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BACKGROUND/AIM: The COVID-19 pandemic led to the rapid spread of the use of ultraviolet C (UVC) sterilizers in many public facilities. Considering the harmful effects of prolonged exposure to UVC, manufacturing of safe skin care products is an important countermeasure. In continuation of our recent study of water-soluble herbal extracts, the present study aimed at searching for anti-UVC components from fat-soluble herbal extracts. MATERIALS AND METHODS: Human dermal fibroblast and melanoma cells were exposed to UVC (1.193 W/m2) for 3 min. Viable cell number was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Cell-cycle analysis was performed using a cell sorter. UVC-protective activity was quantified by the selective index (SI), i.e., the ratio of the 50% cytotoxic concentration for unirradiated cells to the concentration that restored viability of UVC-treated cells by 50%. RESULTS: Only lemongrass extract, among 12 fat-soluble herbal extracts, showed significant anti-UVC activity, comparable to that of lignified materials and tannins, but exceeding that of N-acetyl-L-cysteine and resveratrol. Lemongrass extract was highly cytotoxic, producing a subG1 cell population. During prolonged incubation in culture medium, the anti-UVC activity of lemongrass extract, sodium ascorbate and vanillic acid declined with an approximate half-life of <0.7, 5.4-21.6, and 27.8-87.0 h, respectively. CONCLUSION: Removal of cytotoxic principle(s) from lemongrass extract is crucial to producing long-lasting UVC-protective effects.
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Cymbopogon , Extractos Vegetales , Humanos , Extractos Vegetales/farmacología , Pandemias , Piel , Rayos Ultravioleta/efectos adversosRESUMEN
BACKGROUND/AIM: COVID-19 pandemic caused the rapid dissemination of ultraviolet C (UVC) sterilization apparatuses. Prolonged exposure to UVC, however, may exert harmful effects on the human body. The aim of the present study was to comprehensively investigate the anti-UVC activity of a total of 108 hot-water soluble herb extracts, using human dermal fibroblast and melanoma cell lines, for the future development of skin care products. MATERIALS AND METHODS: Exposure time to UVC was set to 3 min, and cell viability was determined using the MTT assay. Anti-UVC activity was determined using the selective index (SI), a ratio of 50% cytotoxic concentration for unirradiated cells to 50% effective concentration that restored half of the UVC-induced decrease of viability. RESULTS: Dermal fibroblasts at any population doubling level were more resistant to UVC irradiation than melanoma cells. Both 49 herb extracts recommended by Japan Medical Herb Association (JAMHA) and 59 additional herb extracts showed comparable anti-UVC activity. SI values of selected herbs (Butterbur, Cloves, Curry Tree, Evening Primrose, Rooibos, Stevia, Willow) were several-fold lower than those of vitamin C and vanillin. Their potent anti-UVC activity was maintained for at least 6 h post irradiation, but declined thereafter to the basal level, possibly due to cytotoxic ingredients. CONCLUSION: UVC sensitivity may be related to the growth potential of target cells. Removal of cytotoxic ingredients of herb extracts may further potentiate and prolong their anti-UVC activity.
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COVID-19 , Melanoma , Humanos , Pandemias , Línea Celular , Piel , Rayos Ultravioleta/efectos adversos , Melanoma/tratamiento farmacológico , Extractos Vegetales/farmacologíaRESUMEN
The cytotoxicity of a trivalent arsenic derivative (arsenite, AsIII) combined with arenobufagin or gamabufotalin was evaluated in human U-87 glioblastoma cells. Synergistic cytotoxicity with upregulated intracellular arsenic levels was observed, when treated with AsIII combined with arenobufagin instead of gamabufotalin. Apoptosis and the activation of caspase-9/-8/-3 were induced by AsIII and further strengthened by arenobufagin. The magnitude of increase in the activities of caspase-9/-3 was much greater than that of caspase-8, suggesting that the intrinsic pathway played a much more important role in the apoptosis. An increase in the number of necrotic cells, enhanced LDH leakage, and intensified G2/M phase arrest were observed. A remarkable increase in the expression level of γH2AX, a DNA damage marker, was induced by AsIII+arenobufagin. Concomitantly, the activation of autophagy was observed, suggesting that autophagic cell death associated with DNA damage was partially attributed to the cytotoxicity of AsIII+arenobufagin. Suppression of Notch signaling was confirmed in the combined regimen-treated cells, suggesting that inactivation of Jagged1/Notch signaling would probably contribute to the synergistic cytotoxic effect of AsIII+arenobufagin. Given that both AsIII and arenobufagin are capable of penetrating into the blood-brain barrier, our findings may provide fundamental insight into the clinical application of the combined regimen for glioblastoma.
