RESUMEN
Bioconversion of quercetin glucosides using four generally recognized as safe (GRAS) organisms (Aspergillus oryzae, Bacillus subtilis, Lactobacillus plantarum, and Saccharomyces cerevisiae) was evaluated by measuring changes in the levels of quercetin compounds of onion. Of the four organisms, S. cerevisiae increased the content of quercetin-3-O-ß-D-glucoside (III; isoquercitrin) and quercetin (IV), whereas decreasing quercetin-3,4'-O-ß-D-glucoside (I) and quercetin-4'-O-ß-D-glucoside (II). Also, S. cerevisiae converted authentic compound I to III, and II to IV, respectively. These results suggest that S. cerevisiae can be used to increase the levels of isoquercitrin (III), the most bioavailable quercetin compound in onion.
Asunto(s)
Glucósidos/metabolismo , Cebollas/química , Quercetina/metabolismo , Saccharomyces cerevisiae/metabolismo , Glucósidos/análisis , Glucósidos/química , Estructura Molecular , Quercetina/análogos & derivados , Quercetina/análisis , Quercetina/química , Estereoisomerismo , Regulación hacia ArribaRESUMEN
A new zymogram method, silver-stained fibrin zymography, for separation of protease bands and activity detection using a single substrate gel, was developed. The method takes advantage of the nanoscale sensitivity of both zymography and silver staining. After SDS-PAGE in a gel containing fibrin, the gel was incubated in enzyme reaction buffer and the zymogram was silver-stained. Bands with protease activity were stained with silver in clear areas where the protein substrate had been degraded. The molecular sizes of proteases were accurately determined. Furthermore, proteases of high molecular weight were clearly and sharply resolved.
Asunto(s)
Electroforesis en Gel de Poliacrilamida/métodos , Péptido Hidrolasas/aislamiento & purificación , Proteínas Bacterianas/química , Proteínas Bacterianas/aislamiento & purificación , Proteínas Bacterianas/metabolismo , Precursores Enzimáticos , Fibrina/química , Fibrina/metabolismo , Peso Molecular , Péptido Hidrolasas/química , Péptido Hidrolasas/metabolismo , Tinción con Nitrato de PlataRESUMEN
Gout is a clinical syndrome in which tissue damage is induced by a chronic metabolic disorder associated with increased concentrations of uric acid in the blood. The study investigated the hypouricemic effects of anthocyanin extracts from purple sweet potato (APSP), and allopurinol, on serum uric acid levels in hyperuricemic mice. It was found that administration of a single oral dose of 100 mg/kg APSP to such animals reduced the serum uric acid concentration to 4.10 ± 0.04 mg/dL, compared with a concentration of 10.25 ± 0.63 mg/dL in the hyperuricemic control group.
Asunto(s)
Antocianinas/farmacología , Hiperuricemia/tratamiento farmacológico , Ipomoea batatas/química , Extractos Vegetales/farmacología , Alopurinol/farmacología , Animales , Modelos Animales de Enfermedad , Hiperuricemia/inducido químicamente , Masculino , Ratones , Ratones Endogámicos ICR , Ácido Oxónico , Ácido Úrico/sangreRESUMEN
Matrix metalloproteinase (MMP)-9 plays a key role in tumor invasion. Inhibitors of MMP-9 were screened from Metasequoia glyptostroboides (Dawn redwood) and one potent inhibitor, isoginkgetin, a biflavonoid, was identified. Noncytotoxic levels of isoginkgetin decreased MMP-9 production profoundly, but up-regulated the level of tissue inhibitor of metalloproteinase (TIMP)-1, an inhibitor of MMP-9, in HT1080 human fibrosarcoma cells. The major mechanism of Ras-dependent MMP-9 production in HT1080 cells was phosphatidylinositol 3-kinase (PI3K)/Akt/nuclear factor-kappaB (NF-kappaB) activation. Expression of dominant-active H-Ras and p85 (a subunit of PI3K) increased MMP-9 activity, whereas dominant-negative forms of these molecules decreased the level of MMP-9. H-Ras did not increase MMP-9 in the presence of a PI3K inhibitor, LY294002, and a NF-kappaB inhibitor, SN50. Further studies showed that isoginkgetin regulated MMP-9 production via PI3K/Akt/NF-kappaB pathway, as evidenced by the findings that isoginkgetin inhibited activities of both Akt and NF-kappaB. PI3K/Akt is a well-known key pathway for cell invasion, and isoginkgetin inhibited HT1080 tumor cell invasion substantially. Isoginkgetin was also quite effective in inhibiting the activities of Akt and MMP-9 in MDA-MB-231 breast carcinomas and B16F10 melanoma. Moreover, isoginkgetin treatment resulted in marked decrease in invasion of these cells. In summary, PI3K/Akt is a major pathway for MMP-9 expression and isoginkgetin markedly decreased MMP-9 expression and invasion through inhibition of this pathway. This suggests that isoginkgetin could be a potential candidate as a therapeutic agent against tumor invasion.
