RESUMEN
Excessive alcohol use often results in alcoholic liver disease (ALD). An early change in the liver due to excessive drinking is hepatic steatosis, which may ultimately progress to hepatitis, liver fibrosis, cirrhosis, and liver cancer. Among these debilitating processes, hepatic steatosis is reversible with the appropriate treatment. Therefore, it is important to find treatments and foods that reverse hepatic steatosis. Black carrot has antioxidant and anti-inflammatory effects. In this study, we examined the effectiveness of black carrot extract (BCE) on hepatic steatosis in in vivo and in vitro ethanol-induced liver injury models. For the in vivo experiments, serum aminotransferase activities enhanced by ethanol- and carbon tetrachloride were significantly suppressed by the BCE diet. Furthermore, morphological changes in the liver hepatic steatosis and fibrosis were observed in the in vivo ethanol-induced liver injury model, however, BCE feeding resulted in the recovery to an almost normal liver morphology. In the in vitro experiments, ethanol treatment induced reactive oxygen species (ROS) levels in hepatocytes at 9 h. Conversely, ROS production was suppressed to control levels and hepatic steatosis was suppressed when hepatocyte culture with ethanol were treated with BCE. Furthermore, we investigated enzyme activities, enzyme protein levels, and messenger RNA levels of alcohol dehydrogenase (ADH), cytochrome p450 2E1 (CYP2E1), and aldehyde dehydrogenase (ALDH) using enzyme assays, western blot, and quantitative reverse transcription-polymerase chain reaction analyses. We found that the activities of ADH, CYP2E1, and ALDH were regulated through the cAMP-PKA pathway at different levels, namely, translational, posttranslational, and transcriptional levels, respectively. The most interesting finding of this study is that BCE increases cAMP levels by suppressing the Pde4b mRNA and PDE4b protein levels in ethanol-treated hepatocytes, suggesting that BCE may prevent ALD.
Asunto(s)
Enfermedad Hepática Crónica Inducida por Sustancias y Drogas , Daucus carota , Hígado Graso , Hepatopatías Alcohólicas , Etanol/metabolismo , Citocromo P-450 CYP2E1/genética , Citocromo P-450 CYP2E1/metabolismo , Fosfodiesterasas de Nucleótidos Cíclicos Tipo 4/metabolismo , Fosfodiesterasas de Nucleótidos Cíclicos Tipo 4/farmacología , Especies Reactivas de Oxígeno/metabolismo , Daucus carota/metabolismo , Alcohol Deshidrogenasa/genética , Alcohol Deshidrogenasa/metabolismo , Alcohol Deshidrogenasa/farmacología , Antioxidantes/farmacología , ARN Mensajero/metabolismo , Tetracloruro de Carbono , Hígado/metabolismo , Hígado Graso/metabolismo , Aldehído Deshidrogenasa/metabolismo , Aldehído Deshidrogenasa/farmacología , Cirrosis Hepática , Transaminasas/metabolismo , Antiinflamatorios/farmacologíaRESUMEN
We studied the epigenetic regulation of how black carrot extract (BCE) protects against ethanol-induced hepatic damage. We have shown that the butanol-extracted fraction of BCE (BCE-BuOH) increased intracellular cyclic adenosine monophosphate (cAMP) levels by suppressing the expression of phosphodiesterase 4b (PDE4b); however, the detailed mechanism remains to be elucidated. We focused on changes in histone modifications involved in the suppression of pde4 expression. The methylation level of histone H3 lysine 9 (H3K9), which regulates gene expression of PDE4b, decreased after treatment with 100 mM ethanol but was significantly increased by treatment with 400 µg/ml BCE-BuOH. In contrast, ethanol induced an increase in H3K9 acetylation. However, treatment with BCE-BuOH inhibited the increase in acetylation through an increase in Sirtuin 1 (Sirt1), a histone deacetylase. Furthermore, BCE-BuOH treatment increased the level of methionine adenosyltransferase (MAT) 2a mRNA and increased intracellular S-adenosylmethionine. The present results indicate that BCE-BuOH is useful for protection against alcohol-induced hepatic injury. PRACTICAL APPLICATIONS: We have reported that black carrot extract (BCE) suppressed liver steatosis and liver fibrosis on a rat alcoholic liver disease model. The results from this study have shown that BCE regulated the alcoholic-induced hepatic injury at the level of epigenetic modifications. These results suggested that BCE is useful for protection against alcoholic-induced hepatic injury.
