation.
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Ácido Araquidónico , Glutamina , Inflamación , Virus del Síndrome Respiratorio y Reproductivo Porcino , Quercetina , Quercetina/farmacología , Virus del Síndrome Respiratorio y Reproductivo Porcino/fisiología , Virus del Síndrome Respiratorio y Reproductivo Porcino/efectos de los fármacos , Animales , Línea Celular , Inflamación/virología , Inflamación/tratamiento farmacológico , Glutamina/metabolismo , Glutamina/farmacología , Ácido Araquidónico/metabolismo , Porcinos , Síndrome Respiratorio y de la Reproducción Porcina/virología , Síndrome Respiratorio y de la Reproducción Porcina/tratamiento farmacológico , Chlorocebus aethiopsRESUMEN
Terpinen-4-ol (T-4-O) is an important component of tea tree oil and has anti-inflammatory effects. Currently, there are very few studies on the mechanisms by which T-4-O improves lipopolysaccharide (LPS)-induced macrophage inflammation. In this study, LPS-stimulated mouse RAW264.7 macrophages were used as a model to analyze the effects of T-4-O on macrophage inflammatory factors and related metabolic pathways in an inflammatory environment. The results showed that T-4-O significantly decreased the expression levels of inflammatory cytokines induced by LPS. Cellular metabolism results showed that T-4-O significantly decreased the ratio of the extracellular acidification rate and oxygen consumption rate. Non-targeted metabolomics results showed that T-4-O mainly affected glutamine and glutamate metabolism and glycine, serine, and threonine metabolic pathways. qPCR results showed that T-4-O increased the transcript levels of GLS and GDH and promoted glutamine catabolism. Western blotting results showed that T-4-O inhibited the mTOR and IκB, thereby decreasing NF-κB activity. The overall results showed that T-4-O inhibited mTOR phosphorylation to promote glutamine metabolism and increased cell oxidative phosphorylation levels, thereby inhibiting the expression of LPS-induced inflammatory cytokines.
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Introduction: Trifolium pratense L. has anti-inflammatory, antioxidant, cardiovascular disease prevention, and estrogen-like effects. The existing method for the assay of effective components is commonly based on a spectrophotometer, which could not meet the requirement of quality control. Furthermore, although there have been many studies on the anti-inflammation effect of red clover, a few have been reported on the regulatory effect of red clover isoflavones (RCI) on lipopolysaccharide (LPS)-induced inflammatory response in porcine alveolar macrophages (3D4/2 cells), and its mechanism of action is still unclear. Methods: The main components of RCI including daidzein, genistein, and biochanin A were accurately quantified by high-performance liquid chromatography coupled with diode array detection (HPLC-DAD) after optimizing the extraction process through response surface methodology. The anti-inflammatory potential of RCI was carried out by detecting the level of inflammatory cytokines and mRNA expression of related genes. Furthermore, its anti-inflammatory mechanism was explored by investigating two signaling pathways (NF-κB and MAPK). Results: The optimal extraction conditions of RCI were as follows: the concentration of ethanol is 86% and the solid-liquid ratio is 1:29, with the herb particle size of 40 mesh sieve. Under the optimal conditions, the total extraction of target components of RCI was 2,641.469 µg/g. The RCI could significantly suppress the production and expression of many pro-inflammatory cytokines. The results of the Western blot revealed that RCI dramatically reduced the expression of p65, p-p65, IκB-α, p38, and p-p38. These results are associated with the suppression of the signal pathway of p38 MAPK, and on the contrary, activating the NF-κB pathway. Collectively, our data demonstrated that RCI reversed the transcription of inflammatory factors and inhibited the expression of p65, p-p65, IκB-α, and p38, indicating that RCI had excellent anti-inflammatory properties through disturbing the activation of p38 MAPK and NF-κB pathways. Conclusion: The extraction conditions of RCI were optimized by HPLC-DAD combined with response surface methodology, which will contribute to the quality control of RCI. RCI had anti-inflammatory effects on the LPS-induced 3D4/2 cells. Its mechanism is to control the activation of NF-κB and p38 MAPK pathways, thereby reducing the expression of inflammatory-related genes and suppressing the release of cytokines.
