RESUMEN
Selective estrogen receptor modulators (SERMs) are a class of therapeutic chemicals which present tissue-specific estrogen receptor modulating activity. Neonatal exposure to SERMs has been reported to adversely affect central nervous system development, however, mechanism and involvement of hypothalamic kisspeptin neurone in this impairment remains undetermined. To clarify this uncertainty, neonates from female Donryu rats were subcutaneously injected with raloxifene (RLX) at 0.1, 1, and 10mg/kg or tamoxifen (TMX) at 10mg/kg on postnatal day 0, and then hypothalamic KiSS1 mRNA expression and gonadotropin levels were investigated during young adulthood and estrous cycling was monitored until middle age. Treatment with RLX or TMX at 10mg/kg significantly depressed luteinizing hormone surge levels and KiSS1 mRNA expression in the anteroventral periventricular nucleus (AVPV), the control center of estrous cyclicity. The 10mg/kg TMX group also showed decreased levels of follicle-stimulating hormone and KiSS1 mRNA expression in the arcuate nucleus (ARC). Early cessation of normal estrous cycling was observed in the 10mg/kg RLX group, while the estrous cycle in the 10mg/kg TMX group had ceased by the start of the analysis. The same dose of tamoxifen or raloxifene had either weak-estrogenic or anti-estrogenic activity on the uterus, respectively; however, treatment in adulthood with both SERMs did not affect KiSS1 mRNA expression in either the AVPV or ARC in the present study. These results indicate that neonatal exposure to SERMs could disrupt neuroendocrine development and postnatal reproductive function through the alteration of kisspeptin neurons.
Asunto(s)
Discapacidades del Desarrollo/inducido químicamente , Enfermedades del Sistema Endocrino/inducido químicamente , Hipotálamo/patología , Kisspeptinas/metabolismo , Neuronas/metabolismo , Moduladores Selectivos de los Receptores de Estrógeno/toxicidad , Factores de Edad , Animales , Animales Recién Nacidos , Peso Corporal/efectos de los fármacos , Discapacidades del Desarrollo/patología , Modelos Animales de Enfermedad , Enfermedades del Sistema Endocrino/patología , Estradiol/análogos & derivados , Estradiol/farmacología , Ciclo Estral/efectos de los fármacos , Femenino , Hormonas/metabolismo , Hipotálamo/efectos de los fármacos , Kisspeptinas/genética , Neuronas/efectos de los fármacos , Ovariectomía , Embarazo , Progesterona/farmacología , Clorhidrato de Raloxifeno/farmacología , Ratas , Tamoxifeno/farmacologíaRESUMEN
Nuclear receptors play important roles in chemically induced liver hypertrophy in rodents. To clarify the involvement of constitutive androstane receptor (CAR) and other nuclear receptors in mouse liver hypertrophy induced by different doses of piperonyl butoxide (PBO), wild-type and CAR-knockout mice were administered PBO (200, 1,000, or 5,000 ppm) in the basal diet for 1 week. Increased liver weight and diffuse hepatocellular hypertrophy were observed at 5,000 ppm for both genotypes, accompanied by increased Cyp3a11 mRNA and CYP3A protein expression, suggesting that CAR-independent pathway, possibly pregnane X receptor (PXR), plays a major role in the induction of hypertrophy. Moreover, wild-type mice at 5,000 ppm showed enhanced hepatocellular hypertrophy and strong positive staining for CYP2B in the centrilobular area, suggesting the localized contribution of CAR. At 1,000 ppm, only wild-type mice showed liver weight increase and centrilobular hepatocellular hypertrophy concurrent with elevated Cyp2b10 mRNA expression and strong CYP2B staining, indicating that CAR was essential at 1,000 ppm. We concluded that high-dose PBO induced hypertrophy via CAR and another pathway, while lower dose of PBO induced a pathway mediated predominantly by CAR. The dose-responsiveness on liver hypertrophy is important for understanding the involvement of nuclear receptors.
