RESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Chuanminshen violaceum M. L. Sheh & R. H. Shan (CV) is used as a medicine with roots, which have the effects of benefiting the lungs, harmonizing the stomach, resolving phlegm and detoxifying. Polysaccharide is one of its main active components and has various pharmacological activities, but the structural characterization and pharmacological activities of polysaccharide from the stems and leaves parts of CV are still unclear. AIM OF THE STUDY: The aim of this study was to investigate the optimal extraction conditions for ultrasound-assisted extraction of polysaccharide from CV stems and leaves, and to carry out preliminary structural analyses, anti-inflammatory and antioxidant effects of the obtained polysaccharide and to elucidate the underlying mechanisms. MATERIALS AND METHODS: The ultrasonic-assisted extraction of CV stems and leaves polysaccharides was carried out, and the response surface methodology (RSM) was used to optimize the extraction process to obtain CV polysaccharides (CVP) under the optimal conditions. Subsequently, we isolated and purified CVP to obtain the homogeneous polysaccharide CVP-AP-I, and evaluated the composition, molecular weight, and structural features of CVP-AP-I using a variety of technical methods. Finally, we tested the pharmacological activity of CVP-AP-â in an LPS-induced model of oxidative stress and inflammation in intestinal porcine epithelial cells (IPEC-J2) and explored its possible mechanism of action. RESULTS: The crude polysaccharide was obtained under optimal extraction conditions and subsequently isolated and purified to obtain CVP-AP-â (35.34 kDa), and the structural characterization indicated that CVP-AP-â was mainly composed of galactose, galactose, rhamnose and glucose, which was a typical pectic polysaccharide. In addition, CVP-AP-â attenuates LPS-induced inflammation and oxidative stress by inhibiting the expression of pro-inflammatory factor genes and proteins and up-regulating the expression of antioxidant enzyme-related genes and proteins in IPEC-J2, by a mechanism related to the activation of the Nrf2/Keap1 signaling pathway. CONCLUSION: The results of this study suggest that the polysaccharide isolated from CV stems and leaves was a pectic polysaccharide with similar pharmacological activities as CV roots, exhibiting strong anti-inflammatory and antioxidant activities, suggesting that CV stems and leaves could possess the same traditional efficacy as CV roots, which is expected to be used in the treatment of intestinal diseases.
Asunto(s)
Antiinflamatorios , Antioxidantes , Hojas de la Planta , Tallos de la Planta , Polisacáridos , Hojas de la Planta/química , Polisacáridos/farmacología , Polisacáridos/aislamiento & purificación , Polisacáridos/química , Animales , Tallos de la Planta/química , Antiinflamatorios/farmacología , Antiinflamatorios/aislamiento & purificación , Antiinflamatorios/química , Antioxidantes/farmacología , Antioxidantes/aislamiento & purificación , Ratones , Porcinos , Extractos Vegetales/farmacología , Extractos Vegetales/química , Intestinos/efectos de los fármacos , Células RAW 264.7RESUMEN
The roots of Angelica sinensis have been used in Traditional Chinese Medicine for thousands of years. However, tons of aerial parts of this herb (aboveground part) are commonly discarded during the process of root preparations. A polysaccharide (ASP-Ag-AP) in the aboveground parts of A. sinensis was isolated and preliminarily characterized as typical plant pectin. ASP-Ag-AP exhibited noticeable protective effects against dextran sodium sulfate (DSS)-induced colitis, including reduction of colonic inflammation, modulation of barrier function, and alteration of gut microbiota and serum metabolite profile. Anti-inflammatory effects of ASP-Ag-AP were observed by inhibiting TLR4/MyD88/NF-κB signaling pathway in vitro and in vivo. Additionally, the level of serum metabolite 5-methyl-dl-tryptophan (5-MT) was reduced by DSS and restored by ASP-Ag-AP, which also negatively correlated with Bacteroides, Alistipes, Staphylococcus and pro-inflammatory factors. The protection from inflammatory stress on intestinal porcine enterocytes cells (IPEC-J2) of 5-MT was observed through the inhibition of TLR4/MyD88/NF-κB pathway. Besides, 5-MT also exhibited robust anti-inflammatory effect in colitis mice with improving colitis symptoms, barrier function and gut microbiota, which was the same as presented by ASP-Ag-AP. Therefore, ASP-Ag-AP could be a promising agent for colitis prevention and 5-MT could be the signal metabolite of ASP-Ag-AP on defending against intestinal inflammatory stress.