Asunto(s)
Biflavonoides/farmacología , Flavonoides/farmacología , Metaloproteinasa 9 de la Matriz/metabolismo , Neoplasias/enzimología , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de Señal/efectos de los fármacos , Neoplasias de la Mama/metabolismo , Relación Dosis-Respuesta a Droga , Femenino , Fibrosarcoma/metabolismo , Humanos , Inhibidores de la Metaloproteinasa de la Matriz , Invasividad Neoplásica , Inhibidores de las Quinasa Fosfoinosítidos-3 , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/antagonistas & inhibidores , Inhibidor Tisular de Metaloproteinasa-1/metabolismo , Células Tumorales Cultivadas , Quinasa de Factor Nuclear kappa BRESUMEN
Tungtungmadic acid (3-caffeoyl-4-dihydrocaffeoyl quinic acid) is a new chlorogenic acid derivative that was isolated from the Salicornia herbacea. The structure of tungtungmadic acid was determined using chemical and spectral analysis. The antioxidant activity of tungtungmadic acid was evaluated using various antioxidant assays, including free radical scavenging, lipid peroxidation and hydroxyl radical-induced DNA strand breaks assays. Tungtungmadic acid (IC50 = 5.1 microM and 9.3 microM) was found to have higher antioxidant activity in the DPPH scavenging assay as well as in the iron-induced liver microsomal lipid peroxidation system. In addition, the tungtungmadic acid was also effective in protecting the plasmid DNA against strand breakage induced by hydroxyl radicals.
Asunto(s)
Amaranthaceae/química , Antioxidantes/aislamiento & purificación , Ácido Clorogénico/aislamiento & purificación , Animales , Antioxidantes/química , Antioxidantes/farmacología , Ácido Clorogénico/química , Ácido Clorogénico/farmacología , Daño del ADN/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Peroxidación de Lípido/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos ICR , Microsomas Hepáticos/efectos de los fármacos , Microsomas Hepáticos/metabolismo , Estructura Molecular , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/farmacologíaRESUMEN
Excessive nitric oxide production by inducible nitric oxide synthase (iNOS) in stimulated inflammatory cells is thought to be a causative factor of cellular injury in cases of inflammation. In recent studies, it has been shown that kahweol, coffee-specific diterpene, exhibit chemoprotective effects. In this study, we investigated the effects of kahweol on the production of and the expression of inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-activated RAW 264.7 macrophages. The nitrite production induced by LPS was markedly reduced in a dose-dependent manner. In addition, kahweol suppressed the expression of iNOS protein and iNOS mRNA. Since iNOS transcription has been shown to be under the control of the transcription factor, NF-kappaB, the effects of kahweol on NF-kappaB activation were examined. Transient transfection experiments showed that kahweol inhibited NF-kappaB-dependent transcriptional activity. Moreover, electrophoretic mobility shift assay experiments indicated that kahweol blocked the LPS-induced activation of NF-kappaB. The results of these studies suggest that the suppression of the transcriptional activation of iNOS by kahweol might be mediated through the inhibition of NF-kappaB activation. Taken together, the results of our study provide evidence that kahweol possess an anti-inflammatory potential, which constitutes a previously unrecognized biologic activity, and which may provide new insights into the inflammatory process.