Asunto(s)
Hepatomegalia/inducido químicamente , Butóxido de Piperonilo/efectos adversos , Receptores Citoplasmáticos y Nucleares/fisiología , Animales , Hidrocarburo de Aril Hidroxilasas/genética , Hidrocarburo de Aril Hidroxilasas/metabolismo , Citocromo P-450 CYP3A/genética , Citocromo P-450 CYP3A/metabolismo , Sistema Enzimático del Citocromo P-450/genética , Sistema Enzimático del Citocromo P-450/metabolismo , Familia 2 del Citocromo P450 , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Expresión Génica , Hepatomegalia/genética , Hepatomegalia/patología , Hipertrofia , Hígado/metabolismo , Hígado/patología , Masculino , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Ratones Endogámicos C3H , Tamaño de los Órganos/efectos de los fármacos , Receptor X de Pregnano , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptores Sensibles al Calcio , Receptores Acoplados a Proteínas G/fisiología , Receptores de Esteroides/genética , Receptores de Esteroides/fisiología , Transducción de Señal/genética , Transducción de Señal/fisiología , Esteroide Hidroxilasas/genética , Esteroide Hidroxilasas/metabolismoRESUMEN
Neonatal exposure to estrogens is known to cause delayed effects, a late-occurring adverse effect on adult female reproductive functions, such as early onset of age-matched abnormal estrous cycling. However, the critical period in which neonates are sensitive to delayed effects inducible by exogenous estrogen exposure has not been clearly identified. To clarify this window, we examined the intensity and timing of delayed effects using rats exposed to ethynylestradiol (EE) at various postnatal ages. After subcutaneous administration of a single dose of EE (20 µg/kg, which induces delayed effects) on Postnatal Day (PND) 0, 5, 10, or 14 in Wistar rats, hypothalamic and hormonal alterations in young adults and long-term estrous cycling status were investigated as indicators of delayed effects. In young adults, peak luteinizing hormone concentrations at the time of the luteinizing hormone surge showed a decreasing trend, and KiSS1 mRNA expression of the anterior hypothalamus and number of KiSS1-positive cells in the anteroventral periventricular nucleus were significantly decreased in the PND 0, 5, and 10 groups. The reduction in KiSS1 mRNA and KiSS1-postive cells was inversely correlated with age at time of exposure. These groups also exhibited early onset of abnormal estrous cycling, starting from 17 wk of age in the PND0 group and 19 wk of age in the PND5 and 10 groups. These indicators were not apparent in the PND14 group. Our results suggest that PND0-PND10 is the critical window of susceptibility for delayed effects, and PND14 is presumed to be the provisional endpoint of the window.
Asunto(s)
Etinilestradiol/toxicidad , Envejecimiento , Animales , Animales Recién Nacidos , Peso Corporal/efectos de los fármacos , Ciclo Estral/efectos de los fármacos , Etinilestradiol/sangre , Femenino , Hormona Folículo Estimulante/sangre , Hipotálamo/efectos de los fármacos , Hipotálamo/crecimiento & desarrollo , Hipotálamo Anterior/metabolismo , Kisspeptinas/biosíntesis , Kisspeptinas/genética , Hormona Luteinizante/sangre , Embarazo , Efectos Tardíos de la Exposición Prenatal , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Ratas , Ratas Wistar , Diferenciación Sexual/efectos de los fármacos , Vagina/efectos de los fármacos , Vagina/crecimiento & desarrollo , Enfermedades Vaginales/inducido químicamente , Enfermedades Vaginales/patologíaRESUMEN
Ginkgo biloba extract (GBE) is commonly used as a herbal supplement. The National Toxicology Program (NTP) study of GBE reported clear evidence of hepatocarcinogenicity in mice. To clarify the mode of action (MOA) for hepatocarcinogenesis by GBE, we investigated the involvement of the constitutive androstane receptor (CAR) in hepatocarcinogenesis induced by GBE using CAR-knockout (CARKO) and wild type (WT) mice. We used the same lot of GBE that was used for the NTP study. In 1-week GBE dietary treatment, hepatocellular DNA replication was increased in WT mice but not in CARKO mice. In 4- or 13-week treatment, greater hepatic Cyp2b10 induction and hepatocellular hypertrophy were observed in WT mice, whereas these effects of GBE were much smaller in CARKO mice. In a two-stage hepatocarcinogenesis model initiated by diethylnitrosamine, 27-week treatment with GBE resulted in an increase of eosinophilic altered foci and adenomas in WT mice. By contrast, foci and adenomas were clearly less evident in CARKO mice. These results indicate that GBE-induced hepatocarcinogenesis is mainly CAR-mediated. Since CAR-mediated MOA for hepatocarcinogenesis in rodents is considered to be qualitatively implausible for humans, our findings would be helpful to evaluate the carcinogenic characterization of GBE to humans.