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Antineoplásicos , Arsénico , Arsenitos , Bufanólidos , Glioblastoma , Antineoplásicos/farmacología , Apoptosis , Arsénico/metabolismo , Arsenitos/farmacología , Bufanólidos/farmacología , Caspasa 8/metabolismo , Caspasa 9/metabolismo , Línea Celular , Línea Celular Tumoral , Glioblastoma/tratamiento farmacológico , Glioblastoma/metabolismo , HumanosRESUMEN
An efficient synthetic method for novel 4,4-disubstituted 3,4-dihydropyrimidin-2(1H)-ones 5 and -thiones 6 was developed. The cyclocondensation reaction of O-methylisourea hemisulfate salt 11 with 8 gives a tautomeric mixture of dihydropyrimidines 12 and 13 following acidic hydrolysis of the cyclized products to produce 5 in high yields. Thionation reaction of 5 at the 2-position smoothly proceeds to give 2-thioxo derivatives 6. These compounds 5 and 6, corresponding to the products of a Biginelli-type reaction using urea or thiourea, a ketone and a 1,3-dicarbonyl compound, have long been inaccessible and hitherto unavailable for medicinal chemistry. These methods are invaluable for the synthesis of 5 and 6, which have been inaccessible by conventional methods. Therefore, the synthetic methods established in this study will expand the molecular diversity of their related derivatives. These compounds were also assessed for their antiproliferative effect on a human promyelocytic leukemia cell line, HL-60. Treatment of 10 µM 6b and 6d showed high inhibitory activity similarly to 1 µM all-trans retinoic acid (ATRA), indicating that the 2-thioxo group and length of two alkyl substituents at the 4-position are strongly related to activity.
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Antineoplásicos/farmacología , Cetonas/farmacología , Pirimidinonas/farmacología , Tionas/farmacología , Antineoplásicos/síntesis química , Antineoplásicos/química , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Ensayos de Selección de Medicamentos Antitumorales , Células HL-60 , Humanos , Cetonas/química , Estructura Molecular , Pirimidinonas/síntesis química , Pirimidinonas/química , Relación Estructura-Actividad , Tionas/síntesis química , Tionas/químicaRESUMEN
αglucosidase is a key enzyme that plays a role in glucose absorption in the gastrointestinal tract, and the inhibition of its activity induces the prevention of postprandial hyperglycemia. Several αglucosidase inhibitors have been used as medicines for type 2 diabetes, but a similar effect is observed in natural resources, including traditional herbs and their phytochemicals. To identify the presence of the αglucosidase inhibitory activity in herbs, in which various functional effects have been known to occur, the present study investigated the effects of hotwater extracts of 26 types of herbs on αglucosidase activity in an in vitro assay. The results indicated significant increases in the inhibition of αglucosidase activity in 1,000 µg/ml olive (P<0.01), white willow (P<0.01) and red rooibos hotwater extracts. Furthermore, ≥50% inhibition of αglucosidase activity was determined to be significant in 1,000 µg/ml coltsfoot, green tea and bearberry hotwater extracts. In addition, the effects of bearberry, green tea and coltsfoot hotwater extracts on αglucosidase activity in vivo were evaluated according to the blood glucose levels (BGLs) in maltose and glucose load model rats. It was indicated that the administration of these three herb extracts significantly reduced the increasing BGLs after maltose loading until 0.5 h compared with the control group. However, only coltsfoot extract significantly reduced the increasing BGLs after glucose loading until 0.5 h compared with the control group. Thus, the present results may facilitate the understanding of a novel functionality in traditional herbs, which could be useful for the prevention of disease onset and progression, such as in hyperglycemia and type 2 diabetes.
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Diabetes Mellitus Tipo 2/tratamiento farmacológico , Inhibidores de Glicósido Hidrolasas/administración & dosificación , Plantas Medicinales/química , Agua/administración & dosificación , alfa-Glucosidasas/metabolismo , Animales , Arctostaphylos/química , Aspalathus/química , Diabetes Mellitus Tipo 2/inducido químicamente , Diabetes Mellitus Tipo 2/enzimología , Modelos Animales de Enfermedad , Glucosa/efectos adversos , Inhibidores de Glicósido Hidrolasas/química , Inhibidores de Glicósido Hidrolasas/farmacología , Calor , Masculino , Maltosa/efectos adversos , Olea/química , Extractos Vegetales/administración & dosificación , Extractos Vegetales/química , Extractos Vegetales/farmacología , Ratas , Salix/química , Té/química , Tussilago/química , Agua/química , Agua/farmacologíaRESUMEN
BACKGROUND: Most previous mastic investigators have not considered its potent cytotoxicity that may significantly affect the interpretation of obtained data. In the present study, we re-evaluated several biological activities of mastic extracts, based on chemotherapeutic indexes. MATERIALS AND METHODS: Pulverized mastic gum was extracted with n-hexane and then with ethyl acetate or independently with methanol or n-butanol. Tumor specificity (TS) of the extracts was determined by their cytotoxicity against human malignant and non-malignant cells. Antibacterial activity was determined by their cytotoxicity against bacteria and normal oral cells. Antiviral activity was determined by their protection of viral infection and cytotoxic activity. Cytochrome P-450 (CYP) 3A4 activity was measured by ß-hydroxylation of testosterone. RESULTS: Ethyl acetate extract showed slightly higher tumor specificity (TS=2.6) and one order higher antibacterial activity (selectivity index (SI)=0.813) than other extracts (TS=1.4-2.5; SI=0.030-0.063). All extracts showed no anti-human immunodeficiency virus (HIV) activity, but some anti-herpes simplex virus (HSV) activity, which was masked by potent cytotoxicity. They showed strong inhibitory activity against CYP3A4. CONCLUSION: Ethyl acetate extraction following the removal of cytotoxic and CYP3A4 inhibitory substances by n-hexane can enhance antitumor and antibacterial activity of mastic.