Asunto(s)
Diterpenos/farmacología , Macrófagos/enzimología , Óxido Nítrico Sintasa/biosíntesis , Óxido Nítrico Sintasa/efectos de los fármacos , Animales , Relación Dosis-Respuesta a Droga , Inducción Enzimática/efectos de los fármacos , Inflamación , Ratones , Transcripción Genética/efectos de los fármacosRESUMEN
A-factor (2-isocapryloyl-3R-hydroxymethyl-gamma-butyrolactone) acts as a microbial hormone that induces morphological development and secondary metabolism in Streptomyces griseus. A diffusible yellow pigment is produced by S. griseus in an A-factor-dependent manner under phosphate depletion. Detailed analysis of the pigment production by S. griseus cultivated in minimal liquid medium containing different concentrations of phosphate showed that the pigment was actively produced in the presence of low concentrations of phosphate and the production of the pigment was completely repressed in the presence of 2.5 mM KH2PO4. HPLC analysis of the culture supernatant showed that the pigment consisted of two major, structurally related compounds and they were produced at different ratios depending on the concentration of phosphate in the medium. The structures of the two major compounds, designated as grixazone A and B, were determined by spectroscopic analyses as 1-[[2-(acetylamino)-2-carboxyethyl]thio]-2-amino-3-oxo-8-formyl-3H-phenoxiazine and 1-[[2-(acetylamino)-2-carboxyethyl]thio]-2-amino-3-oxo-8-carboxyl-3H-phenoxiazine, respectively. Grixazone A was a novel compound, although grixazone B was reported in a patent as a parasiticide produced by Streptomyces sp. DSM3813.
Asunto(s)
Pigmentos Biológicos/biosíntesis , Streptomyces griseus/metabolismo , Cromatografía Líquida de Alta Presión , Oxazinas/química , Oxazinas/aislamiento & purificación , Fosfatos/deficiencia , Pigmentos Biológicos/química , Pigmentos Biológicos/aislamiento & purificación , Streptomyces griseus/fisiologíaRESUMEN
SCOPE: Increasing evidence indicates that polyphenols may protect against metabolic disease through activating AMP-activated protein kinase (AMPK). The aims of our study were to provide new data on the molecular mechanism(s) underlying the role of the phenolic compound, 3-caffeoyl, 4-dihydrocaffeoylquinic acid (CDCQ) from Salicornia herbacea, in the prevention of high glucose-induced lipogenesis in human HepG2 cells. METHODS AND RESULTS: Nile red staining assays were used to demonstrate lipid accumulation in the cells. Expression of sterol regulatory element-binding protein-1c (SREBP-1c) and fatty acid synthase (FAS) gene at the levels of promoter activity, mRNA, and protein was demonstrated using transient transfection assays, quantitative RT-PCR, and Western blot analyses, respectively. We found that CDCQ suppressed high glucose-induced lipid accumulation in HepG2 cells. CDCQ strongly inhibited high glucose-induced FAS expression by modulating SREBP-1c activation. Moreover, the use of both a specific inhibitor and liver kinase B1 (LKB1)-siRNA transfected HepG2 cells showed that CDCQ activated AMPK via silent information regulator T1 (SIRT1) or LKB1 in HepG2 cells. CONCLUSION: These results indicate that CDCQ prevented lipid accumulation by blocking the expression of SREBP-1c and FAS through LKB1/SIRT1 and AMPK activation in HepG2 cells, suggesting that CDCQ plays a potential role in the prevention of lipogenesis by AMPK activation.
Asunto(s)
Proteínas Quinasas Activadas por AMP/metabolismo , Chenopodiaceae/química , Ácido Clorogénico/análogos & derivados , Glucosa/efectos adversos , Lipogénesis/efectos de los fármacos , Proteínas Serina-Treonina Quinasas/metabolismo , Quinasas de la Proteína-Quinasa Activada por el AMP , Ácido Clorogénico/farmacología , Activación Enzimática/efectos de los fármacos , Acido Graso Sintasa Tipo I/genética , Acido Graso Sintasa Tipo I/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Células Hep G2/efectos de los fármacos , Células Hep G2/metabolismo , Humanos , Metabolismo de los Lípidos/efectos de los fármacos , Regiones Promotoras Genéticas , ARN Interferente Pequeño , Transducción de Señal/efectos de los fármacos , Sirtuina 1/genética , Sirtuina 1/metabolismo , Proteína 1 de Unión a los Elementos Reguladores de Esteroles/genética , Proteína 1 de Unión a los Elementos Reguladores de Esteroles/metabolismoRESUMEN
Chives have been used both as food and as medicine. Previously, two fibrinolytic enzymes, ATFE-I (90 kDa) and ATFE-II (55 kDa), were identified in chives (Allium tuberosum), a perennial herb. In the present work, ATFE-II was purified by ion-exchange chromatography followed by gel filtration. In addition, the enzyme properties of ATFE-I and ATFE-II were compared. The molecular mass and isoelectric point (pI value) of ATFE-II were 55 kDa and pI 4.0, respectively, as revealed using one- or two-dimensional fibrin zymography. ATFE-II was optimally active at pH 7.0 and 45°C. ATFE-II degraded the Aα-chain of human fibrinogen but did not hydrolyze the Bß-chain or the γ-chain, indicating that the enzyme is an α-fibrinogenase. The proteolytic activity of ATFE-II was completely inhibited by 1 mM leupeptin, indicating that the enzyme belongs to the cysteine protease class. ATFE-II was also inhibited by 1 mM Fe²(+). ATFE-II exhibited high specificity for MeO-Suc-Arg-Pro-Tyr-p-nitroaniline (S-2586), a synthetic chromogenic substrate of chymotrypsin. Thus proteolytic enzymes from A. tuberosum may be useful as thrombolytic agents.