Asunto(s)
Angelica sinensis , Colitis , Microbioma Gastrointestinal , Ratones , Animales , Porcinos , FN-kappa B/metabolismo , Factor 88 de Diferenciación Mieloide/metabolismo , Angelica sinensis/metabolismo , Receptor Toll-Like 4/metabolismo , Colitis/inducido químicamente , Colitis/tratamiento farmacológico , Colitis/metabolismo , Polisacáridos/uso terapéutico , Antiinflamatorios/farmacología , Sulfato de Dextran/efectos adversos , Modelos Animales de EnfermedadRESUMEN
BACKGROUND: Molecular epidemiological studies have sought associations between Fat mass and obesity associated (FTO) gene polymorphisms and gestational diabetes mellitus (GDM) risk, but findings are inconsistent. Hence, we performed a meta-analysis to clarify this problem. METHODS: Case-control studies reporting the relationship of three FTO polymorphisms (rs9939609, rs8050136, and rs1421085) and GDM published before June 2018 were searched in 6 electronic databases such as PubMed and Embase. Pooled odds ratios (ORs) with 95% confidence intervals (CIs) were used to assess the strength of associations. Trial sequential analysis (TSA) was performed to evaluate the type 1 and type 2 errors. RESULTS: A total of 5 studies involving 703 GDM cases and 2700 controls for rs9939609, 3 studies involving 1144 GDM cases and 909 controls for rs8050136, and 2 studies involving 207 GDM cases and 205 controls for rs1421085, were included in the meta-analysis. No association was observed between the three polymorphisms with the GDM risk under all genetic models. For example, the ORs and its 95% CIs under dominant genetic model were 0.88 (0.59, 1.33) for rs9939609, 1.11 (0.91, 1.35) for rs8050136, and 0.91 (0.58, 1.41) for rs1421085, respectively. Under TSA, there are insufficient levels of evidence for all of these three polymorphisms. CONCLUSION: The present meta-analysis provides statistical evidence indicating a lack of association between FTO polymorphismsand GDM risk. More studies with larger sample size are needed to confirm these null associations.
Asunto(s)
Dioxigenasa FTO Dependiente de Alfa-Cetoglutarato/genética , Diabetes Gestacional/genética , Predisposición Genética a la Enfermedad/genética , Polimorfismo de Nucleótido Simple/genética , Animales , Estudios de Casos y Controles , Diabetes Mellitus Tipo 2/genética , Femenino , Humanos , Obesidad , Oportunidad Relativa , Embarazo , Factores de RiesgoRESUMEN
We previously reported the in vitro and in vivo hepatoprotective and anti-fibrotic effects of PF2401-SF, a standardized fraction of Salvia miltiorrhiza, against acute and subacute liver injury. The aim of this study was to investigate the effect of PF2401-SF on liver fibrosis induced by thioacetamide (TAA), a chronic liver injury model (12 weeks) that closely resembles fibrosis and cirrhosis in humans. Hepatoprotective activity was indicated by low serum levels of the markers aspartate amino transferase and alanine amino transferase .In addition, compared to the TAA-group livers, the PF2401-SF-treated liver tissues showed no fibrous tissue deposition in the portal areas, hepatocyte morphology more closely resembling normal tissue morphology, and significantly reduced collagen deposition. Furthermore, downregulation of collagen 1(α) and tissue inhibitor of metalloproteinase (TIMP)1 protein and mRNA expression also supports PF2401-SF's anti-fibrotic effect. We also observed reduced expression of α-smooth muscle actin (α-SMA), an important marker of hepatic stellate cells (HSCs) activation. From these results, we conclude that PF2401-SF's anti-fibrotic mechanism in the TAA model involves reduced HSC activation, and may be mediated by downregulation of central markers of fibrosis, including collagen 1(α), TIMP1, and α-SMA.
Asunto(s)
Medicamentos Herbarios Chinos/uso terapéutico , Cirrosis Hepática/inducido químicamente , Cirrosis Hepática/prevención & control , Extractos Vegetales/uso terapéutico , Salvia miltiorrhiza , Tioacetamida/toxicidad , Animales , Medicamentos Herbarios Chinos/aislamiento & purificación , Cirrosis Hepática/metabolismo , Masculino , Extractos Vegetales/aislamiento & purificación , Ratas , Ratas Sprague-Dawley , Resultado del TratamientoRESUMEN
Hepatic stellate cells (HSCs) play a central role in liver fibrosis. Inhibition of HSC growth and induction of apoptosis have been proposed as therapeutic strategies for the treatment and prevention of liver fibrosis. Propyl gallate (PG) is an antioxidant widely used in processed foods, cosmetics and medicinal preparations. However, the anti-fibrotic effect of PG in liver injury is unclear. In this study, we investigated whether PG could induce apoptosis in activated HSCs. Treatment of activated HSCs with PG inhibited cell viability in a dose- and time-dependent manner. PG induced apoptosis as demonstrated by morphological changes, poly(ADP-ribose) polymerase (PARP) cleavage, caspase-3 cleavage, increased Bad expression, and decreased Bcl-2 protein expression. Through stimulation of the activation of c-Jun NH2-terminal protein kinase (JNK) and p38 mitogen-activated protein kinases (MAPK) by PG treatment, we demonstrated that JNK and p38 MPAK are not involved in PG-induced apoptosis using their specific inhibitors. Taken together, these findings indicate that PG induces apoptosis in activated HSCs. The potential anti-fibrotic effect of PG warrants further evaluation.