Asunto(s)
Cocarcinogénesis/metabolismo , Suplementos Dietéticos/efectos adversos , Ginkgo biloba/química , Hepatomegalia/etiología , Neoplasias Hepáticas/etiología , Extractos Vegetales/efectos adversos , Receptores Citoplasmáticos y Nucleares/agonistas , Adenoma de Células Hepáticas/inducido químicamente , Adenoma de Células Hepáticas/etiología , Adenoma de Células Hepáticas/metabolismo , Adenoma de Células Hepáticas/patología , Animales , Hidrocarburo de Aril Hidroxilasas/química , Hidrocarburo de Aril Hidroxilasas/genética , Hidrocarburo de Aril Hidroxilasas/metabolismo , Carcinógenos/química , Carcinógenos/toxicidad , Cocarcinogénesis/patología , Receptor de Androstano Constitutivo , Inductores de las Enzimas del Citocromo P-450/efectos adversos , Familia 2 del Citocromo P450 , Replicación del ADN , Dietilnitrosamina/agonistas , Dietilnitrosamina/toxicidad , Hepatomegalia/metabolismo , Hepatomegalia/patología , Japón , Hígado/efectos de los fármacos , Hígado/metabolismo , Hígado/patología , Neoplasias Hepáticas/inducido químicamente , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patología , Masculino , Ratones Endogámicos C3H , Ratones Noqueados , Receptores Citoplasmáticos y Nucleares/genética , Receptores Citoplasmáticos y Nucleares/metabolismo , Esteroide Hidroxilasas/química , Esteroide Hidroxilasas/genética , Esteroide Hidroxilasas/metabolismo , Pruebas de Toxicidad SubcrónicaRESUMEN
We previously reported that neonatal exposure to 17α-ethynylestradiol (EE) led to delayed adverse effects in which age-related anovulation after sexual maturation was accelerated. To identify early indicators of these adverse effects, female Wistar Hannover GALAS rats received a single EE injection (0, 0.02, 0.2, 2, 20, or 200 µg/kg) within 24 hr of birth. Histopathological changes in ovarian and uterine development were investigated from postnatal day (PND) 14 to 10 weeks of age. Immunohistochemical expression of estrogen receptor alpha (ERα) in the uterus, serum levels of sex-related hormones and gene expression in the hypothalamus were examined. Although neonatal exposure to EE did not affect body growth or ovarian development, serum FSH tended to decrease at doses ≥ 2 µg/kg, and Kiss1 mRNA level in the whole hypothalamus was significantly decreased in all EE-treated groups at PND14.The number of uterine glands at PND21 was suppressed at doses ≥ 20 µg/kg, and ERα expression in the uterine epithelium at estrus stage decreased in a dose-dependent manner at 10 weeks of age. These results demonstrated that the various identified changes that occurred before the appearance of delayed adverse effects could be candidate early indicators.