Asunto(s)
Cebollino/enzimología , Cisteína Endopeptidasas , Descubrimiento de Drogas , Fibrinólisis , Fibrinolíticos , Componentes Aéreos de las Plantas/enzimología , Proteínas de Plantas , Cisteína Endopeptidasas/química , Cisteína Endopeptidasas/aislamiento & purificación , Cisteína Endopeptidasas/metabolismo , Inhibidores de Cisteína Proteinasa/farmacología , Compuestos Ferrosos/farmacología , Fibrinógeno/metabolismo , Fibrinolíticos/química , Fibrinolíticos/aislamiento & purificación , Fibrinolíticos/metabolismo , Concentración de Iones de Hidrógeno , Hidrólisis , Punto Isoeléctrico , Isoenzimas/antagonistas & inhibidores , Isoenzimas/química , Isoenzimas/aislamiento & purificación , Isoenzimas/metabolismo , Leupeptinas/farmacología , Peso Molecular , Oligopéptidos/metabolismo , Proteínas de Plantas/antagonistas & inhibidores , Proteínas de Plantas/química , Proteínas de Plantas/aislamiento & purificación , Proteínas de Plantas/metabolismo , Especificidad por Sustrato , Temperatura , Terapia Trombolítica , Trombosis/tratamiento farmacológicoRESUMEN
Salicornia herbacea (S. herbacea), an annual herb that grows in the salt marshes of the Korean peninsula, has been used as a folk medicine to treat a variety of diseases such as constipation, obesity, diabetes, and cancer. However, the effect of S. herbacea on inflammation is unclear. In the present study, we investigated the effects of a novel chlorogenic acid, 3-caffeoyl-4-dicaffeoylquinic acid (CDCQ), isolated from S. herbacea, on cyclooxygenase-2 (COX-2) expression in murine macrophage RAW 264.7 cells. Phorbol 12-myristate 13-acetate (PMA) induces COX-2 expression and production of prostaglandin E(2) (PGE(2)). PMA-induced COX-2 protein, gene expression and PGE(2) production were significantly inhibited by CDCQ in a dose-dependent manner. Transfection of hCOX-2, as well as of deletion and mutation promoter constructs, revealed that the CCAAT/enhancer-binding protein (C/EBP) and activator protein-1 (AP-1) predominantly contributed to the effects of CDCQ. In addition, electrophoretic mobility shift assays and transfection results showed that CDCQ directly inhibited PMA-induced C/EBP and AP-1 transcription and binding activity. CDCQ also remarkably reduced PMA-induced C/EBPbeta and c-jun protein expression. Furthermore, CDCQ significantly inhibited PMA-induced activation of the mitogen-activated protein kinases (MAP kinases), JNK and p38. These findings demonstrate that CDCQ effectively attenuates COX-2 production, and enhance our understanding of the anti-inflammatory properties of CDCQ.