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Alcoholic liver disease represents a serious threat to human health. In terms of safety and acceptability, thymol is widely used in or on foodstuffs to generate odour and taste. The present study aimed to investigate the therapeutic effect and mechanism of thymol against ethanol-induced injury in liver cells. Here we found that thymol is an effective agent for reducing ethanol-induced reactive oxygen species production in mouse liver cells. Thymol improves ethanol-induced lipid accumulation, and this corresponded to altered DGAT2 mRNA expression levels. Metabolomics data analysis showed that thymol alleviated ethanol-induced changes in the levels of thirty-four metabolites including nicotinic acid and l-arginine. By utilizing pathway enrichment analysis, altered metabolites in cells treated with ethanol and ethanol plus thymol were enriched in fourteen pathways including metabolic pathways and arginine and proline metabolism. We further confirmed the alleviation of overdose nitric oxide production in cells treated with ethanol plus thymol compared with that in ethanol-treated cells. It was interesting that up-regulated LC3-II/LC3-I ratio together with higher SQSTM1 protein abundance in ethanol-treated cells were attenuated by treatment with ethanol plus thymol. Thymol ameliorated ethanol-induced reduction of HSPA8 protein abundance. In addition, chloroquine-treated cells exhibited lower HSPA8 protein abundance compared with cells simulated with ethanol plus thymol. These data reveal that improving effect of thymol on ethanol-induced metabolic alteration is related to autophagic flux restoration. Our findings indicate that thymol is an attractive option for treating ethanol-induced liver damage.
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Enfermedad Hepática Inducida por Sustancias y Drogas , Hepatopatías Alcohólicas , Ratones , Animales , Humanos , Etanol/toxicidad , Timol/farmacología , Timol/uso terapéutico , Autofagia , Enfermedad Hepática Inducida por Sustancias y Drogas/tratamiento farmacológico , Enfermedad Hepática Inducida por Sustancias y Drogas/prevención & control , HígadoRESUMEN
In order to investigate the apoptosis triggered by porcine circovirus type 2 (PCV2) in lymphocytes and the underlying mechanism, the levels of apoptosis and the expression levels of miRNA were examined by flow cytometry, Western blotting and real-time PCR (qPCR). The mimics or inhibitors of miR-125a-5p, an apoptosis-related miRNA, were transfected into PK-15 cells, and the apoptosis rate was examined upon overexpression or inhibition of mir-125a-5p. The target gene of mir-125a-5p was predicted by bioinformatics method, and the regulation of mir-125a-5p on the target gene was analyzed by luciferase reporter assay. The expressions of Bcl-2, Bax, cytochrome C and caspase-3 were detected by Western blotting. The results showed that exosomes secreted by PK-15 cells infected with PCV2 significantly increased the lymphocyte apoptosis rate, which was dose-dependent in certain concentration range. The expression of miR-125a-5p was dramatically increased. The apoptosis rate was increased significantly in the cells transfected with miR-125a-5p. It was predicted that there were binding sites of miR-125a-5p at Bcl-2 3'UTR by TargetScan. The luciferase activity of wild-type pmir-Bcl-2 3'UTR was inhibited significantly by miR-125a-5p mimics, but that of mutant pmir-Bcl-2 3'UTR was not changed. By Western blotting, Bcl-2 was reduced significantly, while Bax, cytochrome C and caspase-3 increased significantly, and the ratio of Bcl-2/Bax was significantly decreased. These results showed that PCV2 up-regulated the expression of miR-125a-5p through exosomes, then inhibited the expression of Bcl-2 at both mRNA and protein level, activated mitochondrial apoptosis pathway and induced apoptosis in lymphocytes.
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Circovirus , MicroARNs , Regiones no Traducidas 3' , Animales , Apoptosis/genética , Caspasa 3/genética , Proliferación Celular/genética , Circovirus/genética , Citocromos c/genética , Luciferasas/genética , Linfocitos/metabolismo , MicroARNs/genética , MicroARNs/metabolismo , Porcinos , Proteína X Asociada a bcl-2/genéticaRESUMEN
Klebsiella infection is widely acknowledged to inflict severe inflammatory damage in bovines. Herein, we demonstrate significant death of EpH4-Ev cells incubated with Klebsiella. And compelling evidence shows that Klebsiella infection increases interactions between the Receptor-interacting serine/threonine-protein kinase 1 (RIPK1) and RIPK3, which promotes phosphorylation of RIPK3 and MLKL to induce necroptosis. However, these changes can be partially reversed by taurine and Nec-1s. Moreover, using taurine and Nec-1s to partially inhibit necroptosis significantly reduce TNF-α, IL-1ß and IL-6 levels and NAGase activity induced by Klebsiella infection. Taken together, taurine partially inhibits necroptosis induced by Klebsiella infection and hence alleviates inflammatory and injury in EpH4-Ev cells.