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Bacterias/efectos de los fármacos , Resina Mástique/farmacología , Neoplasias/tratamiento farmacológico , Extractos Vegetales/farmacología , Antibacterianos/química , Antibacterianos/farmacología , Antivirales/química , Antivirales/farmacología , Bacterias/patogenicidad , Proliferación Celular/efectos de los fármacos , Citocromo P-450 CYP3A/genética , VIH/efectos de los fármacos , VIH/patogenicidad , Hexanos/química , Humanos , Resina Mástique/química , Neoplasias/patología , Pistacia/química , Extractos Vegetales/química , Simplexvirus/efectos de los fármacos , Simplexvirus/patogenicidadRESUMEN
Rosehip, the fruit of Rosa canina L., has traditionally been used to treat urate metabolism disorders; however, its effects on such disorders have not been characterized in detail. Therefore, the present study investigated the effects of hot water, ethanol and ethyl acetate extracts of rosehip on xanthine oxidase (XO) activity in vitro. In addition, the serum urate lowering effects of the rosehip hot water extract in a mouse model of hyperuricemia (male ddY mice, which were intraperitoneally injected with potassium oxonate) were investigated. Furthermore, the influence of rosehip hot water extract on CYP3A4 activity, which is the most important drug-metabolizing enzyme from a herb-drug interaction perspective, was investigated. Rosehip extracts of hot water, ethanol and ethyl acetate inhibited XO activity [half maximal inhibitory concentration (IC50) values: 259.6±50.6, 242.5±46.2 and 1,462.8±544.2 µg/ml, respectively]. Furthermore, the administration of 1X rosehip hot water extract significantly reduced the levels of serum urate at 8 h, which was similar when compared with the administration of 1 mg/kg allopurinol. Rosehip hot water extract only marginally affected CYP3A4 activity (IC50 value, >1 mg/ml). These findings indicate that rosehip hot water extract may present as a functional food for individuals with a high urate level, and as a therapeutic reagent for hyperuricemic patients.
RESUMEN
We previously demonstrated that tomato juice (TJ) contains potent mechanism-based inhibitor(s) of CYP3A4. In this study, we investigated the effects of TJ and grapefruit juice (GFJ) on the pharmacokinetics of the CYP3A4-substrate drugs, nifedipine (NFP) and midazolam (MDZ), in male Wistar rats. Oral administration of GFJ 90 min before the intraduodenal administration of NFP or MDZ increased the area under the concentration-time curves (AUCs) of NFP and MDZ by 32.4% and 89.4%, respectively. TJ increased MDZ blood concentrations and AUC after intraduodenal MDZ administration; however, it had no effect on NFP. When MDZ and NFP were intravenously administered, GFJ significantly increased the AUC of MDZ, but only slightly increased that of NFP. In contrast, TJ only slightly increased the AUC of MDZ. These results suggest that, similar to GFJ, TJ influences the pharmacokinetics of CYP3A4-substrate drugs; however, it may be a drug-dependent partial effect.