Asunto(s)
Daucus carota , Epigénesis Genética , Adenosina Monofosfato , Animales , Butanoles , Fosfodiesterasas de Nucleótidos Cíclicos Tipo 4/metabolismo , Daucus carota/genética , Etanol , Histonas/metabolismo , Lisina/metabolismo , Metionina Adenosiltransferasa/metabolismo , Extractos Vegetales , ARN Mensajero , Ratas , S-Adenosilmetionina/metabolismo , Sirtuina 1/genética , Sirtuina 1/metabolismoRESUMEN
1'-Acetoxychavicol acetate (ACA) is found in the rhizomes or seeds of Alpinia galanga and Alpinia conchigera, which are used as traditional spices in cooking and traditional medicines in Southeast Asia. ACA possesses numerous medicinal properties. Those include anticancer, antiobesity, antiallergy, antimicrobial, antidiabetic, gastroprotective, and anti-inflammatory activities. ACA is also observed to exhibit antidementia activity. Recent studies have demonstrated that combining ACA with other substances results in synergistic anticancer effects. The structural factors that regulate the activity of ACA include (1) the acetyl group at position 1', (2) the acetyl group at position 4, and (3) the unsaturated double bond between positions 2' and 3'. ACA induces the activation of AMP-activated protein kinase (AMPK), which regulates the signal transduction pathways, and has an important role in the prevention of diseases, including cancer, obesity, hyperlipidemia, diabetes, and neurodegenerative disorders. Such findings suggest that AMPK has a central role in different pharmacological functions of ACA, and ACA is useful for the prevention of life-threatening diseases. However, more studies should be performed to evaluate the clinical effects of ACA and to better understand its potential.
Asunto(s)
Alpinia/química , Alcoholes Bencílicos/farmacología , Extractos Vegetales/farmacología , Antiinfecciosos , Alcoholes Bencílicos/química , Demencia/tratamiento farmacológico , Humanos , Neoplasias/tratamiento farmacológico , Obesidad/tratamiento farmacológico , Rizoma/químicaRESUMEN
Protein degradation systems are critical pathways for the maintenance of protein homeostasis. The age-dependent attenuation of the proteasome activity contributes to age-related neurodegenerative processes. The molecule 1'-acetoxychavicol acetate (ACA) is naturally obtained from the rhizomes and seeds of Zingiberaceae plants, such as Languas galangal and Alpinia galangal, and exhibits anti-carcinogenic effects. Recently, we have shown that ACA protected the age-related learning and memory impairments in senescence-accelerated mice and maintained cognitive performance. Therefore, we here examined the effects of ACA on the protein degradation systems and cell protection against neurotoxicity in differentiated PC12 cells. ACA increased proteasome activity in PC12 cells. Increased proteasome activity occurred during the initial stages of ACA treatment and lasted at least 9 h. The activity returned to control levels within 24 h. The increase in proteasome activity by ACA was suppressed by H-89, which is a cAMP-dependent protein kinase A inhibitor. ACA increased the adenylate cyclase activity and therefore the intracellular cAMP levels. Furthermore, ACA recovered the initial cell viability, which was reduced after the addition of the amyloid ß-protein fragment to neuronally differentiated PC12 cells. The effects of ACA on amyloid toxicity were reduced after treatment with MG132, a proteasome inhibitor. These results demonstrated a neuroprotective effect of ACA via activation of cAMP/cAMP-dependent protein kinase A signaling in neuronally differentiated PC12 cells.
Asunto(s)
Alcoholes Bencílicos/farmacología , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Complejo de la Endopetidasa Proteasomal/efectos de los fármacos , Animales , Fármacos Neuroprotectores/farmacología , Células PC12 , Proteolisis/efectos de los fármacos , Ratas , Transducción de SeñalRESUMEN
1'-Acetoxychavicol acetate (ACA) is naturally obtained from the rhizomes and seeds of Alpinia galangal. Here, we examined the effect of ACA on learning and memory in senescence-accelerated mice prone 8 (SAMP8). In mice that were fed a control diet containing 0.02% ACA for 25 weeks, the learning ability in the Morris water maze test was significantly enhanced in comparison with mice that were fed the control diet alone. In the Y-maze test, SAMP8 mice showed decreased spontaneous alterations in comparison with senescence-accelerated resistant/1 (SAMR1) mice, a homologous control, which was improved by ACA pretreatment. Serum metabolite profiles were obtained by GC-MS analysis, and each metabolic profile was plotted on a 3D score plot. Based upon the diagram, it can be seen that the distribution areas for the three groups were completely separate. Furthermore, the contents of ß-hydroxybutyric acid and palmitic acid in the serum of SAMP8-ACA mice were higher than those of SAMP8-control mice and SAMR1-control mice. We also found that SAMR1 mice did not show histological abnormalities, whereas histological damage in the CA1 region of the hippocampus in SAMP8-control mice was observed. However, SAMP8-ACA mice were observed in a similar manner as SAMR1 mice. These findings confirm that ACA increases the serum concentrations of ß-hydroxybutyric acid and palmitic acid levels and thus these fuels might contribute to the maintenance of the cognitive performance of SAMP8 mice.