Asunto(s)
Etinilestradiol/efectos adversos , Ovario/efectos de los fármacos , Útero/efectos de los fármacos , Animales , Animales Recién Nacidos , Anovulación/inducido químicamente , Relación Dosis-Respuesta a Droga , Receptor alfa de Estrógeno/metabolismo , Etinilestradiol/administración & dosificación , Femenino , Hormona Folículo Estimulante/sangre , Hipotálamo/metabolismo , Kisspeptinas/metabolismo , Ratas , Ratas Wistar , Maduración Sexual/efectos de los fármacos , Útero/metabolismoRESUMEN
The National Toxicology Program study of Ginkgo biloba extract (GBE), a herbal supplement, reported concerns regarding genotoxicity and clear evidence of hepatocarcinogenicity and liver hypertrophy in mice. To clarify the genotoxicity of GBE in vivo, we performed reporter gene mutation assay using gpt delta mice. We also used a combined liver comet assay and bone marrow micronucleus assay using C3H-derived constitutive androstane receptor knockout (CARKO) and wild-type mice. No remarkable increases in gpt or Spi(-) mutation frequencies were observed in DNA extracted from the livers of gpt delta mice that had been exposed to GBE up to 2000 mg/kg bw/day. In the comet and micronucleus assays, no statistically significant increases in positive cells were observed at doses up to 2000 mg/kg bw/day of GBE in either mouse genotype. The present study provides clear evidence that GBE is not genotoxic in vivo. Our results indicate that GBE-induced hepatocarcinogenesis in mice occurs through a non-genotoxic mode of action.
Asunto(s)
Ginkgo biloba/química , Mutágenos/toxicidad , Extractos Vegetales/toxicidad , Receptores Citoplasmáticos y Nucleares/genética , Transferasas (Grupos de Otros Fosfatos Sustitutos)/genética , Animales , Peso Corporal/efectos de los fármacos , Ensayo Cometa , Receptor de Androstano Constitutivo , Femenino , Hígado/efectos de los fármacos , Masculino , Ratones , Ratones Noqueados , Pruebas de Micronúcleos , Tamaño de los Órganos/efectos de los fármacosRESUMEN
In a 13-week feeding toxicity study of grape skin extract (GSE) performed previously, 5.0% GSE showed diffuse hypertrophy and basophilia in rat parotid glands. To clarify whether the change in the parotid glands was an adverse effect of GSE, 6-week-old male F344 rats were fed a diet containing 5.0% GSE or were administered a dose corresponding to the dietary concentration via gavage for 4 weeks, and the treatment was stopped for 2 weeks. To ascertain the effect of astringency, other animals were fed a diet containing 5.0% tannic acid (TA) using the same protocol as the GSE feed group. Control groups were fed a basal diet or were administered sterilized distilled water by gavage. In the GSE and TA feed groups, diffuse severe hypertrophy and basophilia in the parotid glandular epithelial cells were observed. Macroscopic, microscopic, and ultrastructural characteristics consistent with cellular hypertrophy was less apparent after the recovery period in both feed groups. In contrast, no changes were observed in the parotid glands of the gavage GSE and control groups at week 4. Based on these findings of parotid hypertrophy without cytotoxicity, the data from this and previous studies suggest that hypertrophy of the parotid glands induced by feeding treatment with GSE is an adaptive non-adverse effect that is reversible upon removal of the sialotrophic agent.
Asunto(s)
Glándula Parótida/efectos de los fármacos , Extractos Vegetales/farmacología , Vitis/química , Adaptación Biológica/efectos de los fármacos , Administración Oral , Animales , Peso Corporal/efectos de los fármacos , Hipertrofia , Masculino , Enfermedades de las Parótidas/inducido químicamente , Enfermedades de las Parótidas/patología , Glándula Parótida/citología , Glándula Parótida/patología , RatasRESUMEN
A 13-week repeated oral dose toxicity study of grape skin extract (GSE) was performed using F344 rats. Four groups of animals, each consisting of ten males and ten females, were fed a diet containing 0%, 0.2%, 1.0% or 5.0% GSE for 13 weeks. Throughout the experiment, there were no treatment-related changes in clinical signs, body weight or mean food intake in any of the treated groups of either gender. Hematological studies and serum biochemical analyses revealed no treatment-related changes in all groups in both genders. In the glandular epithelial cells of the parotid glands, diffuse hypertrophy and basophilia was observed in all animals in both 5.0% groups. Hypertrophy of the parotid glands was not detected in the 0.2% or the 1.0% dose groups. In female kidneys, slight calcification in the renal proximal tubules of the cortex and medulla was observed in all groups including controls. This is a common spontaneous change in female rats, and the incidence was comparable between controls and treated groups. However, the number of tubules with calcification was higher in the 5.0% group based on a semi-morphometric analysis. Based on the histopathology of the parotid glands and the minor change in the kidneys, the no observed adverse effect level (NOAEL) of GSE in the present study was a 1.0% treatment dose in both genders (males: 0.6 ± 0.2 g/kg body weight/day; females: 0.7 ± 0.1 g/kg body weight/day).