Asunto(s)
Apoptosis/efectos de los fármacos , Células Estrelladas Hepáticas/efectos de los fármacos , Galato de Propilo/farmacología , Animales , Apoptosis/fisiología , Línea Celular Transformada , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/fisiología , Relación Dosis-Respuesta a Droga , Células Estrelladas Hepáticas/fisiología , Galato de Propilo/química , RatasRESUMEN
Suppression of hepatic stellate cell (HSC) activation and proliferation, and induction of apoptosis in activated HSCs have been proposed as therapeutic strategies for the treatment and prevention of the hepatic fibrosis. We previously showed that 2',4',6'-tris(methoxymethoxy) chalcone (TMMC), a synthesized chalcone derivative, inhibits platelet-derived growth factor-induced HSC proliferation at 5-20 µM. Here, we showed that TMMC induces apoptosis in activated HSCs at higher concentrations (30-50 µM), but is not cytotoxic to primary hepatocytes. Moreover, TMMC induces hyperacetylation of histone by inhibiting histone deacetylase (HDAC) in activated HSCs. Interestingly, TMMC treatment remarkably increased Fas-ligand (FasL) mRNA expression in a dose-dependent manner. Cycloheximide treatment reversed the induction of TMMC on apoptosis, indicating that de novo protein synthesis was required for TMMC-induced apoptosis in activated HSCs. In addition, FasL synthesis by TMMC is closely associated with maximal procaspase-3 proteolytic processing. In vivo, TMMC reduced activated HSCs in CCl(4)-intoxicated rats during liver injury recovery, as demonstrated by α-smooth muscle actin expression in rat liver. TMMC treatment also resulted in apoptosis, as demonstrated by cleavage of poly(ADP-ribose) polymerase in rat liver. In conclusion, TMMC may have therapeutic potential by inducing HSC apoptosis for the treatment of hepatic fibrosis.
Asunto(s)
Apoptosis/efectos de los fármacos , Chalconas/farmacología , Proteína Ligando Fas/metabolismo , Células Estrelladas Hepáticas/citología , Células Estrelladas Hepáticas/metabolismo , Animales , Tetracloruro de Carbono/toxicidad , Epigénesis Genética/efectos de los fármacos , Proteína Ligando Fas/biosíntesis , Proteína Ligando Fas/genética , Células Estrelladas Hepáticas/efectos de los fármacos , Inhibidores de Histona Desacetilasas/farmacología , Histonas/metabolismo , Inyecciones , Hígado/citología , Hígado/efectos de los fármacos , Hígado/metabolismo , Masculino , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas , Ratas Sprague-Dawley , Transcripción Genética/efectos de los fármacosRESUMEN
Tanshinone IIA, a major component extracted from the traditional herbal medicine, Salvia miltiorrhiza Bunge, improves blood circulation and treats chronic hepatitis and hepatic fibrosis. Activation of hepatic stellate cells (HSCs) is the predominant event in liver fibrosis. The therapeutic goal in liver fibrosis is the reversal of fibrosis and selective clearance of activated HSCs. We used rat HSCs transformed by Simian virus 40 (t-HSC/Cl-6) to overcome the limitations inherent in studying subcultures of HSCs. Treatment of t-HSC/Cl-6 cells with tanshinone IIA inhibited cell viability in a dose- and time-dependent manner. Tanshinone IIA induced apoptosis as demonstrated by DNA fragmentation, poly(ADP-ribose) polymerase and caspase-3 cleavage, increased Bax/Bcl-2 protein ratio, and depolarization of mitochondrial membranes to facilitate cytochrome c release into the cytosol. Furthermore, this compound markedly induced S phase cell cycle arrest, and down-regulated cyclins A and E, and cdk2. Thus, tanshinone IIA induces apoptosis and S phase cell cycle arrest in rat HSCs in vitro.