Asunto(s)
Envejecimiento/fisiología , Alcoholes Bencílicos/farmacología , Metabolismo Energético/efectos de los fármacos , Cuerpos Cetónicos/biosíntesis , Memoria Espacial/efectos de los fármacos , Ácido 3-Hidroxibutírico/sangre , Envejecimiento/efectos de los fármacos , Alpinia/química , Animales , Hipocampo/metabolismo , Aprendizaje por Laberinto , Metaboloma , Ratones , Neuronas , Ácido Palmítico/sangreRESUMEN
Alpinia galanga and Languas galanga, which are plants belonging to the ginger family, are frequently used for cooking, especially in Thai and Indonesian cuisine. The compound 1'-acetoxychavicol acetate (ACA), which is naturally obtained from the rhizomes and seeds of these gingers, has antioxidant and anti-inflammatory properties. We investigated the anti-obesity effects of ACA in 3T3-L1 adipocytes and in high fat diet (HFD)-induced rat models of obesity. ACA caused a significant decrease in the activity of GPDH in 3T3-L1 adipocytes without eliciting cell cytotoxicity, and it inhibited cellular lipid accumulation through the down-regulation of transcription factors such as PPARγ and C/EBPα. ACA also induced a dose-dependent phosphorylation of AMP-activated protein kinase (AMPK). In the animal model, rats fed an HFD containing 0.05% ACA gained less weight than rats fed an HFD alone. The visceral fat mass in rats fed an HFD containing 0.05% ACA tended to be lower than that in rats fed an HFD alone. Furthermore, a histological examination of livers from rats fed an HFD showed steatohepatitis. However, rats fed an HFD containing 0.05% ACA showed no histopathological changes in the liver tissue. Our results show that ACA exerts anti-obesity activities both in vitro and in vivo and suggests that ACA may have a novel preventive activity against obesity and possibly other metabolic diseases.
Asunto(s)
Adipocitos/efectos de los fármacos , Adipogénesis/efectos de los fármacos , Alcoholes Bencílicos/farmacología , Grasas de la Dieta/administración & dosificación , Células 3T3-L1 , Adenilato Quinasa/metabolismo , Adipocitos/metabolismo , Animales , Western Blotting , Masculino , Ratones , Fosforilación , Ratas , Ratas Sprague-DawleyRESUMEN
BACKGROUND: The development of alcoholic liver disease is a complex process that involves both the parenchymal and non-parenchymal cells of the liver. We examined the effect of an Ecklonia cava extract on ethanol-induced liver injury. METHODS: Isolated hepatocytes and hepatic stellate cells (HSCs) were incubated with ethanol. Ecklonia cava polyphenol (ECP) was added to the cultures that had been incubated with ethanol. Male Wistar rats were fed a diet that included 0.02% or 0.2% ECP or no ECP. For a period of 3 weeks, the animals were given drinking water containing 5% ethanol and were also treated with carbon tetrachloride (CCl4) (0.1 ml/kg of body weight). RESULTS: In the cultured hepatocytes, the ECP treatment suppressed the ethanol-induced increase in cell death by maintaining intracellular glutathione (GSH) levels. In HSCs, ECP treatment suppressed the ethanol-induced increases in type I collagen and α-smooth muscle actin expression by maintaining intracellular levels of reactive oxygen species and GSH. We examined the effects of ECP on serum AST and ALT activity, as well as the progression of liver fibrosis in rats treated with ethanol and CCl4. ECP treatment suppressed plasma AST and ALT activities in the ethanol- and CCl4-treated rats. ECP treatment fully protected the rats against ethanol- and CCl4-induced liver injury. GENERAL SIGNIFICANCE: ECP may be a candidate for preventing ethanol-induced liver injury.