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Compared to studies of water extracts of plants, those utilising alkaline extracts are limited. Both water and alkaline extracts from licorice root were compared regarding their biological activities. Licorice root was successively extracted first with water or alkaline solution (pH 9 or 12), and the alkaline (pH 12.0) extract was further separated into 50% ethanol-soluble and -insoluble fractions. Viable cell number was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. Antibacterial activity against Porphyromonas gingivalis 381 was determined by turbidity assay. Cytochrome P-450 (CYP)3A4 activity was measured by ß-hydroxylation of testosterone using human recombinant CYP3A4. Radical intensity of superoxide and hydroxyl radicals was determined by electron spin resonance spectroscopy. Alkaline extraction yielded slightly higher amounts of dried materials compared to water extraction. Alkaline extract showed higher anti-HIV and antibacterial activities, and similar magnitudes of CYP3A4 inhibitory and superoxide and hydroxyl radical-scavenging activities, compared to water extract. When alkaline extract was fractionated by 50% ethanol, anti-HIV activity was recovered from the insoluble fraction representing approximately 3% of the alkaline extract, whereas antibacterial activity was concentrated in the soluble fraction rich in glycyrrhizid acid, flavanones and chalcones. All extracts and sub-fractions led to bimodal hormetic dose-response (maximum hormetic response=238%) on the bacterial growth. The present study demonstrated the superiority of alkaline extraction over water extraction for preparing anti-HIV and antibacterial agents at higher yield from licorice root.
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Glycyrrhiza/química , Extracción Líquido-Líquido/métodos , Extractos Vegetales/química , Raíces de Plantas/química , Antibacterianos/química , Antibacterianos/aislamiento & purificación , Antibacterianos/farmacología , Fármacos Anti-VIH/química , Fármacos Anti-VIH/farmacología , Cromatografía Líquida de Alta Presión , Inhibidores del Citocromo P-450 CYP3A/química , Inhibidores del Citocromo P-450 CYP3A/farmacología , Depuradores de Radicales Libres/química , Depuradores de Radicales Libres/farmacología , Concentración de Iones de Hidrógeno , Estructura Molecular , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/farmacologíaRESUMEN
We investigated the effects of α- and ß-adrenoceptor agonists on L-ascorbic acid-induced hepatocyte DNA synthesis and proliferation in primary cultures of adult rat hepatocytes. The results showed that phenylephrine (10(-6) M) and metaproterenol (10(-6) M) alone did not induce hepatocyte DNA synthesis and proliferation. However, when combined with L-ascorbic acid (10(-6) M), these adrenoceptor agonists potentiated the hepatocyte DNA synthesis and proliferation induced by L-ascorbic acid. Then intracellular signal transduction mechanisms for the effects of phenylephrine and metaproterenol on L-ascorbic acid-induced hepatocyte mitogenesis were examined. Western blot analysis showed that phenylephrine and metaproterenol did not potentiate L-ascorbic acid-induced insulin-like growth factor I receptor tyrosine kinase phosphorylation. In contrast, they both significantly potentiated L-ascorbic acid-induced extracellular-signal regulated kinase-2 (ERK2) phosphorylation within 5 min. Moreover, cell-permeable second messenger analogs phorbol ester (10(-7) M) and 8-bromo cAMP (10(-7) M) mimicked the effects of phenylephrine and metaproterenol on L-ascorbic acid-induced ERK2 phosphorylation. The effects of these adrenoceptor agents were specifically antagonized by GF109203X and H-89, respectively. These results indicate that activation of ERK2 via protein kinas C and protein kinase A represents a mechanism for potentiation of L-ascorbic acid-induced hepatocyte DNA synthesis and proliferation in primary cultures of adult rat hepatocytes.
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Ácido Ascórbico/farmacología , ADN/biosíntesis , Hepatocitos/citología , Hepatocitos/efectos de los fármacos , Receptores Adrenérgicos alfa 1/metabolismo , Receptores Adrenérgicos beta 2/metabolismo , Transducción de Señal/efectos de los fármacos , 8-Bromo Monofosfato de Adenosina Cíclica/farmacología , Agonistas de Receptores Adrenérgicos alfa 1/farmacología , Agonistas de Receptores Adrenérgicos beta 2/farmacología , Animales , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Sinergismo Farmacológico , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Hepatocitos/metabolismo , Masculino , Metaproterenol/farmacología , Fenilefrina/farmacología , Fosforilación/efectos de los fármacos , Ratas , Ratas Wistar , Receptor IGF Tipo 1/metabolismo , Acetato de Tetradecanoilforbol/farmacología , Factores de TiempoRESUMEN
BACKGROUND: We have previously reported that alkaline extract of Sasa senanensis leaves (SE) showed potent anti-HIV, anti-UV and radical scavenging activity. In the present study, we investigated the biological activities of SE-10, a granulated powder of SE supplemented with lactose, lactitol, trehalose and tea extract. MATERIALS AND METHODS: Cell viability of mock-infected, HIV-infected, and UV-irradiated cells was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. Scavenging activity of superoxide anion and hydroxyl radicals was determined by electron-spin resonance spectroscopy. Cytochrome P-450 (CYP)3A4 activity was measured by ß-hydroxylation of testosterone in human recombinant CYP3A4. RESULTS: SE-10 had slightly higher anti-HIV and anti-UV activities, but slightly lower radical-scavenging and CYP3A4-inhibitory activities, as compared with SE. CONCLUSION: The present study demonstrates that the biological activities of SE were well preserved during the manufacturing process of SE-10.
