RESUMEN

Chloroquine (CQ) is used to treat malaria and a variety of inflammatory diseases including systemic lupus erythematosus and rheumatoid arthritis. However, CQ is known to cause cytotoxicity of which mechanism is still uncertain. This study investigated the molecular mechanism responsible for the cell death in CQ-treated A172 human glioblastoma cells. CQ-induced apoptotic cell death of the cells in a time- and concentration-dependent manner. CQ also increased the production of nitric oxide in the cells. However, the pretreatment with aminoguanidine (AG) and N-Omega-nitro-l-arginine methyl ester (NAME), nitric oxide synthase inhibitors, did not block the CQ-induced cell death. In contrast to NO level increase, the level of intracellular reactive oxygen species (ROS) and their extracellular release were transiently and mildly increased by CQ. In addition, CQ depleted cellular GSH content, which was accompanied with time-dependent increase in GSH peroxidase without any significant change in GSH reductase activity. Glutathione (GSH) S-transferase activity was only transiently increased at 15 min treatment with CQ. Furthermore, the CQ-induced cell death was significantly suppressed when intracellular GSH decrease was prevented by the pretreatment with N-acetylcysteine (NAC) or glutathione ethylester (GSH-EE). At the same time, the pretreatment of the cells with NAC and GSH-EE significantly blocked the CQ-induced NO increase, representing that CQ-induced NO increase was resulted from the depletion of GSH. CQ also induced time-dependent increase in Bax level and caspase-3 activity with no change in Bcl-2 level. Overall, these results suggest that CQ-induced NO increase and cell death are dependent on GSH depletion, the cellular redox changes.

Asunto(s)

Antimaláricos/toxicidad , Antirreumáticos/toxicidad , Apoptosis , Cloroquina/toxicidad , Glutatión/metabolismo , Óxido Nítrico/metabolismo , Caspasa 3/metabolismo , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Glioblastoma , Glutatión Peroxidasa/metabolismo , Glutatión Reductasa/metabolismo , Glutatión Transferasa/metabolismo , Humanos , Nitritos/metabolismo , Oxidación-Reducción , Especies Reactivas de Oxígeno/metabolismo , Proteína X Asociada a bcl-2/metabolismo

RESUMEN

Asiatic acid, a pentacyclic triterpene, has been reported to induce apoptosis of various human cancer cells. In the present study, we assessed the anti-tumor promoting effect of asiatic acid against 12-O-tetradecanoylphorbol 13-acetate (TPA)-mediated skin tumorigenesis in 7,12-dimethylbenz[a]anthracene (DMBA)-initiated ICR mice. Topical application of asiatic acid prior to each application of TPA resulted in a significant reduction in skin tumor formation. We also found that pre-application of asiatic acid alleviated TPA-induced [3H]thymidine incorporation, which is a conventional marker for skin tumor promotion. In addition, asiatic acid inhibited the TPA-induced generation of nitric oxide (NO) and expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2), which are known to play important roles in tumor growth, especially in the promotion stage. In addition, topical application of aminoguanidine (AG), a selective iNOS inhibitor, and N(G)-nitro-L-arginine-methyl ester (NAME), another iNOS inhibitor, 30 min prior to TPA treatment significantly inhibited the TPA-induced COX-2 expression. These results suggest that asiatic acid may exert anti-tumorigenesis through inhibitory actions in NO and COX-2 signals.

Asunto(s)

Anticarcinógenos/farmacología , Papiloma/prevención & control , Neoplasias Cutáneas/prevención & control , Piel/efectos de los fármacos , Triterpenos/farmacología , 9,10-Dimetil-1,2-benzantraceno , Animales , Anticarcinógenos/uso terapéutico , Carcinógenos , Transformación Celular Neoplásica/efectos de los fármacos , Cocarcinogénesis , Ciclooxigenasa 2/biosíntesis , Relación Dosis-Respuesta a Droga , Inducción Enzimática/efectos de los fármacos , Femenino , Ratones , Ratones Endogámicos ICR , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/biosíntesis , Papiloma/inducido químicamente , Triterpenos Pentacíclicos , Piel/metabolismo , Neoplasias Cutáneas/inducido químicamente , Acetato de Tetradecanoilforbol , Factores de Tiempo , Triterpenos/uso terapéutico