Asunto(s)
Depresores del Sistema Nervioso Central/toxicidad , Enfermedad Hepática Inducida por Sustancias y Drogas/prevención & control , Etanol/toxicidad , Kava/química , Polifenoles/uso terapéutico , Sustancias Protectoras/uso terapéutico , Alanina Transaminasa/sangre , Animales , Aspartato Aminotransferasas/sangre , Western Blotting , Tetracloruro de Carbono/toxicidad , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Enfermedad Hepática Inducida por Sustancias y Drogas/etiología , Colágeno Tipo I/metabolismo , Glutatión/metabolismo , Células Estrelladas Hepáticas/citología , Células Estrelladas Hepáticas/efectos de los fármacos , Células Estrelladas Hepáticas/metabolismo , Hepatocitos/citología , Hepatocitos/efectos de los fármacos , Hepatocitos/metabolismo , Técnicas para Inmunoenzimas , Peroxidación de Lípido/efectos de los fármacos , Masculino , Ratas , Ratas Wistar , Especies Reactivas de Oxígeno/metabolismoRESUMEN
1,5-Anhydro-D-fructose (1,5-AF) is a monosaccharide that shares a structural similarity to glucose. 1,5-AF is found in fungi, algae, Escherichia coli and rat liver and is produced by the degradation of starch and glycogen, which is catalyzed by the enzyme alpha-1,4-glucan lyase. However, the physiological role of 1,5-AF in mammalian tissues is not well understood. Here, we investigated the anti-obesity potential of 1,5-AF on adipogenesis in 3T3-L1 adipocytes. 1,5-AF caused a significant decrease in GPDH activity in 3T3-L1 preadipocytes and mature adipocytes without eliciting cytotoxicity, and inhibited cellular lipid accumulation through down-regulation of transcription factors such as PPARgamma and C/EBPalpha. 1,5-AF also induced dose-dependent phosphorylation of AMP-activated protein kinase (AMPK), a cellular energy sensor. However, the total AMPK protein content remained unchanged. Furthermore, 1,5-AF increased the levels of reactive oxygen species, an important upstream signal for AMPK activation in 3T3-L1 adipocytes. Our results show that 1,5-AF exerts anti-obesity action in vitro and suggest that 1,5-AF is potentially a novel preventative agent for obesity and other metabolic diseases.
Asunto(s)
Adipocitos/efectos de los fármacos , Adipogénesis/efectos de los fármacos , Fármacos Antiobesidad/farmacología , Fructosa/análogos & derivados , Células 3T3 , Proteínas Quinasas Activadas por AMP/metabolismo , Adipocitos/metabolismo , Animales , Proteínas Potenciadoras de Unión a CCAAT/metabolismo , Evaluación Preclínica de Medicamentos , Fructosa/farmacología , Glicerolfosfato Deshidrogenasa/metabolismo , Ratones , PPAR gamma/metabolismoRESUMEN
(1'S)-acetoxychavicol acetate ((S)-ACA) exhibits chemopreventive effects on chemically induced tumor formation. It has been shown that ACA inhibited the development of azoxymethane-induced colon carcinogenesis through its suppression of cell proliferation in the colonic mucosa and its induction of glutathione S-transferase and quinone oxidoreductase 1 in vivo. In this study, we investigated how ACA induced these enzymes by using rat intestine epithelial cells (IEC6) in vitro. ACA induced glutathione S-transferase (GST) and NAD (P)H: quinone oxidoreductase 1 (NQO1) activities, increased intracellular glutathione (GSH) level, and upregulated intranuclear Nrf2 and cytosolic p21. It suggested that activation of phase II enzymes via Nrf2 associated with p21 is one of possible mechanisms of ACA to prevent advance of carcinogenesis.
Asunto(s)
Alpinia/química , Antineoplásicos Fitogénicos/farmacología , Alcoholes Bencílicos/farmacología , Neoplasias del Colon/enzimología , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/metabolismo , Activación Enzimática/efectos de los fármacos , Factor 2 Relacionado con NF-E2/metabolismo , Animales , Azoximetano , Línea Celular , Proliferación Celular/efectos de los fármacos , Colon/efectos de los fármacos , Colon/enzimología , Neoplasias del Colon/metabolismo , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Glutatión/metabolismo , Glutatión Transferasa/metabolismo , Mucosa Intestinal/efectos de los fármacos , Mucosa Intestinal/enzimología , Fase II de la Desintoxicación Metabólica/fisiología , NAD(P)H Deshidrogenasa (Quinona)/metabolismo , Extractos Vegetales/farmacología , RatasRESUMEN
Obesity, the leading metabolic disease in the world, is a serious health problem in industrialized countries. We investigated the anti-obesity effect of Blumea balsamifera extract on adipocyte differentiation of 3T3-L1 preadipocytes and anti-obesity effect of 3T3-L1 adipocytes. We found that treatment with an extract of Blumea balsamifera suppressed lipid accumulation and glycerol-3-phosphate dehydrogenase (GPDH) activity without affecting cell viability in 3T3-L1 preadipocytes and adipocytes. Furthermore, Blumea balsamifera extract brought significant attenuation of expressions of key adipogenic transcription factors, including peroxisome proliferator-activated receptor (PPAR)gamma, CCAAT element binding protein (C/EBPs) and leptin, however, induced up-regulation of adiponectin at the protein level in 3T3-L1 preadipocytes and adipocytes. These results suggest that Blumea balsamifera extract may block adipogenesis, at least in part, by decreasing key adipogenic transcription factors in 3T3-L1 preadipocytes and may have antiatherogenic, anti-inflammatory, and antidiabetic effects through up-regulation of adiponectin in 3T3-L1 adipocytes.