Asunto(s)
Apoptosis/efectos de los fármacos , Células Estrelladas Hepáticas/efectos de los fármacos , Fenantrenos/farmacología , Fase S/efectos de los fármacos , Abietanos , Animales , Caspasa 3/metabolismo , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Mitocondrias Hepáticas/efectos de los fármacos , Mitocondrias Hepáticas/fisiología , Proteínas Quinasas Activadas por Mitógenos/fisiología , RatasRESUMEN
The present study to evaluate the potential of constituents of the bark of Alnus japonica as a functional food with medicinal properties led to the identification of one new diarylheptanoid, named alusenone (1A), and 11 known ones (1B and 2-11). Their antioxidative and hepatoprotective activities were accessed by, respectively, a TOSC assay and a TBH-induced hepatotoxicity rat model. Mixtures 1, 2-6, 10, and 11 showed good antioxidative and hepatoprotective effects as compared with the positive controls.
Asunto(s)
Alnus/química , Antioxidantes/química , Enfermedad Hepática Inducida por Sustancias y Drogas/prevención & control , Diarilheptanoides/farmacología , Corteza de la Planta/química , Animales , Antioxidantes/farmacología , Diarilheptanoides/química , Estructura Molecular , RatasRESUMEN
Salvia miltiorrhiza is traditionally used to treat liver disease in Asia. In this study, we tested the ability of a purified extract of S. miltiorrhiza (PF2401-SF) and its constituents, tanshinone I, tanshinone IIA, and cryptotanshinone, to protect against acute and subacute liver damage induced by carbon tetrachloride by measuring serum transaminase levels, the reduced form of glutathione (GSH), antioxidant enzyme activities, and lipid peroxidation levels in the liver. We also evaluated their ability to protect primary cultured rat hepatocytes from tertiary-butylhydroperoxide (tBH) or d-galactosamine (GalN). PF2401-SF was protective at 50-200mg/kg per day in acute liver injury and 25-100mg/kg per day in subacute liver injury. Tanshinone I, tanshinone IIA, and cryptotanshinon (40 microM), inhibited lactate dehydrogenase leakage, GSH depletion, lipid peroxidation and free radical generation in vitro. PF2401-SF and its major constituents, tanshinone I, tanshinone IIA and cryptotanshinone, can protect against liver toxicity in vivo and in vitro due to its antioxidant effects.
Asunto(s)
Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Hepatocitos/efectos de los fármacos , Fenantrenos/farmacología , Extractos Vegetales/farmacología , Salvia miltiorrhiza/química , Abietanos , Animales , Antioxidantes/farmacología , Tetracloruro de Carbono/toxicidad , Relación Dosis-Respuesta a Droga , Masculino , Fenantrenos/química , Extractos Vegetales/química , Ratas , Ratas Sprague-DawleyRESUMEN
In this study, we investigated the hepatoprotective effects of four compounds from Galla Rhois [gallic acid methyl ester, gallic acid, an equilibrium mixture of 3-galloyl-gallic acid and 4-galloyl-gallic acid isomers, and 1,2,3,4,6-penta- O-galloyl- beta- D-glucose (PGG)] in primary rat hepatocytes undergoing necrosis or apoptosis. Treatment with gallic acid methyl ester (12.5 and 50 microM) or PGG (3.125, 12.5 and 50 microM) reduced hepatocyte necrosis induced by tert-butyl hydroperoxide. PGG treatment (4 and 20 microM) also altered hepatocyte apoptosis induced by glycochenodeoxycholic acid. Based on these results, we propose that PGG warrants further evaluation as a hepatoprotective agent, because it protected primary rat hepatocytes from both necrosis and apoptosis.
Asunto(s)
Apoptosis/efectos de los fármacos , Hepatocitos/efectos de los fármacos , Taninos Hidrolizables/farmacología , Animales , Áfidos , Células Cultivadas , Ácido Gálico/análogos & derivados , Ácido Gálico/aislamiento & purificación , Ácido Gálico/farmacología , Ácido Gálico/uso terapéutico , Taninos Hidrolizables/aislamiento & purificación , Taninos Hidrolizables/uso terapéutico , Necrosis/prevención & control , Tumores de Planta , Ratas , Rhus/químicaRESUMEN
Bile acid-induced hepatocyte apoptosis plays an important role in cholestatic liver disease, and the role of apoptosis may be of therapeutic interest in preventing liver disease. The dried root of Salvia miltiorrhiza Bunge (Labiatae) has been used traditionally to treat liver diseases. We investigated the antiapoptotic effects of a standardized fraction of S. miltiorrhiza (PF2401-SF) and its components, tanshinone I, tanshinone IIA, and cryptotanshinone, in primary cultured rat hepatocytes. PF2401-SF was enriched with tanshinone I (11.5%), tanshinone IIA (41.0%), and cryptotanshinone (19.1%). Glycochenodeoxycholic acid (GCDC)-induced apoptosis, as shown by DNA fragmentation, poly(ADP-ribose) polymerase cleavage, and activation of caspases-8, -9, and -3. PF2401-SF and its components, tanshinone I, tanshinone IIA, and cryptotanshinone showed antiapoptotic activity. Treatment with PF2401-SF or with its components significantly inhibited the generation of intracellular reactive oxygen species. Hydrophobic bile acids activate c-Jun-NH(2)-terminal kinase (JNK), p38 mitogen-activated protein kinases (MAPK), and extracellular signal-regulated kinase 1/2, and PF2401-SF inhibited the phosphorylation of JNK and p38. All three components of PF2401-SF inhibited JNK phosphorylation. Addition of inhibitors of MAPK showed that inhibition of JNK decreased apoptosis. These data indicate that PF2401-SF and its components protect hepatocytes from GCDC-induced apoptosis in vitro by inhibiting JNK.