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Enfermedades de los Bovinos , Infecciones por Klebsiella , Animales , Apoptosis , Bovinos , Infecciones por Klebsiella/tratamiento farmacológico , Infecciones por Klebsiella/veterinaria , Necroptosis , Fosforilación , Proteínas Quinasas/metabolismo , Proteína Serina-Treonina Quinasas de Interacción con Receptores/metabolismo , TaurinaRESUMEN
The study evaluated prebiotic potential and the enzyme inhibition of extracts and isolated compounds of Rumex vesicarius (ruby dock), family Polygonaceae. Eight known compounds were identified in the roots of R. vesicarius. Extracts and compounds (1-8) increased the growth rate of Escherichia coli Nissle 1917 differentially compared to controls. The highest prebiotic index (PI) and activity score was recorded for EcN in the presence of compound 4, followed by, in descending order, petroleum ether, ethyl acetate, and total methanol extracts. The compounds and extracts reduced protease, α-amylase, and angiotensin-converting enzyme activities. This inhibitory activity was positively correlated with PI, Pscore, µu, and Ymax. These findings suggest that R. vesicarius is a good source of potential prebiotic and can boost beneficial bacteria. It may also be considered promising for treatment of diabetes mellitus, controlling weight, and regulating blood pressure.
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For the purpose of efficient targeted therapies, suppressing phagocytosis by a mononuclear phagocyte system (MPS), enhancing the "active" targeted delivery, and meeting clinical production criteria are extremely critical for engineering strategies of novel drug delivery systems. Herein, we used a chemically-induced membrane blebbing and extrusion combined method to induce triple-negative breast cancer (TNBC) cell apoptosis to secrete apoptotic body analogue (ABA) vesicles on a large scale for therapeutic drug delivery. After optimization, the ABAs have a desirable size, good biocompatibility, and long-term colloidal stability. Furthermore, ABAs present anti-phagocytosis ("don't eat me") and specific homologous targeting ("eat me") capacities because of their inheritance of membrane proteins such as CD47 and cellular adhesion molecules from parent cells. After loading with toxic protein saporin and anti-twist siRNA, ABAs can significantly inhibit the growth and lung metastasis of TNBC in an orthotopic metastasis model due to their reduced clearance of immune organs, long circulation time, and enhanced targeted accumulation at the tumor sites. These results suggest the great potential of ABAs for targeted drug delivery therapy, in particular efficient TNBC treatment.
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Apoptosis , Sistemas de Liberación de Medicamentos , Sistema Mononuclear Fagocítico/fisiología , Neoplasias de la Mama Triple Negativas/terapia , Animales , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Vesículas Extracelulares , Humanos , Ratones , Ratones Endogámicos ICR , Ratones Desnudos , Nanoestructuras , Fagocitosis , Células RAW 264.7 , Distribución Aleatoria , Saporinas/química , Saporinas/farmacología , Distribución Tisular , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
The chromatographic reinvestigation the methanol extract of Tetraena aegyptia led to the separation of a new flavonoid glycoside, isorhamnetin-3-O-[2```,3```-O-isopropylidene-α-L-rhamnopyranosyl]-(1```â6``)-O-ß-D-glucopyranoside (1), together with two known flavonoids, isorhamnetin (2) and isorhamnetin-3-O-glucoside (3), isolated for the first time from the plant. The new compound was evaluated for the anti-inflammatory activity by using LPS-induce RAW 264.7 cells model. Compound 1 showed significant inhibitory effect on NO release. ELISA assay showed a pronounced effect of 1 on the secretion of cytokines IL-6 and TNF-α, in a dose-dependent manner. Consistent results were obtained by qRT-PCR which revealed that compound 1 markedly reduced the mRNA expression of IL-6 and TNF-α. Together these data, we demonstrated the anti-inflammatory activity of compound 1.