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Fármacos Anti-VIH/farmacología , Depuradores de Radicales Libres/farmacología , Extractos Vegetales/farmacología , Hojas de la Planta/química , Protectores contra Radiación/farmacología , Sasa/química , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/efectos de la radiación , Células Cultivadas , VIH-1/efectos de los fármacos , Humanos , Extractos Vegetales/química , Rayos UltravioletaRESUMEN
This study investigates whether tomato juice can inhibit cytochrome P450 (CYP) 3A4-mediated drug metabolism. Three commercially available, additive-free tomato juices, along with homogenized fresh tomato, were analyzed for their ability to inhibit testosterone 6ß-hydroxylation activity using human recombinant CYP3A4. Results were compared to that of grapefruit juice. Ethyl acetate extracts of the tomato juices moderately reduced residual activity of CYP3A4 testosterone 6ß-hydroxylation activity by 19.3-26.2% with 0-min preincubation. Residual activity was strongly reduced by 69.9-83.5% at 20-min preincubation, a reduction similar to that of grapefruit juice extract, known to contain constituents of mechanism-based inhibitors. One juice extract (tomato juice C) showed irreversible dose- and preincubation time-dependent and partial nicotinamide adenine dinucleotide phosphate (NADPH)-dependent inhibition of CYP3A4 activity. Furthermore, we examined whether the CYP3A4 inhibitory effect of tomato juice was substrate dependent by examining midazolam 1'-hydroxylation activity and nifedipine oxidation activity, in addition to testosterone 6ß-hydroxylation activity. Tomato juice showed a potent inhibitory effect on nifedipine oxidation activity, which was comparable to that on testosterone 6ß-hydroxylation activity; however, it showed a weak inhibitory effect on midazolam 1'-hydroxylation activity. We conclude that tomato juice contains one or more mechanism-based and competitive inhibitor(s) of CYP3A4. Additionally, significant CYP3A4 inhibitory activity did not result from lycopene, a major compound in tomato. Although the active compound was uncertain, a strong CYP3A4 inhibitory activity was observed in other solanaceous plants, i.e., potato, eggplant, sweet pepper, and capsicum. Therefore, responsible compounds in tomato are likely commonly shared among solanaceous vegetables.
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Bebidas , Inhibidores del Citocromo P-450 CYP3A , Solanaceae , Acetatos/química , Citocromo P-450 CYP3A/metabolismo , Humanos , NADP/metabolismo , Extractos Vegetales , Proteínas Recombinantes/antagonistas & inhibidores , Proteínas Recombinantes/metabolismo , Solanaceae/química , Solventes/química , Esteroide Hidroxilasas/antagonistas & inhibidores , Esteroide Hidroxilasas/metabolismo , UltrafiltraciónRESUMEN
BACKGROUND: We have previously reported that alkaline extract of Sasa senanensis leaves (SE) has several biological activities characteristic of lignin-carbohydrate complex (LCC). In the present study, we compared the biological activity of three commercially available products of SE (products A, B and C). MATERIALS AND METHODS: Cell viability of mock-infected, HIV-infected, UV-irradiated cells was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. Radical intensity was determined by electron spin resonance spectroscopy. Cytochrome P-450 (CYP)3A4 activity was measured by ß-hydroxylation of testosterone in human recombinant CYP3A4. RESULTS: Product A is a pure SE that contains Fe(II)-chlorophyllin, whereas products B and C contain Cu(II)-chlorophyllin and less LCC. Product C is supplemented with ginseng and pine (Pinus densiflora) leaf extracts. Product A exhibited 5-fold higher anti-HIV, 4-fold higher anti-UV, 5-fold higher hydroxyl radical-scavenging, and 3-fold lower CYP3A4 inhibitory activities as compared to those of product B, and 5-fold higher, 1.5-fold higher, comparable, and 7-fold lower activities, respectively, as compared to those of product C. CONCLUSION: The present study demonstrates for the first time the superiority of product A over products B and C, suggesting the beneficial role of LCC and Fe(II)-chlorophyllin.