Asunto(s)
Apoptosis/efectos de los fármacos , Ácidos y Sales Biliares/antagonistas & inhibidores , Ácidos y Sales Biliares/toxicidad , Hepatocitos/efectos de los fármacos , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Fenantrenos/farmacología , Salvia/química , Abietanos , Animales , Western Blotting , Caspasas/metabolismo , Células Cultivadas , Colagogos y Coleréticos/antagonistas & inhibidores , Colagogos y Coleréticos/toxicidad , Cromatografía Líquida de Alta Presión , Colorimetría , Fragmentación del ADN/efectos de los fármacos , Activación Enzimática/efectos de los fármacos , Ácido Glicoquenodesoxicólico/antagonistas & inhibidores , Ácido Glicoquenodesoxicólico/toxicidad , Estrés Oxidativo/efectos de los fármacos , Fosforilación/efectos de los fármacos , Extractos Vegetales/farmacología , Ratas , Especies Reactivas de Oxígeno/metabolismoRESUMEN
We investigated the protective effect of butein on glycochenodeoxycholic acid (GCDC)-induced apoptosis in primary cultured rat hepatocytes. Treatment with GCDC at a concentration of 100 microM for 4 h induced apoptosis, and treatment with butein at concentrations of 30 microM inhibited the GCDC-induced apoptosis as shown by the reduced cleavage of poly(ADP-ribose) polymerase, DNA fragmentation, and activation of caspases-3, -8, and -9. c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) play fundamental roles in cell survival, proliferation, and apoptosis. GCDC alone induced ERK and JNK phosphorylation. Butein alone induced ERK activation, and ERK activation was greater in hepatocytes treated with butein and GCDC than in hepatocytes exposed to GCDC alone. Butein treatment reduced JNK activation induced by GCDC. Addition of U0126, an inhibitor of ERK, did not alter the proapoptotic effect of GCDC or the antiapoptotic effect of butein. Addition of SP600125, a specific JNK inhibitor, protected hepatocytes against GCDC-induced apoptosis. These data suggest that butein has a protective effect against GCDC-induced hepatocyte apoptosis and that the protective effect of butein is JNK dependent but ERK independent.
Asunto(s)
Apoptosis/efectos de los fármacos , Hepatocitos/efectos de los fármacos , Fitoterapia , Extractos Vegetales/farmacología , Rhus , Ácidos y Sales Biliares , Chalconas/administración & dosificación , Chalconas/farmacología , Chalconas/uso terapéutico , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Hepatocitos/metabolismo , Humanos , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Extractos Vegetales/administración & dosificación , Extractos Vegetales/uso terapéuticoRESUMEN
Liver fibrosis and cirrhosis may be reversible, possibly through the selective clearance of activated hepatic stellate cells/myofibroblasts by apoptosis. Hepatic stellate cells transdifferentiate into myofibroblast-phenotype cells in culture, a process that recapitulates hepatic stellate cell activation in vivo. Bakuchiol, a prenylated phenolic terpene isolated from the seed of Psoralea corylifolia L. (Leguminosae), reduced activated hepatic stellate cells when treated to rats during liver injury recovery period as demonstrated by alpha-smooth muscle actin immunostaining in rat liver and induced apoptosis in activated hepatic stellate cells/myofibroblasts as demonstrated by DNA fragmentation, activation of caspase-3, release of cytochrome c into the cytoplasm, translocation of Bax into mitochondria, and the proteolytic cleavage of poly(ADP-ribose) polymerase (PARP) in vitro. Bakuchiol-induced apoptosis was prevented by z-DEVD-fmk, a specific inhibitor of caspase-3, and z-VAD-fmk, a general caspase inhibitor, suggesting that bakuchiol-induced apoptosis occurs through a caspase-3-dependent pathway in vitro. Bakuchiol treatment stimulated the activation of extracellular signal-regulated kinase 1/2 (ERK), c-Jun NH2-terminal protein kinase (JNK), and p38 mitogen-activated protein kinases (MAPK) in vitro. Pretreatment with SP600125 attenuated the bakuchiol-induced translocation of Bax into mitochondria, cytochrome c release into the cytosol, caspase-3 activation, and PARP cleavage. In contrast, preincubation with SB203580, a p38 MAPK inhibitor, and U0126, an ERK inhibitor, had no effect on bakuchiol-induced cell death and caspase-3 activity. Taken together, these findings indicate that bakuchiol induces caspase-3-dependent apoptosis through the activation of JNK, followed by Bax translocation into mitochondria in rat liver myofibroblasts.