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Antiinflamatorios no Esteroideos/química , Antiinflamatorios no Esteroideos/farmacología , Flavonoides/química , Flavonoides/farmacología , Zygophyllaceae/química , Animales , Antiinflamatorios no Esteroideos/administración & dosificación , Relación Dosis-Respuesta a Droga , Evaluación Preclínica de Medicamentos , Flavonoles/aislamiento & purificación , Flavonoles/farmacología , Interleucina-6/genética , Interleucina-6/metabolismo , Lipopolisacáridos/toxicidad , Ratones , Estructura Molecular , Óxido Nítrico/metabolismo , Quercetina/análogos & derivados , Quercetina/aislamiento & purificación , Quercetina/farmacología , Células RAW 264.7 , Factor de Necrosis Tumoral alfaRESUMEN
Continued chromatographic investigation of Calendula officinalis flowers led to the isolation of two sesquiterpenes, including one new, viridiflorol-10-O-ß-quinovopyranoside-2`-O-(3``-methyl-2``-pentenoate) (1), along with a previously reported compound viridiflorol-10-O-ß-fucopyranoside-2`-O-(3``-methyl-2``-pentenoate) (2). The new compound 1 was tested for antiprotozoal activity against Leishmania donovani Amastigote/THP1 and Trypanosoma brucei and it showed IC50 values of 3.57 and 7.84 µg/mL, respectively, while compound 2 exhibited no activity at the highest concentration tested 10 µg/mL.
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Antiprotozoarios , Calendula , Leishmania donovani , Sesquiterpenos , Antiprotozoarios/farmacología , Ésteres , Flores , Glicósidos/farmacología , Extractos Vegetales/farmacología , Sesquiterpenos/farmacologíaRESUMEN
Recent studies have demonstrated that rosmarinic acid is a valuable natural product for treatment of alcoholic liver disease. However, the mechanisms whereby rosmarinic acid improves alcoholic liver disease remain unclear. Here we performed experiments using a non-transformed mouse hepatocyte cell line (AML12). Oil-red O staining demonstrated that rosmarinic acid reduced ethanol-induced lipid accumulation. It was shown that rosmarinic acid prevented ethanol-induced elevation of the malondialdehyde level. We also found that rosmarinic acid inhibited ethanol-induced mRNA expression of tumor necrosis factor-α and interleukin 6. Metabolomics analysis revealed that rosmarinic acid ameliorated ethanol-induced fatty acid biosynthesis in the cytoplasm. In addition, palmitic acid was a candidate biomarker in cells exposed to ethanol or ethanol plus rosmarinic acid. Rosmarinic acid prevented the ethanol-induced increase in sorbitol that is a component of the polyol pathway. Moreover, we confirmed that rosmarinic acid attenuated ethanol-induced mRNA expression of fatty acid synthase, probably by modulating the AMPK/SREBP-1c pathway. Furthermore, rosmarinic acid prevented the ethanol-induced decrease in eight metabolites that are involved in mitochondrial metabolism, including glycine and succinic acid which are the components of carnitine synthesis. These results provide a crucial insight into the molecular mechanism of rosmarinic acid in alleviating ethanol-induced injury.
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Cinamatos/farmacología , Depsidos/farmacología , Etanol/farmacología , Ácidos Grasos/biosíntesis , Hepatocitos/efectos de los fármacos , Animales , Línea Celular , Hepatocitos/metabolismo , Metabolismo de los Lípidos , Ratones , Estrés Fisiológico , Ácido RosmarínicoRESUMEN
In the present study, the safety of red clover isoflavone extract (RCIE) and its potential adjuvant effects on the cellular and humoral immune responses to ovalbumin (OVA) were evaluated using an ICR mouse model. On day 1, the mice were first subcutaneously immunized with 100 µg OVA, 100 µg OVA + 200 µg aluminum hydroxide gel (alum) or OVA + 50, 100 or 200 µg RCIE (RCIE + OVA), following which booster immunization was performed on day 15. After 2 weeks, the stimulation of splenocyte proliferation and levels of serum antibodies were measured. No notable stress responses were observed after the initial and booster immunization. Splenocyte proliferation was significantly increased in mice immunized with OVA + 100 µg RCIE (P<0.01). The levels of IgG, IgG1 and IgG2a antibodies in serum were also significantly increased in OVA + RCIE groups compared with the OVA control group (P<0.05). In the OVA + RCIE groups, serum levels of interleukin (IL)-2, interferon-γ (IFN-γ) and IL-10 were increased, and the mRNA expression levels of IL-2, IFN-γ, IL-4, IL-10, T-bet and GATA-3 were also significantly increased compared with the OVA control group (P<0.05) in splenocytes. In addition, as an adjuvant, RCIE significantly increased the survival rates of mice inoculated with an E. coli vaccine and enhanced the early immune protection against pathogenic E. coli. In conclusion, these findings suggest that RCIE can be used as a safe vaccine adjuvant and supports its use in clinical applications.