Asunto(s)
Chenopodiaceae/química , Ácido Clorogénico/toxicidad , Ciclooxigenasa 2/metabolismo , Macrófagos/enzimología , Ésteres del Forbol/farmacología , Animales , Proteína beta Potenciadora de Unión a CCAAT/metabolismo , Línea Celular Tumoral , Ácido Clorogénico/química , Ácido Clorogénico/aislamiento & purificación , Ciclooxigenasa 2/genética , Dinoprostona/metabolismo , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Ratones , Proteínas Proto-Oncogénicas c-jun/metabolismo , Factor de Transcripción AP-1/metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
In this study, we determined the effects of a novel chlorogenic acid, 3-caffeoyl, 4-dicaffeoylquinic acid (CDCQ) isolated from Salicornia herbacea, on tumor invasion and migration in human fibrosarcoma HT-1080 cells and investigated the possible mechanism(s) involved. CDCQ reduced the phorbol myristate acetate (PMA)-induced activation of matrix metalloproteinase (MMP)-9 and MMP-2 and inhibited cell invasion and migration. CDCQ suppressed PMA-induced expression of MMP-9 mRNA and protein by suppressing the transcription factor AP-1, without changing the level of tissue inhibitor of metalloproteinase (TIMP)-1. CDCQ-inhibited PMA-induced MMP-2 expression by suppressing membrane-type 1 MMP (MT1-MMP), but did not alter the TIMP-2 level. CDCQ also inhibited the PMA-induced nuclear translocation of c-Jun and c-Fos, which are upstream of PMA-induced MMP-9 expression. Furthermore, CDCQ strongly repressed PMA-induced phosphorylation of ERK, p38 MAPK, and JNK, which are dependent on the PKCdelta pathway. In conclusion, we demonstrated that the anti-invasive effects of CDCQ occur through the inhibition of AP-1 and signaling pathways involving PKCdelta and three MAPKs, leading to the downregulation of MMP-9 expression. Thus, CDCQ is an effective anti-metastatic agent that functions by downregulating MMP-9 gene expression.
Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Chenopodiaceae/química , Ácido Clorogénico/análogos & derivados , Expresión Génica/efectos de los fármacos , Metaloproteinasa 9 de la Matriz/genética , Proteína Quinasa C-delta/metabolismo , Antineoplásicos Fitogénicos/aislamiento & purificación , Western Blotting , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Ácido Clorogénico/aislamiento & purificación , Ácido Clorogénico/farmacología , Humanos , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Salicornia herbacea has been used as a folk medicine for disorders such as constipation, obesity, diabetes, and cancer. Recent studies have shown that S. herbacea has antioxidative, anti-inflammatory, immunomodulatory, antihyperglycemic, and antihyperlipidemic activities. In the present work, we investigated the protective effects of the chlorogenic acid derivative, 3-caffeoyl, 4-dihydrocaffeoyl quinic acid (CDCQ), which was isolated from S. herbacea, against tert-butyl hydroperoxide (t-BHP)-induced hepatotoxicity in Hepa1c1c7 cells. Pretreatment of Hepa1c1c7 cells with CDCQ significantly reduced t-BHP-induced generation of ROS, caspase-3 activation, and subsequent cell death. Also, CDCQ up-regulated heme oxygenase-1 (HO-1) expression, which conferred cytoprotection against oxidative injury induced by t-BHP. Moreover, CDCQ-induced nuclear translocation of the transcription factor NF-E2-related factor 2 (Nrf2), which is upstream of CDCQ-induced HO-1 expression, and PI3K/Akt activation, a pathway that is involved in induced Nrf2 nuclear translocation. Taken together, these results suggest that the protective effects of CDCQ against t-BHP-induced hepatotoxicity may be due, at least in part, to its ability to scavenge ROS and to regulate the antioxidant enzyme HO-1 via the PI3K/Akt-Nrf2 signaling pathways.
Asunto(s)
Antioxidantes/farmacología , Chenopodiaceae/química , Ácido Clorogénico/análogos & derivados , Estrés Oxidativo , terc-Butilhidroperóxido/toxicidad , Western Blotting , Caspasa 3/metabolismo , Línea Celular , Ácido Clorogénico/farmacología , Humanos , Inmunohistoquímica , Etiquetado Corte-Fin in Situ , Factor 2 Relacionado con NF-E2/genética , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , ARN Interferente Pequeño , Especies Reactivas de Oxígeno/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Because the activation of matrix metalloproteinases (MMP) is a key factor in the metastatic process, compounds with the ability to inhibit MMP activity have a potential in the treatment of tumor. From the examination of 2000 plant extracts, obovatal isolated from the extract of the leaves of Magnolia obovata THUNB was a potent inhibitor of MMP-2 enzyme in vitro. In human fibrosarcoma cells (HT1080) activated with MMP-2, obovatal inhibited MMP-2 enzyme activity and expression. In addition, the compound blocked migration and invasion of the cells. This study demonstrates that obovatal exerts its anticancer effects through blocking migration and invasion by inhibition of MMP-2 expression and activity, and also will be a good lead molecule for the development of anti-tumor drug.