Asunto(s)
Apoptosis/efectos de los fármacos , Caspasa 3/metabolismo , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Hígado/efectos de los fármacos , Fenoles/farmacología , Sustancias Protectoras/farmacología , Proteína X Asociada a bcl-2/metabolismo , Actinas/metabolismo , Animales , Tetracloruro de Carbono , Células Cultivadas , Enfermedad Hepática Inducida por Sustancias y Drogas , Citocromos c/metabolismo , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Activación Enzimática/efectos de los fármacos , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Hígado/metabolismo , Hígado/patología , Hepatopatías/metabolismo , Hepatopatías/prevención & control , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Masculino , Mitocondrias Hepáticas/efectos de los fármacos , Mitocondrias Hepáticas/metabolismo , Transporte de Proteínas/efectos de los fármacos , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Ratas , Ratas Sprague-Dawley , Factores de TiempoRESUMEN
We investigated the apoptotic effects of the protopanaxadiol ginsenosides, Rb (1) and Rb (2), and their intestinal bacterial metabolite, 20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol (M1), and of the protopanaxatriol ginsenoside, Rg (1), and its intestinal bacterial metabolite, 20(S)-protopanaxatriol, in activated rat hepatic stellate cells (HSCs) transformed by Simian virus 40 (T-HSC/Cl-6). As HSCs play a central role in liver fibrosis, agents that selectively induce apoptosis of HSCs could be used to treat this disease. Apoptosis was measured using cell viability tests, DNA fragmentation analysis, and immunoblot analysis of poly(ADP-ribose) polymerase cleavage. M1 (40 microM for 24 h) significantly induced apoptosis in activated rat HSCs. M1 induced apoptosis in a dose-dependent manner as shown by DNA fragmentation, an increased population of cells in the sub-G1 phase, and reduced mitochondrial transmembrane potential. M1 induced caspase-3 activity in a dose- and time-dependent manner. A specific inhibitor of caspase-3 prevented induction of apoptosis by M1 as shown by DNA fragmentation analysis. It is concluded that M1 induces apoptosis in T-HSC/Cl-6 cells via caspase-3 activation.
Asunto(s)
Apoptosis/efectos de los fármacos , Caspasa 3/metabolismo , Ginsenósidos/farmacología , Hepatocitos/efectos de los fármacos , Animales , Línea Celular Transformada , Activación Enzimática/efectos de los fármacos , Ginsenósidos/química , Ginsenósidos/aislamiento & purificación , Hepatocitos/enzimología , RatasRESUMEN
Polyozellin, isolated from Polyozellus multiplex (Thelephoraceae), was investigated for its anti-inflammatory activity in the murine macrophage cell line RAW 264.7. Polyozellin inhibited both lipopolysaccharide (LPS)-induced nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) gene expression in a dose-dependent manner. The effects of polyozellin on the activation of nuclear factor-kappaB (NF-kappaB) and mitogen-activated protein (MAP) kinases in these cells were studied in order to elucidate the underlying mechanism. Polyozellin suppressed the activation of both LPS-induced NF-kappaB and the stress-activated protein kinase (SAPK)/c-Jun N-terminal kinase (JNK), but had no effect on the extracellular signal-regulated kinase (ERK) or p38. These data suggest that polyozellin suppresses iNOS expression by inhibiting the activation of NF-kappaB and SAPK/JNK, leading to the inhibition of NO production.