Asunto(s)
Adipocitos/efectos de los fármacos , Asteraceae/química , Extractos Vegetales/farmacología , Hojas de la Planta/química , Células 3T3-L1 , Adipocitos/citología , Adipocitos/metabolismo , Adipogénesis/efectos de los fármacos , Adiponectina/metabolismo , Animales , Compuestos Azo/química , Western Blotting , Proteína alfa Potenciadora de Unión a CCAAT/metabolismo , Proteína beta Potenciadora de Unión a CCAAT/metabolismo , Supervivencia Celular/efectos de los fármacos , Glicerolfosfato Deshidrogenasa/metabolismo , Leptina/metabolismo , Lípidos/análisis , Ratones , Obesidad/prevención & control , PPAR gamma/metabolismo , Coloración y Etiquetado/métodosRESUMEN
Obesity, the leading metabolic disease in the world, is a serious health problem in industrialized countries. The Zizyphus jujuba fruit has been used as traditional Chinese medicinal herb and considered to affect various physiological functions in the body for thousands of years. We investigated the anti-obesity effect of Z. jujuba on adipocyte differentiation of 3T3-L1 preadipocytes and found that treatment with an extract of Z. jujuba suppressed lipid accumulation and glycerol-3-phosphate dehydrogenase (GPDH) activity without affecting cell viability. Further fractionation of the initial Z .jujuba extract with organic solvent revealed that the chloroform fraction (CHCl(3)-F) elicited the most inhibitory effect, which involved significant attenuation of the expression of key adipogenic transcription factors, including peroxisome proliferator-activated receptor (PPAR)gamma and CCAAT enhancer binding proteins (C/EBPs) at the protein level. These results suggest that CHCl(3)-F may block adipogenesis, at least in part, by decreasing the expression of PPARgamma, C/EBPalpha and beta.
Asunto(s)
Adipocitos/efectos de los fármacos , Adipogénesis/efectos de los fármacos , Medicamentos Herbarios Chinos/farmacología , Células Madre/efectos de los fármacos , Ziziphus , Células 3T3-L1 , Adipocitos/citología , Adipocitos/metabolismo , Animales , Proteínas Potenciadoras de Unión a CCAAT/metabolismo , Diferenciación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Cloroformo/farmacología , Glicerolfosfato Deshidrogenasa/metabolismo , Ratones , Obesidad/tratamiento farmacológico , Obesidad/patología , PPAR gamma/metabolismo , Células Madre/citología , Triglicéridos/metabolismoRESUMEN
Hepatocellular carcinoma is a type of tumor highly resistant to available chemotherapeutic agents. The treatment of hepatocellular carcinoma remains a challenge that needs new approaches in the future. In a previous study, we demonstrated that the chloroform fraction (CHCl(3)-F) from Z. jujuba has anticancer activity in human liver cancer cells (HepG2), and that combining CHCl(3)-F with green tea extracts (GTE) results in enhanced effects of anticancer activity in the cells. To further understand the mechanism of the anticancer activity of combining CHCl(3)-F and GTE in HepG2 cells, we investigated whether the addition of a mixture of CHCl(3)-F and GTE would affect the expression of APRIL (a proliferation-inducing ligand), which was expressed in HepG2 cells from 4 hours of incubation in vitro. We have shown that CHCl(3)-F and GTE enhanced anti-cancer activity by reducing the expression of APRIL. We speculate that the CHCl(3)-F and GTE mixture might provide a lead to a new drug design to treat hepatocellular carcinoma in the future.
Asunto(s)
Antineoplásicos Fitogénicos/uso terapéutico , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Camellia sinensis , Carcinoma Hepatocelular/tratamiento farmacológico , Neoplasias Hepáticas/tratamiento farmacológico , Extractos Vegetales/uso terapéutico , Miembro 13 de la Superfamilia de Ligandos de Factores de Necrosis Tumoral/metabolismo , Ziziphus , Animales , Antineoplásicos Fitogénicos/farmacología , Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Carcinoma Hepatocelular/metabolismo , Ciclo Celular , Línea Celular Tumoral , Supervivencia Celular , Expresión Génica , Humanos , Fitoterapia , Extractos Vegetales/farmacología , Ratas , TéRESUMEN
Anticarcinogenic effects attributed to phytochemicals may be based on synergistic, additive, or antagonistic interactions of many compounds. In our previous study, we demonstrated that the chloroform fraction (CHCl(3)-F) from Z. jujuba has anticancer activity in HepG2 cells. In China, many people drink jujuba tea and believe in the synergic effects of jujuba and tea for better health. We therefore investigated the effects of CHCl(3)-F and green tea extract (GTE), and their underlying mechanisms of action in HepG2 cells. Our results showed that GTE enhanced the effect of CHCl(3)-F on cell viability in HepG2 cells, without cytotoxicity in rat hepatocytes, which was used as a normal cell model. Furthermore, combination of CHCl(3)-F and GTE caused an effect on G1 phase arrest but not on apoptosis. Interestingly, the mechanism of the G1 arrest was associated, not with an increase in p27(Kip1) levels and the hypohosphorylation of Rb, which are pathways used by CHCl(3)-F on G1 arrest in HepG2 cells, but with increases in p53 and p21(Waf1/Cip1) levels, and a decrease in cyclin E levels. Collectively, our findings suggest that combination of CHCl(3)-F and GTE produces an enhanced cell growth inhibition effect, and that the resultant G1 arrest was caused via a different mechanism as that of CHCl(3)-F treatment alone.