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Fármacos Anti-VIH/farmacología , Depuradores de Radicales Libres/farmacología , Extractos Vegetales/farmacología , Protectores contra Radiación/farmacología , Sasa/química , Linfocitos T/efectos de los fármacos , Fármacos Anti-VIH/aislamiento & purificación , Fármacos Anti-VIH/toxicidad , Carcinoma de Células Escamosas/patología , Línea Celular Tumoral/efectos de los fármacos , Línea Celular Tumoral/efectos de la radiación , Línea Celular Tumoral/virología , Supervivencia Celular , Clorofilidas/análisis , Clorofilidas/farmacología , Citocromo P-450 CYP3A , Inhibidores del Citocromo P-450 CYP3A , Combinación de Medicamentos , Espectroscopía de Resonancia por Spin del Electrón , Inhibidores Enzimáticos/aislamiento & purificación , Inhibidores Enzimáticos/farmacología , Inhibidores Enzimáticos/toxicidad , Depuradores de Radicales Libres/aislamiento & purificación , Depuradores de Radicales Libres/toxicidad , VIH-1 , Virus Linfotrópico T Tipo 1 Humano , Humanos , Lignina/farmacología , Lignina/toxicidad , Neoplasias de la Boca/patología , Medicamentos sin Prescripción , Panax/química , Pinus/química , Extractos Vegetales/toxicidad , Hojas de la Planta/química , Protectores contra Radiación/aislamiento & purificación , Protectores contra Radiación/toxicidad , Proteínas Recombinantes/antagonistas & inhibidores , Linfocitos T/virología , Rayos UltravioletaRESUMEN
The administration of fibrates (fenofibrate, bezafibrate and clofibric acid) to rats induced stearoyl-CoA desaturase (SCD) in the liver, and increased relative expression of mRNAs encoding SCD1 and SCD2 in dose- and time-dependent manners. The magnitudes of the increases in SCD2 mRNA level caused by fenofibrate and clofibric acid were much higher than those of SCD1 at relatively higher doses of the fibrates, and a relatively long time (7 or 14 d) was required for significant induction of SCD2 mRNA expression compared with that of SCD1. Although the absolute number of transcripts for SCD2 was 1,800 times lower than that of SCD1 in the control liver, it was strikingly increased by fibrates. These results suggest that differential regulations operate for the gene expression between SCD1 and SCD2, and that the physiological significance of SCD2 is distinct from that of SCD1 in the liver.
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Bezafibrato/farmacología , Ácido Clofíbrico/farmacología , Activadores de Enzimas/farmacología , Fenofibrato/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Hígado/enzimología , Estearoil-CoA Desaturasa/metabolismo , Animales , Relación Dosis-Respuesta a Droga , Activación Enzimática/efectos de los fármacos , Hígado/efectos de los fármacos , Masculino , ARN Mensajero/metabolismo , Ratas , Ratas Wistar , Estearoil-CoA Desaturasa/genéticaRESUMEN
The effects of 2-(4-chlorophenoxy)-2-methylpropionic acid (clofibric acid) on the formation of oleic acid (18:1) from stearic acid (18:0) and utilization of the 18:1 formed for phosphatidylcholine (PC) formation in endoplasmic reticulum in the liver of rats were studied in vivo. [¹4C]18:0 was intravenously injected into control Wistar male rats and rats that had been fed on a diet containing 0.5% (w/w) clofibric acid for 7 days; and the distribution of radiolabeled fatty acids among subcellular organelles, microsomes, peroxisomes, and mitochondria, was estimated on the basis of correction utilizing the yields from homogenates of marker enzymes for these organelles. The radioactivity was mostly localized in microsomes and the radiolabeled fatty acids present in microsomes were significantly increased by the treatment of rats with clofibric acid. The formation of radiolabeled 18:1 in microsomes markedly increased and incorporations of the formed [¹4C]18:1 into PC and phosphatidylethanolamine in microsomes were augmented in response to clofibric acid. The [¹4C]18:1 incorporated into PC was mostly located at the C-2 position, but not the C-1 position, of PC, and the radioactivity in 18:1 at the C-2 position of PC was strikingly increased by clofibric acid. These results obtained from the in vivo experiments directly link the findings that clofibric acid treatment induces microsomal stearoyl-CoA desaturase and 1-acylglycerophosphocholine acyltransferase in the liver and the findings that the treatment with the drug elevated absolute mass and mass proportion of 18:1 at the C-2 position, but not the C-1 position, of PC in the liver together.