Asunto(s)
Antiinflamatorios/farmacología , Furanos/farmacología , Proteínas Quinasas JNK Activadas por Mitógenos/antagonistas & inhibidores , FN-kappa B/antagonistas & inhibidores , Óxido Nítrico Sintasa de Tipo II/antagonistas & inhibidores , Óxido Nítrico/biosíntesis , Animales , Antiinflamatorios/química , Antiinflamatorios/aislamiento & purificación , Basidiomycota/química , Línea Celular , Regulación hacia Abajo , Quinasas MAP Reguladas por Señal Extracelular/genética , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Furanos/química , Furanos/aislamiento & purificación , Proteínas Quinasas JNK Activadas por Mitógenos/genética , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Lipopolisacáridos/antagonistas & inhibidores , Lipopolisacáridos/metabolismo , Ratones , FN-kappa B/genética , FN-kappa B/metabolismo , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos/genética , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
Hydrophobic bile acid-induced apoptosis plays an important role in cholestatic liver disease, and its prevention may be of therapeutic interest. The aim of this study was to investigate the protective effect of honokiol on glycochenodeoxycholic acid-induced apoptosis in primary cultured rat hepatocytes. Glycochenodeoxycholic acid is a hydrophobic bile salt that accumulates intrahepatically during cholestasis and induces hepatocyte apoptosis at pathophysiological concentrations. Primary rat hepatocytes were pretreated with honokiol at concentrations of 40, 20 and 10 microM 5 min before glycochenodeoxycholic acid treatment. Incubation of hepatocytes with glycochenodeoxycholic acid at a concentration of 100 microM for 4 h induced apoptosis as shown by DNA fragmentation, chromatin condensation and cleavage of poly(ADP-ribose) polymerase. Pretreatment with honokiol at concentrations of 40, 20 and 10 microM significantly inhibited the generation of intracellular reactive oxygen species and reduced activation of caspases-8, -9, and -3 and cleavage of poly-(ADP-ribose) polymerase. Glycochenodeoxycholic acid treatment up-regulated phosphorylation of stress-activated protein kinase/c-jun-NH2-terminal kinase which was inhibited by honokiol treatment. Inhibition of stress-activated protein kinase/c-jun-NH2-terminal kinase phosphorylation by SP600125 protected hepatocytes from apoptosis induced by glycochenodeoxycholic acid. These data indicate that honokiol protects hepatocytes from apoptosis induced by glycochenodeoxycholic acid in vitro and this protection may be due to reduced oxidative stress and inhibition of stress-activated protein kinase/c-jun-NH2-terminal kinase phosphorylation.
Asunto(s)
Apoptosis/efectos de los fármacos , Compuestos de Bifenilo/farmacología , Hepatocitos/efectos de los fármacos , Proteínas Quinasas JNK Activadas por Mitógenos/efectos de los fármacos , Lignanos/farmacología , Estrés Oxidativo/efectos de los fármacos , Animales , Compuestos de Bifenilo/aislamiento & purificación , Caspasas/efectos de los fármacos , Células Cultivadas , Ácido Glicoquenodesoxicólico , Lignanos/aislamiento & purificación , Magnolia/química , Fosforilación/efectos de los fármacos , Corteza de la Planta/química , Poli(ADP-Ribosa) Polimerasas/efectos de los fármacos , RatasRESUMEN
The therapeutic goal in liver fibrosis is the reversal of fibrosis and the selective clearance by apoptosis of hepatic stellate cells (HSCs), which play a central role in liver fibrogenesis. In this study, the apoptotic effect of wogonin, oroxylin A, 2',5,6',7-tetrahydroxyflavone, skullcapflavone I, and baicalein, isolated from the dried root of Scutellaria baicalensis, was investigated in activated rat HSCs, T-HSC/Cl-6 cells transformed with the Simian virus 40. Among the isolated compounds, skullcapflavone I (20 microM for 24 h) significantly induced apoptosis in activated rat HSCs while there was no change in the cell viability of hepatocytes. Skullcapflavone I increased caspase-3 and -9 activities accompanied by the proteolytic cleavage of poly(ADP-ribose) polymerase. Specific inhibitors of caspase-3 and caspase-9 prevented the apoptotic process induced by skullcapflavone I. From these results, skullcapflavone I from S. baicalensis selectively induced apoptosis in T-HSC/Cl-6 cells via caspase-3 and caspase-9 activation.
Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Apoptosis/efectos de los fármacos , Flavonas/farmacología , Hígado/efectos de los fármacos , Fitoterapia , Extractos Vegetales/farmacología , Scutellaria baicalensis , Animales , Antineoplásicos Fitogénicos/administración & dosificación , Antineoplásicos Fitogénicos/uso terapéutico , Caspasas/metabolismo , Células Cultivadas , Flavonas/administración & dosificación , Flavonas/uso terapéutico , Citometría de Flujo , Hígado/citología , Hígado/enzimología , Cirrosis Hepática/tratamiento farmacológico , Masculino , Extractos Vegetales/administración & dosificación , Extractos Vegetales/uso terapéutico , Raíces de Plantas , Ratas , Ratas Sprague-DawleyRESUMEN
The aim of this study was to investigate the protective effect of (S)-bakuchiol isolated from the seed of Psoralea corylifolia, on liver injury. Primary rat hepatocyte intoxication was induced by tert-butyl hydroperoxide (tBH), carbon tetrachloride (CCl4) or D-galactosamine (D-GalN). Liver injury was induced by either CCl4 or D-GalN in rats. In vitro, the cellular leakage of lactate dehydrogenase and cell viability following treatment with hepatotoxicants were significantly improved by bakuchiol treatment at a concentration range of 25-200 microM for tBH, 100-200 microM for CCl4 and 100-200 microM for D-GalN-induced hepatocyte injury. Treatment with bakuchiol significantly inhibited lipid peroxidation and intracellular glutathione depletion in hepatocytes induced by tBH, CCl4 or D-GalN. Treatment with bakuchiol (25 or 50 mg/kg, p.o.) at 1, 24 and 48 h after subcutaneous injection of CCl4 significantly reduced the levels of aspartate transaminase and alanine transaminase in serum. Histological observations revealed that fatty acid changes, hepatocyte necrosis and inflammatory cell infiltration in CCl4-injured liver was improved when treated with bakuchiol. Bakuchiol treatment (25 and 50 mg/kg, p.o.) also significantly reduced the levels of aspartate transaminase and alanine transaminase in an acute liver injury model induced by D-GalN. From these results, bakuchiol has a protective effect against tBH, CCl4 or D-GalN-induced hepatotoxicity in vitro or in vivo.
Asunto(s)
Enfermedad Hepática Inducida por Sustancias y Drogas/prevención & control , Fenoles/farmacología , Fitoterapia , Sustancias Protectoras/farmacología , Psoralea , Alanina Transaminasa/sangre , Animales , Aspartato Aminotransferasas/sangre , Tetracloruro de Carbono , Enfermedad Hepática Inducida por Sustancias y Drogas/etiología , Relación Dosis-Respuesta a Droga , Galactosamina , Hígado/efectos de los fármacos , Hígado/enzimología , Masculino , Fenoles/administración & dosificación , Fenoles/uso terapéutico , Extractos Vegetales/administración & dosificación , Extractos Vegetales/farmacología , Extractos Vegetales/uso terapéutico , Sustancias Protectoras/administración & dosificación , Sustancias Protectoras/uso terapéutico , Ratas , Ratas Sprague-Dawley , Semillas , terc-ButilhidroperóxidoRESUMEN
The aim of our study was to clarify the apoptosis pathway induced by aloe emodin, an hydroxyanthraquinone present in aloe vera leaves, in rat hepatic stellate cells transformed by simian virus 40 (t-HSC/Cl-6), which retain the features of activated rat stellate cells. Apoptosis was determined by DNA fragmentation, caspase activity assay and western blotting analysis. Treatment of t-HSC/Cl-6 cells with 12.5, 25, or 50 microM aloe emodin inhibited t-HSC/Cl-6 cell viability in a dose- and time-dependent manner. The induction of apoptosis by aloe emodin was confirmed by typical DNA ladder formation and annexin v-propidium iodide flow-cytometric analysis. Aloe emodin treatment of t-HSC/Cl-6 cells caused activation of caspase-3 and caspase-9, detected with a caspase activity assay, although no change was observed in caspase-8 activity. Western blotting showed caspase-3 and caspase-9 active forms and the subsequent proteolytic cleavage of poly(ADP-ribose) polymerase. Aloe emodin induced mitochondrial membrane depolarization. Our data also show that cytochrome c increased in the cytosol but decreased in the mitochondria in a time-dependent manner. Increased Bax and unchanged Bcl-2 levels resulted in an increased Bax/Bcl-2 ratio. Thus, our research provides evidence that aloe emodin-induced apoptosis involves a mitochondria-associated apoptosis pathway.
Asunto(s)
Apoptosis , Emodina/farmacología , Mitocondrias/efectos de los fármacos , Animales , Antraquinonas , Caspasa 3 , Caspasa 9 , Caspasas/metabolismo , Línea Celular , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Citocromos c/metabolismo , Fragmentación del ADN , Potenciales de la Membrana/efectos de los fármacos , Mitocondrias/metabolismo , Mitocondrias/fisiología , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Ratas , Proteína X Asociada a bcl-2RESUMEN
The therapeutic goal in liver fibrosis is the reversal of fibrosis and the selective clearance of activated hepatic stellate cells (HSCs) by inducing apoptosis. Over the past several years, we have screened for natural products that mediate apoptosis in activated HSCs. Among the candidate compounds, honokiol, isolated from Magnoliae cortex, was found to induce apoptotic death in activated rat HSCs, while there was no cell viability change in hepatocytes, at concentrations of 12.5-50 microM. Apoptosis was identified by DNA fragmentation, activation of caspase-3 and -9, and the proteolytic cleavage of poly(ADP-ribose) polymerase, down-regulation of bcl-2 and the release of mitochondrial cytochrome c into the cytoplasm.