Asunto(s)
Apoptosis/efectos de los fármacos , Camellia sinensis , Carcinoma Hepatocelular/patología , Medicamentos Herbarios Chinos/farmacología , Neoplasias Hepáticas/patología , Té , Ziziphus , Animales , Carcinoma Hepatocelular/metabolismo , Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Células Cultivadas , Ciclina E/metabolismo , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/metabolismo , Inhibidor p27 de las Quinasas Dependientes de la Ciclina/metabolismo , ADN/metabolismo , Relación Dosis-Respuesta a Droga , Hepatocitos/efectos de los fármacos , Hepatocitos/metabolismo , Hepatocitos/patología , Humanos , Neoplasias Hepáticas/metabolismo , Masculino , Ratas , Ratas Wistar , Especies Reactivas de Oxígeno/metabolismo , Proteína de Retinoblastoma/metabolismo , Proteína p53 Supresora de Tumor/metabolismoRESUMEN
FK506 (tacrolimus) is a widely used immunosuppressant first employed in the management of rejection in organ transplantation, but now used for autoimmune disease. However, the nephrotoxicity induced by FK506 remains a serious clinical problem. We previously demonstrated that FK506 caused a significant increase in apoptosis of LLC-PK1 cells, a porcine proximal tubule cell line, but the addition of green tea extract and its polyphenolic components suppressed the cell death. Here, we examined the synergistic effect of tea polyphenols on the protection of FK506-induced cell death. The combined treatment with 5 microM (-)-epigallocatechin-gallate (EGCG) and 5 microM of (+)-catechin (C), (-)-epicatechin (EC), (-)-epigallocatechin (EGC) or (-)-epicatechin-gallate (ECG) reduced FK506-induced cytotoxicity in LLC-PK1. Similarly, the combined treatment with 5 microM EGC and 5 microM of C, EC, EGCG or ECG also reduced the cytotoxicity. These results showed that the co-treatments with EGCG and EGC, EGCG or ECG, and EGC and ECG have stronger synergistic effects on the protection of FK506-induced cell death. Furthermore, the combined treatment of EGCG (5 microM) and EGC (5 microM) showed a significant time-dependent suppression of the increased intracellular ROS levels 15 min after the addition of FK506, as well as on caspase activation. The results of these synergistic effects of the constituents of green tea extract suggest that its protective effects may reside in more than just one of its constituent.
Asunto(s)
Apoptosis/efectos de los fármacos , Bebidas , Flavonoides/farmacología , Inmunosupresores/farmacología , Túbulos Renales Proximales/citología , Fenoles/farmacología , Tacrolimus/farmacología , Animales , Caspasa 3/metabolismo , Catequina/análogos & derivados , Catequina/farmacología , Línea Celular , Supervivencia Celular/efectos de los fármacos , Sinergismo Farmacológico , Inmunosupresores/efectos adversos , Túbulos Renales Proximales/efectos de los fármacos , Túbulos Renales Proximales/metabolismo , Polifenoles , Especies Reactivas de Oxígeno/metabolismo , Porcinos , Tacrolimus/efectos adversosRESUMEN
Blumea balsamifera is known to improve physiological disorders such as rheumatism and hypertension, but its anticancer activity has not been well elucidated. In this study, we found that Blumea balsamifera MeOH extract (BME) induced growth-inhibitory activity in rat and human hepatocellular carcinoma cells without cytotoxicity in rat hepatocytes which were used as a normal cell model. BME induced cell cycle arrest at the G1 phase via decreases in the expression of cyclin-E and phosphorylation of retinoblastoma protein. Furthermore, BME reduced the level of a proliferation-inducing ligand, that stimulates tumor cell growth. These findings suggest that BME has possible therapeutic potential in hepatoma cancer patients and that depletion of cellular APRIL is an important mechanism in the growth-inhibitory effect of BME.