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Machaerinic acid 3-O-ß-D-glucuronopyranoside (1), along with ten known compounds (2-11) were isolated from the methanol extract of Calendula officinalis L. aerial parts. Their structures were confirmed by 1D and 2D NMR analysis and HRESIMS. Compound 1 was evaluated for the anti-proliferative activity against 95D and HT29 cancer cell lines and showed no cytotoxicity at the concentration of 100 µM.
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Antineoplásicos Fitogénicos/química , Antineoplásicos Fitogénicos/farmacología , Calendula/química , Antineoplásicos Fitogénicos/aislamiento & purificación , Células HT29 , Humanos , Espectroscopía de Resonancia Magnética , Estructura Molecular , Componentes Aéreos de las Plantas/química , Extractos Vegetales/química , Espectrometría de Masa por Ionización de ElectrosprayRESUMEN
Aldose reductase (AR) has been reported to be involved in the development of nonalcoholic fatty liver disease (NAFLD). Hepatic AR is induced under hyperglycemia condition and converts excess glucose to lipogenic fructose, which contributes in part to the accumulation of fat in the liver cells of diabetes rodents. In addition, the hyperglycemia-induced AR or nutrition-induced AR causes suppression of the transcriptional activity of peroxisome proliferator-activated receptor (PPAR) α and reduced lipolysis in the liver, which also contribute to the development of NAFLD. Moreover, AR induction in non-alcoholic steatohepatitis (NASH) may aggravate oxidative stress and the expression of inflammatory cytokines in the liver. Here, we summarize the knowledge on AR inhibitors of plant origin and review the effect of some plant-derived AR inhibitors on NAFLD/NASH in rodents. Natural AR inhibitors may improve NAFLD at least in part through attenuating oxidative stress and inflammatory cytokine expression. Some of the natural AR inhibitors have been reported to attenuate hepatic steatosis through the regulation of PPARα-mediated fatty acid oxidation. In this review, we propose that the natural AR inhibitors are potential therapeutic agents for NAFLD.
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Aldehído Reductasa/antagonistas & inhibidores , Aldehído Reductasa/metabolismo , Inhibidores Enzimáticos/farmacología , Inhibidores Enzimáticos/uso terapéutico , Enfermedad del Hígado Graso no Alcohólico/tratamiento farmacológico , Fitoterapia , Extractos Vegetales/química , Extractos Vegetales/farmacología , Animales , Humanos , Metabolismo de los Lípidos/efectos de los fármacos , Enfermedad del Hígado Graso no Alcohólico/prevención & control , Extractos Vegetales/uso terapéuticoRESUMEN
Alcoholic liver disease (ALD) is caused by heavy alcohol consumption over a long period. Acetaldehyde-mediated toxicity, oxidative stress, and imbalance of lipid metabolism are generally considered involved in the initiation of ALD. There is an increasing requirement for alternative and natural medicine to treat ALD. Recently, aldose reductase (AR) has been reported to be involved in the development of ALD by affecting inflammatory cytokines, oxidative stress, and lipid metabolism. Here, we review the effect of plant-derived AR inhibitors on ALD in rodents. And we conclude that AR inhibitors of plant origin may enhance antioxidant capacity, inhibit lipid peroxidation and inflammatory cytokines expression, and activate AMP-activated protein kinase thereby subsequently suppressing alcohol-induced lipid synthesis in liver to achieve ALD protection. This review reveals that natural AR inhibitor may be potential therapeutic agent for ALD.