RESUMEN

To develop a novel clotrimazole-loaded poloxamer-based suppository with enhanced anti-tumor activity and alleviated hepatotoxicity, the melting point of various formulations composed of P 188 and propylene glycol were investigated. The dissolution and anti-tumor activity of clotrimazole delivered by the poloxamer-based suppository was performed. Furthermore, the hepatotoxicity of clotrimazole was carried out after its rectal administration compared to oral administration in mice. The poloxamer mixtures composed of P 188 and propylene glycol were homogeneous phases. P 188 greatly affected the melting point of poloxamer mixtures. In particular, the poloxamer mixture [P 188/propylene glycol (70%/30%)] with the melting point of about 32 degrees C was a solid form at room temperature and instantly melted at physiological temperature. The ratio of P 188/propylene glycol greatly affected the dissolution rates of clotrimazole from poloxamer-based suppository. Dissolution mechanism analysis showed the dissolution rate of clotrimazole from poloxamer-based suppositories was independent of the time. The clotrimazole-loaded suppository with P 188 and propylene glycol could not irritate or damage the rectal tissues of rats and gave the improved anti-tumor activity in a dose-dependent manner at mouse. Furthermore, its rectal administration decreased the hepatotoxicity compared to oral administration. Thus, the poloxamer-based solid suppository system with clotrimazole/P 188/propylene glycol was an effective rectal dosage form for the treatment of tumors with alleviated adverse effects.

Asunto(s)

Antineoplásicos/administración & dosificación , Clotrimazol/administración & dosificación , Hígado/efectos de los fármacos , Animales , Clotrimazol/toxicidad , Geles , Masculino , Ratones , Ratones Endogámicos BALB C , Poloxámero/administración & dosificación , Propilenglicol/administración & dosificación , Ratas , Ratas Sprague-Dawley , Solubilidad , Supositorios

RESUMEN

The tumoricidal and apoptosis-inducing activities of 5-fluorouracil (5-FU) have been demonstrated in experimental and clinical investigations. Clinically, the 5-FU suppository form has been widely adopted for its advantages of less systemic toxicity, higher local tissue concentrations, and reduced first-pass effect. In this study, we investigated the feasibility of rectal administration of 5-FU suppository based on poloxamer 188 (P188) and propylene glycol (PG) and its anticancer effect on the murine experimental cancer models. The rectal suppository was made with 70% P188 and 30% PG, which was a solid phase at room temperature and instantly melted at physiological temperature. The treatment with the 5-FU suppository was more effective than the oral route in decreasing the volume of rectal cancer in mice. In addition, the survival rate of the mice with rectal cancer was higher in the group treated with the 5-FU suppository than in the group treated with 5-FU orally. Furthermore, in mice skin cancers induced by inoculation of murine CT-26 colon carcinoma cells, the anticancer effect of 5-FU was significantly enhanced by the rectal administration of the suppository than by oral treatment. Taken together, the results suggest that a poloxamer gel system with 5-FU/P188/PG is an effective rectal dosage form for the treatment of both rectal and non-rectal cancers.

Asunto(s)

Antimetabolitos Antineoplásicos/administración & dosificación , Antimetabolitos Antineoplásicos/farmacología , Fluorouracilo/administración & dosificación , Fluorouracilo/farmacología , Poloxámero , Propilenglicol , Administración Rectal , Alanina Transaminasa/sangre , Animales , Aspartato Aminotransferasas/sangre , Línea Celular Tumoral , Peroxidación de Lípido/efectos de los fármacos , Ratones , Ratones Endogámicos BALB C , Trasplante de Neoplasias , Vehículos Farmacéuticos , Neoplasias del Recto/tratamiento farmacológico , Neoplasias Cutáneas/tratamiento farmacológico , Solubilidad , Supositorios