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Ácido Clofíbrico/farmacología , Retículo Endoplásmico/efectos de los fármacos , Retículo Endoplásmico/metabolismo , Hígado/metabolismo , Microsomas Hepáticos/efectos de los fármacos , Microsomas Hepáticos/metabolismo , Ácido Oléico/biosíntesis , 1-Acilglicerofosfocolina O-Aciltransferasa/metabolismo , Alimentación Animal , Animales , Ácido Clofíbrico/metabolismo , Ácidos Grasos/metabolismo , Masculino , Fosfatidilcolinas/biosíntesis , Ratas , Ratas Wistar , Ácidos Esteáricos/metabolismo , Ácidos Esteáricos/farmacología , Estearoil-CoA Desaturasa/metabolismoRESUMEN
Alterations by perfluorinated fatty acids (PFCAs) with a chain length of 6-9 carbons in the fatty acid profile of hepatic lipids of mice were investigated. The characteristic changes caused by all the PFCAs examined were increases in the contents and proportions of oleic acid (18 : 1), palmitoleic acid (16 : 1) and 8,11,14-eicosatrienoic acid (20 : 3) in hepatic lipids. Hepatic contents of palmitic acid were also increased by the treatments with the PFCAs. These effects were almost dependent on the hepatic concentrations of PFCA molecules regardless of their carbon chain length. Perfluorooctanoic acid elevated the expressions of mRNA encoding acetyl-CoA carboxylase, fatty acid synthase, malic enzyme, stearoyl-CoA desaturase (SCD) (SCD1 and 2), chain elongase (ELOVL5), Δ6 desaturase (Fads2), 1-acylglycerophosphocholine acyltransferase (LPCAT) (LPCAT3). The four PFCAs examined induced microsomal SCD and LPCAT in hepatic concentration-dependent manners regardless of carbon chain length. One linear regression line was confirmed between LPCAT activity and hepatic concentration of PFCA at wide range of the concentration, whereas the induction of SCD was saturable at relatively low concentration of PFCAs. These results suggest (i) that PFCAs with a chain length of 6-9 carbons change the fatty acid profile of hepatic lipids by increasing contents and proportions of 16 : 1, 18 : 1 and 20 : 3, (ii) that these alterations in fatty acid profile are caused by up-regulation of SCD, de novo fatty acid synthesis, chain elongase and Δ6 desaturase and (iii) that the mechanism underlying SCD induction is, in part, mediated through peroxisome proliferator-activated receptor α.
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Contaminantes Ambientales/toxicidad , Ácidos Grasos/análisis , Ácidos Grasos/toxicidad , Fluorocarburos/toxicidad , Hepatomegalia/inducido químicamente , Hígado/química , Hígado/efectos de los fármacos , Ácido 8,11,14-Eicosatrienoico/análisis , Acetiltransferasas/genética , Acetiltransferasas/metabolismo , Animales , Caprilatos/análisis , Caprilatos/toxicidad , Relación Dosis-Respuesta a Droga , Contaminantes Ambientales/análisis , Contaminantes Ambientales/química , Elongasas de Ácidos Grasos , Ácidos Grasos/biosíntesis , Ácidos Grasos/química , Ácidos Grasos Monoinsaturados/análisis , Fluorocarburos/análisis , Fluorocarburos/química , Regulación Enzimológica de la Expresión Génica , Hepatomegalia/metabolismo , Isoenzimas/genética , Isoenzimas/metabolismo , Linoleoil-CoA Desaturasa/genética , Linoleoil-CoA Desaturasa/metabolismo , Hígado/metabolismo , Masculino , Ratones , Ratones Endogámicos , Peso Molecular , Ácido Oléico/análisis , PPAR alfa/metabolismo , ARN Mensajero/metabolismo , Estearoil-CoA Desaturasa/genética , Estearoil-CoA Desaturasa/metabolismoRESUMEN
Cobalt focus is a seizure focus model in which cerebral neurons exhibit long-lasting severe spike discharges, followed by neuronal death. However, the neuronal death is prevented when peony root extract (PR) is administered prior to cobalt application. We tested the hypothesis that PR modulates the expression of neuroprotective proteins in the cerebrum of mouse cobalt focus by proteomic analysis using two-dimensional polyacrylamide gel electrophoresis and mass spectrometry to screen for differentially expressed proteins. Analyses revealed that transthyretin, a carrier protein for thyroid hormones and retinoids, and the brain form of phosphoglycerate mutase, a glycolytic enzyme, were upregulated in the cobalt-treated mouse cerebrum and further increased by PR administration in association with upregulation of neurogranin/RC3, a target of the transcriptional activation by thyroid hormones and retinoids. These findings suggest that PR-induced protection of mouse cerebral neurons involves neurotrophic events caused by thyroid hormones and/or retinoids and enhanced glycolysis.