Asunto(s)
Movimiento Celular/efectos de los fármacos , Inhibidores de la Metaloproteinasa de la Matriz , Inhibidores de Proteasas/farmacología , Línea Celular Tumoral , Humanos , Magnolia/química , Invasividad Neoplásica , Extractos Vegetales/farmacología , Hojas de la Planta/química , Inhibidores de Proteasas/aislamiento & purificaciónRESUMEN
Two new eucosterol oligoglycosides, 15-deoxo-30-hydroxyeucosterol 3-O-alpha-L-rhamnopyranosyl-(1-->2)-[(beta-D-glucopyranosyl-(1-->3)]-beta-D-glucopyranosyl-(1-->2)-alpha-L-arabinopyranosyl-(1-->6)-beta-D-glucopyranoside (scillanoside L-1, 1) and 3beta,31-dihydroxy-17alpha,23-epoxy-5alpha-lanost-8-en-23,26-olactone 3-O-alpha-L-rhamnopyranosyl-(1-->2)-[beta-D-glucopyranosyl-(1-->3)]-beta-D-glucopyranosyl-(1-->2)-alpha-L-arabinopyranosyl-(1-->6)-beta-D-glucopyranoside (scillanoside L-2, 2), were isolated from the bulbs of Scilla scilloides, together with four that were known (3-6), have been isolated from the bulbs of Scilla scilloides. The structures of the new compounds were determined on the basis of spectroscopic and chromatographic methods, and some chemical transformations were discussed. Amongst the isolated compounds, 3 showed the most significant cytotoxicity against tumor cells tested several types with ED(50) value of 1.53-3.06 nM. In vivo experiments, 3 apparently increased the life span of mice bearing Sarcoma 180 tumor cell with T/C value of 239% at dose of 3 mg/kg.
Asunto(s)
Antineoplásicos Fitogénicos/química , Antineoplásicos Fitogénicos/farmacología , Oligosacáridos/química , Oligosacáridos/aislamiento & purificación , Fitosteroles/química , Fitosteroles/aislamiento & purificación , Scilla/química , Animales , Antineoplásicos Fitogénicos/aislamiento & purificación , Cromatografía en Capa Delgada , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Hidrólisis , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Melanoma Experimental/tratamiento farmacológico , Melanoma Experimental/patología , Modelos Moleculares , Conformación Molecular , Oligosacáridos/farmacología , Fitosteroles/farmacología , Sarcoma 180/tratamiento farmacológico , Sarcoma 180/patología , Espectrofotometría Infrarroja , Células Tumorales CultivadasRESUMEN
The protective effects of 18beta-glycyrrhetinic acid (GA), the aglycone of glycyrrhizin (GL) derived from licorice, on carbon tetrachloride-induced hepatotoxicity and the possible mechanisms involved in this protection were investigated in mice. Pretreatment with GA prior to the administration of carbon tetrachloride significantly prevented an increase in serum alanine, aspartate aminotransferase activity and hepatic lipid peroxidation in a dose-dependent manner. In addition, pretreatment with GA also significantly prevented the depletion of glutathione (GSH) content in the livers of carbon tetrachloride-intoxicated mice. However, reduced hepatic GSH levels and glutathione-S-transferase activities were unaffected by treatment with GA alone. Carbon tetrachloride-induced hepatotoxicity was also prevented, as indicated by a liver histopathologic study. The effects of GA on the cytochrome P450 (P450) 2E1, the major isozyme involved in carbon tetrachloride bioactivation, were also investigated. Treatment of mice with GA resulted in a significant decrease of the P450 2E1-dependent hydroxylation of p-nitrophenol and aniline in a dose-dependent manner. Consistent with these observations, the P450 2E1 expressions were also decreased, as determined by immunoblot analysis. GA also showed antioxidant effects upon FeCl(2)-ascorbate-induced lipid peroxidation in mice liver homogenate and upon superoxide radical scavenging activity. These results show that protective effects of GA against the carbon tetrachloride-induced hepatotoxicity may be due to its ability to block the bioactivation of carbon tetrachloride, primarily by inhibiting the expression and activity of P450 2E1, and its free radical scavenging effects.