RESUMEN

Nonsteroidal anti-inflammatory drugs (NSAIDs) are the most frequently prescribed drug for the treatment of inflammation and pain. However, conventional NSAIDs and selective COX-2 inhibitors have shown many side effects such as gastric mucosal damage and cardiovascular problems. Recently, the use of dual acting inhibitors of cyclooxygenases (COX) and lipoxygenase (LOX) has been highlighted for their minimized side effects compared to NSAIDs. The objective of the present study was to examine the efficacy and the gastric side effects of 1-furan-2-yl-3-pyridin-2-yl-propenone (FPP-3), a synthetic dual inhibitor of COX/5-LOX. Indomethacin (1-50 mg/kg, p.o.), a non-selective COX inhibitor, and FPP-3 (0.5-50 mg/kg, p.o.), a dual inhibitor, significantly suppressed the carrageen-induced paw edema with different pharmacological profiles. The concentrations of FPP-3 and indomethacin showing 50% inhibition of the maximum paw edema in rats were 10 mg/kg and 20 mg/kg, respectively. More importantly, there were no gastric ulcers formed in FPP-3-treated rats and mice, whereas indomethacin caused gastric mucosal bleeding in a concentration-dependent manner. In addition, FPP-3 showed an analgesic effect in acetic acid-induced writhing response in mice in a dose-dependent manner. The results suggest that FPP-3 may have a benefit in combatting inflammation and pain by dual inhibition of COX and LOX.

Asunto(s)

Antiinflamatorios no Esteroideos/farmacología , Inhibidores de la Ciclooxigenasa/farmacología , Furanos/farmacología , Mucosa Gástrica/efectos de los fármacos , Propano/análogos & derivados , Piridinas/farmacología , Úlcera Gástrica/inducido químicamente , Animales , Antiinflamatorios no Esteroideos/efectos adversos , Antiinflamatorios no Esteroideos/uso terapéutico , Inhibidores de la Ciclooxigenasa/efectos adversos , Inhibidores de la Ciclooxigenasa/uso terapéutico , Modelos Animales de Enfermedad , Edema/tratamiento farmacológico , Furanos/efectos adversos , Furanos/uso terapéutico , Mucosa Gástrica/patología , Ratones , Ratones Endogámicos ICR , Propano/efectos adversos , Propano/farmacología , Propano/uso terapéutico , Piridinas/efectos adversos , Piridinas/uso terapéutico , Ratas , Ratas Sprague-Dawley , Úlcera Gástrica/patología

RESUMEN

Although alpha-chaconine, one of the two major potato trisaccharide glycoalkaloids, have shown cytotoxic effects on human cancer cells, the exact mechanism of this action of alpha-chaconine is not completely understood. In this study, we found that alpha-chaconine induced apoptosis of HT-29 cells in a time- and concentration-dependent manner by using flow cytometric analysis. We also found that caspase-3 activity and the active form of caspase-3 were increased 12 h after alpha-chaconine treatment. Caspase inhibitors, N-Ac-DEVD-CHO and Z-VAD-fmk, prevented alpha-chaconine-induced apoptosis, whereas alpha-chaconine-induced apoptosis was potentiated by PD98059, an extracellular signal-regulated kinase (ERK) inhibitor. However, pretreatment of the cells with LY294002 and SB203580, inhibitors of PI3K and p38, respectively, BAPTA-AM, an intracellular Ca(2+) chelator, and antioxidants such as N-acetylcysteine (NAC) and Trolox had no effect on the alpha-chaconine-induced cell death. In addition, phosphorylation of ERK was reduced by the treatment with alpha-chaconine. Moreover, alpha-chaconine-induced caspase-3 activity was further increased by the pretreatment with PD98059. Thus, the results indicate that alpha-chaconine induces apoptosis of HT-29 cells through inhibition of ERK and, in turn, activation of caspase-3.