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Anticonvulsivantes/administración & dosificación , Cerebro/efectos de los fármacos , Paeonia , Fosfoglicerato Mutasa/metabolismo , Extractos Vegetales/administración & dosificación , Raíces de Plantas , Prealbúmina/metabolismo , Convulsiones/prevención & control , Animales , Cerebro/metabolismo , Cobalto/antagonistas & inhibidores , Cobalto/toxicidad , Modelos Animales de Enfermedad , Electroforesis en Gel Bidimensional , Ratones , Ratones Endogámicos C57BL , Neurogranina/análisis , Neurogranina/metabolismo , Fosfoglicerato Mutasa/análisis , Prealbúmina/análisis , Proteómica , Convulsiones/inducido químicamente , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Hormonas Tiroideas/metabolismo , Regulación hacia ArribaRESUMEN
To elucidate the mechanism of inhibitory action of peony root extract on pentylenetetrazol-induced bursting activity, effects of peony root extract on the iberiotoxin-sensitive large conductance calcium-activated potassium (BKCa) current that plays an essential role in the production of bursting activity were investigated. Peony root extract showed a clear inhibitory effect on the iberiotoxin-sensitive calcium-activated potassium current. Peony root extract also showed clear inhibitory effects on spontaneous bursting activity and BKCa current in the cerebral cortical neurons of the EL mouse, a hereditary epilepsy animal model. These results together with our previous studies, including the protective effect against neuron damage, indicate that peony root extract is a promising herbal drug for inhibition of convulsions.
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Potenciales de Acción/efectos de los fármacos , Anticonvulsivantes/farmacología , Paeonia , Fitoterapia , Extractos Vegetales/farmacología , Canales de Potasio Calcio-Activados/efectos de los fármacos , Animales , Anticonvulsivantes/administración & dosificación , Anticonvulsivantes/uso terapéutico , Células Cultivadas , Corteza Cerebral/citología , Corteza Cerebral/embriología , Ratones , Ratones Endogámicos , Extractos Vegetales/administración & dosificación , Extractos Vegetales/uso terapéutico , Raíces de Plantas , Convulsiones/tratamiento farmacológicoRESUMEN
Doxorubicin (adriamycin), an anthracycline antibiotic, showed higher cytotoxic activity against human tumor cell lines (oral squamous cell carcinoma HSC-2, HSC-3, submandibular gland carcinoma HSG, promyelocytic leukemia HL-60) than against normal human cells (gingival fibroblast HGF, pulp cell HPC, periodontal ligament fibroblast HPLF). Doxorubicin activated caspases 3, 8 and 9 in both HSC-2 and HL-60 cells, but induced internucleosomal DNA fragmentation only in HL-60 cells. Western blot analysis showed that doxorubicin did not significantly change the intracellular concentration of Bcl-2, Bax and Bad in HL-60 cells. Real-time PCR analysis showed that HPC cells expressed the highest amount of mdr1 mRNA, followed by HSC-2 > HGF > HSC-3 > HPLF > HSG > HL-60. ESR spectroscopy showed that doxorubicin produced no discernible radical under alkaline conditions (pH 7.4 to 10.5) except at pH 12.5, and it did not scavenge O2-, NO and DPPH radicals. The present study demonstrates that doxorubicin induces the tumor-specific cytotoxicity and some, but not all, apoptosis markers possibly by a radical-independent mechanism, and that mdr1 expression in the tumor cells is not related to the tumor specificity of doxorubicin.
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Antibióticos Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Doxorrubicina/farmacología , Antibióticos Antineoplásicos/metabolismo , Carcinoma de Células Escamosas/tratamiento farmacológico , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patología , Línea Celular Tumoral , Doxorrubicina/metabolismo , Espectroscopía de Resonancia por Spin del Electrón , Depuradores de Radicales Libres/farmacología , Células HL-60 , Humanos , Neoplasias de la Boca/tratamiento farmacológico , Neoplasias de la Boca/metabolismo , Neoplasias de la Boca/patología , Superóxidos/metabolismoRESUMEN
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) has long been recognized as a classical glycolytic protein; however, previous studies by our group and others have demonstrated that GAPDH is a general mediator initiating one or more apoptotic cascades. Our most recent findings have elucidated that an expression of a pro-apoptotic protein GAPDH is critically regulated at the promoter region of the gene. Apoptotic signals for its subsequent aggregate formation and nuclear translocation are controlled by the respective functional domains harboured within its cDNA component. In this study, coexpression of GAPDH with either wild-type or mutant (A53T) alpha-synuclein and less likely with beta-synuclein in transfected COS-7 cells was found to induce Lewy body-like cytoplasmic inclusions. Unlike its full-length construct, the deleted mutant GAPDH construct (C66) abolished these apoptotic signals, disfavouring the formation of inclusions. The generated inclusions were ubiquitin- and thioflavin S-positive appearing fibrils. Furthermore, GAPDH coimmunoprecipitated with wild-type alpha-synuclein in this paradigm. Importantly, immunohistochemical examinations of post mortem materials from patients with sporadic Parkinson's disease revealed the colocalized profiles immunoreactive against these two proteins in the peripheral zone of Lewy bodies from the affected brain regions (i.e. locus coeruleus). Moreover, a quantitative assessment showed that about 20% of Lewy bodies displayed both antigenicities. These results suggest that pro-apoptotic protein GAPDH may be involved in the Lewy body formation in vivo, probably associated with the apoptotic death pathway.