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Aldehído Reductasa/antagonistas & inhibidores , Inhibidores Enzimáticos/farmacología , Hepatopatías Alcohólicas/tratamiento farmacológico , Plantas/química , Aldehído Reductasa/química , Antioxidantes/química , Antioxidantes/farmacología , Inhibidores Enzimáticos/química , Humanos , Metabolismo de los Lípidos/efectos de los fármacos , Hígado/química , Hígado/efectos de los fármacos , Estrés Oxidativo , Transducción de Señal/efectos de los fármacosRESUMEN
SCOPE: Alcoholic liver disease is a serious threat to human health. The development of drug candidates from complementary and alternative medicines is an attractive approach. Myricetin can be found in fruit, vegetables, and herbs. This study investigates the protective effect of myricetin on ethanol-induced injury in mouse liver cells. METHODS AND RESULTS: Oil-red O staining, assays of oxidative stress and measurements of inflammatory markers in mouse AML12 liver cells collectively demonstrate that myricetin elicits a curative effect on ethanol-induced injury. Next, the role of myricetin in the metabolic regulation of ethanol pathology in liver cells is assessed by gas chromatography coupled with mass spectrometry. Myricetin inhibits ethanol-stimulated fatty acid biosynthesis. Additionally, dodecanoic acid may be proposed as a potential biomarker related to ethanol pathology or myricetin therapy. It is also observed that myricetin enhances ethanol-induced inhibition of the mitochondrial electron transport chain. Moreover, fumaric acid is found to be a candidate biomarker related to ethanol toxicity or myricetin therapy. Quantitative reverse-transcription-PCR shows that ethanol-induced fatty acid synthase and sterol regulatory element-binding protein-1c mRNA levels are alleviated by myricetin. Finally, myricetin increases ethanol-induced inhibition of phosphorylation of AMP-activated protein kinase. CONCLUSION: These results elucidate the pharmacological mechanism of myricetin on ethanol-induced lipid accumulation.
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Etanol/toxicidad , Ácidos Grasos/biosíntesis , Flavonoides/farmacología , Hepatocitos/efectos de los fármacos , Hepatocitos/metabolismo , Metabolismo de los Lípidos/efectos de los fármacos , Animales , Línea Celular , Cromatografía de Gases y Espectrometría de Masas , Regulación de la Expresión Génica/efectos de los fármacos , Metaboloma/efectos de los fármacos , Ratones , Análisis Multivariante , Especies Reactivas de Oxígeno/metabolismo , Proteína 1 de Unión a los Elementos Reguladores de Esteroles/genética , Proteína 1 de Unión a los Elementos Reguladores de Esteroles/metabolismoRESUMEN
Alcoholic liver injury results in morbidity and mortality worldwide, but there are currently no effective and safe therapeutics. Previously we demonstrated that aldose reductase (AR) inhibitor ameliorated alcoholic hepatic steatosis. To clarify the mechanism whereby AR inhibitor improves alcoholic hepatic steatosis, herein we investigated the effect of AR inhibitor on hepatic metabolism in mice fed a Lieber-DeCarli liquid diet with 5% ethanol. Nontargeted metabolomics showed carbohydrates and lipids were characteristic categories in ethanol diet-fed mice with or without AR inhibitor treatment, whereas AR inhibitor mainly affected carbohydrates and peptides. Ethanol-induced galactose metabolism and fatty acid biosynthesis are important for the induction of hepatic steatosis, while AR inhibitor impaired galactose metabolism without perturbing fatty acid biosynthesis. In parallel with successful treatment of steatosis, AR inhibitor suppressed ethanol-activated galactose metabolism and saturated fatty acid biosynthesis. Sorbitol in galactose metabolism and stearic acid in saturated fatty acid biosynthesis were potential biomarkers responsible for ethanol or ethanol plus AR inhibitor treatment. In vitro analysis confirmed that exogenous addition of sorbitol augmented ethanol-induced steatosis and stearic acid. These findings not only reveal metabolic patterns associated with disease and treatment, but also shed light on functional biomarkers contribute to AR inhibition therapy.