Asunto(s)

Apoptosis/efectos de los fármacos , Caspasas/metabolismo , Inhibidores Enzimáticos/farmacología , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , Solanina/análogos & derivados , Caspasa 3 , Activación Enzimática/efectos de los fármacos , Células HT29 , Humanos , Proteína Quinasa 1 Activada por Mitógenos/antagonistas & inhibidores , Proteína Quinasa 3 Activada por Mitógenos/antagonistas & inhibidores , Fosforilación/efectos de los fármacos , Solanina/farmacología

RESUMEN

The volatile extract from dried pericarp of Zanthoxylum schinifolium that was obtained by simultaneous distillation with dichloromethane and water was composed of 29.9% geranyl acetate, 15.8% citronella, 15.4% sabinene and the minor volatile components included beta-myrcene, linalool, (-)-isopulegol, citronellyl acetate, 1,4-dimethyl pyrazole, alpha-terpinene, 3-methyl-6-(1-methylethyl)-2-cyclo-hexene-1-o1 and trans-geraniol. The volatile extract decreased the cell viability and induced apoptotic death in HepG2 human hepatoma cells in a concentration- and time-related manner. In addition, the volatile extract increased the production of reactive oxygen species in a dose-dependent manner. Pretreatment of the cells with Trolox, a well-known antioxidant, significantly suppressed the generation of reactive oxygen species and cell death induced by the extract. However, caspase-3 activity was not changed in the extract-treated cells, suggesting that the extract-induced apoptosis of HepG2 cells is caspase-3 independent. Furthermore, in nude mice inoculated with Huh-7 human hepatoma cells, the extract significantly inhibited tumor development. These results suggest that the volatile extract from Zanthoxylum schinifolium pericarpium is a good candidate for hepatocellular carcinoma (HCC) therapy and that reactive oxygen species are the key signaling molecules in the volatile extract-induced cell death in HepG2 cells.

Asunto(s)

Apoptosis/efectos de los fármacos , Carcinoma Hepatocelular/metabolismo , Aceites Volátiles/farmacología , Especies Reactivas de Oxígeno/metabolismo , Zanthoxylum , Animales , Apoptosis/fisiología , Carcinoma Hepatocelular/tratamiento farmacológico , Relación Dosis-Respuesta a Droga , Femenino , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Aceites Volátiles/aislamiento & purificación , Aceites Volátiles/uso terapéutico , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/farmacología , Extractos Vegetales/uso terapéutico , Ensayos Antitumor por Modelo de Xenoinjerto

RESUMEN

Matrix metalloproteinases (MMPs), a key component in photoaging of the skin due to exposure to ultraviolet A, appear to be increased by ultraviolet A irradiation-associated generation of reactive oxygen species. In this study, we investigated the effects of synthetic rutaecarpine, which is also found in Evodia rutaecarpa, on the ultraviolet A-induced changes in the expression of gelatinases: matrix metalloproteinase (MMP)-2 and MMP-9 using HaCaT human keratinocytes as a model cellular system. Ultraviolet A irradiation of HaCaT cells increased the gelatinolytic activities of MMP-2 and MMP-9, which was significantly suppressed by the pretreatment with rutaecarpine. In addition, rutaecarpine significantly suppressed the ultraviolet A-induced enhanced expression of MMP-2 and MMP-9 proteins and mRNAs. Rutaecarpine also inhibited the H2O2-induced increase in the expression of MMP-2 and MMP-9. Furthermore, rutaecarpine decreased the ultraviolet A-induced increased generation of reactive oxygen species. Taken together, these results suggest that rutaecarpine inhibited ultraviolet A-induced reactive oxygen species generation, resulting in the enhanced expression of MMP-2 and MMP-9 in human skin cells. These results further suggest that ruetaecarpine may be useful in the prevention of ultraviolet A-induced photoaging.

Asunto(s)

Alcaloides/farmacología , Queratinocitos/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Acetilcisteína/farmacología , Western Blotting , Línea Celular , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/efectos de la radiación , Activación Enzimática/efectos de los fármacos , Activación Enzimática/efectos de la radiación , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Regulación Enzimológica de la Expresión Génica/efectos de la radiación , Humanos , Peróxido de Hidrógeno/farmacología , Alcaloides Indólicos , Queratinocitos/metabolismo , Queratinocitos/efectos de la radiación , Metaloproteinasa 2 de la Matriz/genética , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/genética , Metaloproteinasa 9 de la Matriz/metabolismo , Quinazolinas , ARN Mensajero/genética , ARN Mensajero/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transducción de Señal/efectos de la radiación , Tretinoina/farmacología , Rayos Ultravioleta