Asunto(s)
Asteraceae/química , Carcinoma Hepatocelular/patología , Inhibidores de Crecimiento/farmacología , Extractos Vegetales/farmacología , Animales , Carcinoma Hepatocelular/tratamiento farmacológico , Carcinoma Hepatocelular/genética , Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Ciclina E/metabolismo , Etanol/química , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Inhibidores de Crecimiento/uso terapéutico , Humanos , Metanol/química , Inhibidores de la Síntesis del Ácido Nucleico/farmacología , Inhibidores de la Síntesis del Ácido Nucleico/uso terapéutico , Fosforilación/efectos de los fármacos , Extractos Vegetales/uso terapéutico , Procesamiento Proteico-Postraduccional/efectos de los fármacos , Ratas , Retinoblastoma/metabolismo , Miembro 13 de la Superfamilia de Ligandos de Factores de Necrosis Tumoral/metabolismoRESUMEN
Blumea balsamifera (also known as sambong), a medicinal plant, is known to improve physiological disorders such as rheumatism and hypertension. However, its anticancer activity has not been well elucidated. In this study, we found that Blumea balsamifera MeOH extract (BME) induced growth inhibitory activity in rat and human hepatocellular carcinoma cells (McA-RH7777, HepG2, respectively) without cytotoxicity as in with rat hepatocytes used as a normal cell model. BME induced cell cycle arrest at G1 phase via decreases in expression of cyclin-E and phosphorylation of retinoblastoma (Rb) protein in both dose- and time-dependent manners. Furthermore, BME reduced the level of a proliferation related ligand (APRIL) which stimulates tumor cell growth. The anti-proliferative effect of BME was improved slightly but significantly by the treatment with recombinant human APRIL. These findings suggest that BME may have a possible therapeutic potential in hepatoma cancer patients and the depletion of cellular APRIL may be one of the important mechanisms on the growth inhibitory effect of BME.
Asunto(s)
Asteraceae/química , Carcinoma Hepatocelular/patología , Neoplasias Hepáticas/patología , Extractos Vegetales/farmacología , Animales , Ciclina E/metabolismo , Relación Dosis-Respuesta a Droga , Resistencia a Antineoplásicos , Fase G1/efectos de los fármacos , Humanos , Fosforilación/efectos de los fármacos , Ratas , Proteína de Retinoblastoma/metabolismo , Células Tumorales Cultivadas , Miembro 13 de la Superfamilia de Ligandos de Factores de Necrosis Tumoral/metabolismo , Miembro 13 de la Superfamilia de Ligandos de Factores de Necrosis Tumoral/fisiologíaRESUMEN
The Zizyphus jujuba fruit has been used as a traditional Chinese medicinal herb and considered to affect various physiological functions in the body for thousands of years. However, its anti-cancer activity and mechanism of action remain to be elucidated. We investigated the anti-cancer activity of Zizyphus jujuba Mill and its underlining mechanisms of action in human hepatoma cells (HepG2) and found that the extract of Z. jujuba decreased the viability of the cells. Further extraction of the initial Z. jujuba extract with organic solvents revealed that the chloroform fraction (CHCl(3)-F) was the most effective. Interestingly, the CHCl(3)-F induced not only apoptosis but also G1 arrest at a low concentration (100 mug/ml) and G2/M arrest at a higher concentration (200 mug/ml) by cell cycle assay. Apoptosis, an increase in intracellular ROS (reactive oxygen species) level, a decline of mitochondrial membrane potential at low Z. jujuba concentrations, and a ROS-independent mitochondrial dysfunction pathway at high concentrations were all observed. CHCl(3)-F-induced G1 arrest in HepG2 cells was associated with an increase in hypohosphorylation of Rb and p27(Kip1), and a decrease of phosphorylated Rb. However, CHCl(3)-F-induced G2/M arrest in HepG2 cells correlated with a decrease of the p27(Kip1) levels and generation of the phosphorylation of p27(Kip1), however the hypohosphorylation of Rb protein remained. Collectively, our findings suggest that the CHCl(3)-F extract of Z. jujuba extract induced a concentration dependent effect on apoptosis and a differential cell cycle arrest in HepG2 cells.
Asunto(s)
Antineoplásicos/farmacología , Medicamentos Herbarios Chinos/farmacología , Ziziphus , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Cloroformo , Inhibidor p27 de las Quinasas Dependientes de la Ciclina/metabolismo , Fase G1/efectos de los fármacos , Fase G2/efectos de los fármacos , Humanos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Fosforilación/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Proteína de Retinoblastoma/metabolismo , SolventesRESUMEN
The constituents of cape aloe were investigated after a preliminary screening of the growth-inhibiting effect on Ehrlich ascites tumor cells (EATC) of several extracts of this plant. Ten compounds were isolated from the dichloromethane (CH(2)Cl(2)) extract that showed the strongest activity, and their structures were elucidated as aloe-emodin (1), p-hydroxybenzaldehyde (2), p-hydroxyacetophenone (3), pyrocatechol (4), 10-oxooctadecanoic acid (5), 10-hydroxyoctadecanoic acid (6), methyl 10-hydroxyoctadecanoate (7), 7-hydroxy-2,5-dimethylchromone (8), furoaloesone (9), and 2-acetonyl-8-(2-furoylmethyl)-7-hydroxy-5-methylchromone (10) based on MS and various NMR spectroscopic techniques. Compounds 2-7 were isolated for the first time from cape aloe. Compounds 4-7 and 10 showed a significant growth-inhibiting effect, and compound 1 exhibited a remarkable synergistic effect on compounds 8-10, which was not observed with the treatment by each compound alone on EATC. These results suggest that the strong growth-inhibiting effect of the CH(2)Cl(2) extract was dependent not on one compound alone, but on the synergistic effect from the combination of compound 1 and the other compounds.