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Aldehído Reductasa/antagonistas & inhibidores , Inhibidores Enzimáticos/farmacología , Ácidos Grasos/biosíntesis , Hígado Graso Alcohólico/prevención & control , Metaboloma/efectos de los fármacos , Aldehído Reductasa/metabolismo , Animales , Biomarcadores/metabolismo , Línea Celular , Análisis Discriminante , Inhibidores Enzimáticos/uso terapéutico , Etanol/toxicidad , Hígado Graso Alcohólico/patología , Hígado Graso Alcohólico/veterinaria , Análisis de los Mínimos Cuadrados , Hígado/metabolismo , Hígado/patología , Masculino , RatonesRESUMEN
It is necessary to identify compounds that may provide protection against alcoholic liver disease. To the best of our knowledge, the effect of myricitrin on the development of ethanolinduced liver disease has not been previously investigated. The present study aimed to determine the effect of myricitrin on ethanolinduced steatosis in AML12 mouse liver cells and to identify the underlying molecular mechanisms. Ethanoltreated AML12 cells exhibited significant improvement in viability following treatment with myricitrin. Oil red O staining indicated that myricitrin ameliorated ethanolinduced lipid accumulation in cells. Furthermore, following treatment with myricitrin, improvement in ethanolinduced steatosis and decrease in the levels of reactive oxygen species and lipoperoxides were observed in ethanolstimulated cells. Myricitrin suppressed mRNA and protein expression of tumor necrosis factorα, interleukin6 and transforming growth factorß1 in ethanolstimulated AML12 cells. Myricitrin markedly increased phosphorylation of adenosine monophosphateactivated protein kinase (AMPK) and significantly reduced mRNA expression of sterolregulatory elementbinding protein1c (SREBP1c) and fatty acid synthase in ethanolstimulated AML12 cells. The results of the present study indicate that myricitrin ameliorates ethanolinduced steatosis in AML12 cells by attenuating oxidative stress, suppressing expression of certain inflammatory cytokines and modulating the AMPK/SREBP-1c pathway.
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Antioxidantes/uso terapéutico , Activadores de Enzimas/uso terapéutico , Hígado Graso Alcohólico/tratamiento farmacológico , Flavonoides/uso terapéutico , Hígado/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Proteínas Quinasas/metabolismo , Quinasas de la Proteína-Quinasa Activada por el AMP , Animales , Línea Celular , Citocinas/análisis , Activación Enzimática/efectos de los fármacos , Etanol , Hígado Graso Alcohólico/metabolismo , Hígado Graso Alcohólico/patología , Hígado/metabolismo , Hígado/patología , RatonesRESUMEN
Transcription factor EB (TFEB) is a master regulator of autophagy and lysosomal biogenesis. Here, silver nanoparticles (Ag NPs)-induced cytoprotective autophagy required TFEB is shown. Ag NPs-induced nucleus translocation of TFEB through a well-established mechanism involving dephosphorylation of TFEB at serine-142 and serine-211 but independent of both the mTORC1 and ERK1/2 pathways. TFEB nucleus translocation precedes autophagy induced by Ag NPs and leads to enhanced expression of autophagy-essential genes. Knocking down the expression of TFEB attenuates the autophagy induction is demonstrated, and in the meantime, enhanced cell killing in HeLa cells treats with Ag NPs, indicating that TFEB is the key mediator for Ag NPs-induced cytoprotective autophagy. The results pinpoint TFEB as a potential target for developing more effective Ag NPs-based cancer therapeutics.
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Autofagia/efectos de los fármacos , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/metabolismo , Núcleo Celular/metabolismo , Nanopartículas del Metal/química , Plata/química , Plata/farmacología , Células HeLa , Humanos , Transporte de Proteínas/efectos de los fármacos , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
Gynura formosana Kitam (family Compositae) has traditionally been used for the prevention of diabetes, cancer and inflammation in China. However, there are few reports of its anti-inflammatory effects. In the present study, after assessing the in vitro antioxidant activities of extracts from Gynura formosana Kitam leaves, the anti-inflammatory and antioxidant activities of ethyl acetate extract of Gynura formosana Kitam leaves (EAEG) were further investigated in rats using a cotton pellet-induced model of granuloma. EAEG significantly inhibited the formation of cotton pellet-induced granuloma in rats in a dose-dependent manner. Moreover, EAEG treatment significantly decreased the level of plasma C-reactive protein and suppressed the activities of plasma glutamate pyruvate transaminase and lactate dehydrogenase in model groups. Furthermore, EAEG increased the hepatic levels of anti-oxidative enzymes or antioxidants, including catalase, superoxide dismutase and reduced glutathione, and reduced the level of lipid peroxidation in the rat model of inflammation. In addition, EAEG decreased plasma levels of tumor necrosis factor-α and interleukin-1ß in the rat model of inflammation. Therefore, our results indicated that EAEG had potent anti-inflammatory effects, which was achieved at least in part through activating antioxidant enzyme activities and suppressing the production of proinflammatory mediators by macrophages.