Asunto(s)
Aloe/química , Antineoplásicos/farmacología , Carcinoma de Ehrlich/tratamiento farmacológico , Fitoterapia , Acetofenonas/química , Acetofenonas/farmacología , Animales , Antraquinonas , Antineoplásicos/química , Benzaldehídos/química , Benzaldehídos/farmacología , Carcinoma de Ehrlich/metabolismo , Carcinoma de Ehrlich/patología , Catecoles/química , Catecoles/farmacología , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Cromonas/química , Cromonas/farmacología , Sinergismo Farmacológico , Emodina/química , Emodina/farmacología , Espectrometría de Masas , Cloruro de Metileno/química , Estructura Molecular , Resonancia Magnética Nuclear Biomolecular , Extractos Vegetales/farmacología , Extractos Vegetales/uso terapéutico , Polvos/química , Factores de TiempoRESUMEN
Cape aloe (Aloe ferox Miller) has been a herb well known for its cathartic properties and has also been used popularly as a health drink (juice, tea and tonic) in the United States and in Europe. Cape aloe extract also has been reported to possess several pharmacological effects, such as anti-inflammatory, anti-bacterial, anti-fungal and protective effect against liver injury. However, the investigations on an anti-tumor activity in cape aloe extract are very few and subsequent mechanisms have not been well elucidated. In this study, we examined the effect of the selective growth inhibitory activity of cape aloe extract and found that the cape aloe extract, especially the dichloromethane (CH(2)Cl(2)) extract, caused a dose-dependent growth inhibitory effect in Ehrlich ascites tumor cells (EATC), but not in mouse embryo fibroblast (NIH3T3) cells, which was used as a normal cell model. Furthermore, the CH(2)Cl(2) extract caused an accumulation of cells in the G1 phase and a decrease of cells in the S and G2/M phase of the cell cycle and inhibited DNA synthesis in a dose-dependent manner. In addition, other results suggest that cell cycle arrest and inhibition of proliferation in EATC by the CH(2)Cl(2 )extract are associated with decreased retinoblastoma protein (Rb) phosphorylation.
Asunto(s)
Aloe , Carcinoma de Ehrlich/patología , Inhibidores de Crecimiento/farmacología , Extractos Vegetales/farmacología , Animales , Ciclo Celular/efectos de los fármacos , División Celular/efectos de los fármacos , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , ADN/biosíntesis , Relación Dosis-Respuesta a Droga , Fase G1/efectos de los fármacos , Fase G2/efectos de los fármacos , Ratones , Células 3T3 NIH , Fosforilación/efectos de los fármacos , Proteína de Retinoblastoma/metabolismo , Fase S/efectos de los fármacosRESUMEN
The nephrotoxicity induced by the immunosuppressive drug FK506 (tacrolimus or fujimycin), limits its usefulness in widespread application, and the underlying mechanism has not been completely understood. The primary targets of FK506 in the kidney are the proximal tubular epithelial cells. In this study, the protection of green tea extract against FK506-induced cell death of LLC-PK1 cells was investigated. FK506 caused a significant decrease in survival of the cells, but the addition of green tea extract reduced this effect in a dose-dependent manner. Treatment of the cells with 50 microM (41.1 microg/ml) FK506 induced a significant increase in annexin V-positive/propidium iodide-negative cells from 2.68 to 14.5%, whereas the addition of 6.25, 12.5, and 25 microg/ml of green tea extract caused a significant protective effect in apoptotic cells from 14.5 to 6.51, 3.20 and 3.02%, respectively. The effect of five different constituent tea polyphenols was also examined. Epigallocatechin-gallate and epigallocatechin significantly reduced FK506-induced cytotoxicity but epicatechin and catechin had no effect on cell viability. Furthermore, changes in cytochrome c release and caspase activation, which characterize apoptosis, were studied. Epigallocatechin-gallate and epigallocatechin suppressed a significant release of cytochrome c and activation of caspase-3 in FK506-treated LLC-PK1 cells.