Asunto(s)
Extractos Vegetales/toxicidad , Vitis/química , Administración Oftálmica , Animales , Peso Corporal/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Ingestión de Alimentos/efectos de los fármacos , Células Epiteliales/efectos de los fármacos , Células Epiteliales/patología , Femenino , Riñón/efectos de los fármacos , Masculino , Nivel sin Efectos Adversos Observados , Glándula Parótida/citología , Extractos Vegetales/administración & dosificación , Ratas , Ratas Endogámicas F344 , Factores de Tiempo , Pruebas de Toxicidad Subcrónica , Vitis/anatomía & histologíaRESUMEN
Lucidin-3-O-primeveroside (LuP) is a component of madder color (MC), a compound which is carcinogenic in the kidney and liver of rats. Since LuP is metabolized to generate genotoxic compounds such as lucidin (Luc) and rubiadin, it is likely that these play key roles in MC carcinogenesis. In fact, after incubation of Luc with calf thymus DNA, Luc-N(2)-dG and N(6)-dA adducts were reportedly formed, possibly via the sulfotransferase metabolic pathway. However, the precise extent of formation in vivo remains uncertain. In the present study, to quantitatively determine Luc-specific DNA adducts in in vivo samples, we developed an online sample purification method using column-switching and an isotope dilution LC-ESI-MS/MS technique. The limits of quantification were 0.2 and 0.04 fmol on column for Luc-N(2)-dG and N(6)-dA adducts, respectively. Using the new analytical method, we attempted to measure adduct levels in the kidneys and livers of rats treated with 0.06, 0.3, and 1.5% LuP in the diet for one week. Luc-N(2)-dG and N(6)-dA adducts in these organs were detected at ranges from 7.97 to 51.67/10(9) dG and from1.83 to 37.10/10(9) dA, respectively. Dose-dependent increases of each adduct were observed in both organs. These quantitative data obtained with our newly developed analytical method might help to improve our understanding of MC carcinogenesis.
Asunto(s)
Antraquinonas/análisis , Carcinógenos/análisis , Aductos de ADN/análisis , Riñón/metabolismo , Hígado/metabolismo , Extractos Vegetales/química , Rubia/química , Espectrometría de Masa por Ionización de Electrospray/métodos , Animales , Antraquinonas/metabolismo , Carcinógenos/metabolismo , Cromatografía Líquida de Alta Presión/métodos , Aductos de ADN/metabolismo , Límite de Detección , Masculino , Extractos Vegetales/metabolismo , Ratas , Ratas Endogámicas F344 , Rubia/metabolismo , Espectrometría de Masas en Tándem/métodosRESUMEN
BACKGROUND: Glaucoma is a leading cause of blindness worldwide. Nonetheless, the mechanism of its pathogenesis has not been well-elucidated, particularly at the molecular level, because of insufficient availability of experimental genetic animal models. METHODOLOGY/PRINCIPAL FINDINGS: Here we demonstrate that deficiency of Vav2 and Vav3, guanine nucleotides exchange factors for Rho guanosine triphosphatases, leads to an ocular phenotype similar to human glaucoma. Vav2/Vav3-deficient mice, and to a lesser degree Vav2-deficient mice, show early onset of iridocorneal angle changes and elevated intraocular pressure, with subsequent selective loss of retinal ganglion cells and optic nerve head cupping, which are the hallmarks of glaucoma. The expression of Vav2 and Vav3 tissues was demonstrated in the iridocorneal angle and retina in both mouse and human eyes. In addition, a genome-wide association study screening glaucoma susceptibility loci using single nucleotide polymorphisms analysis identified VAV2 and VAV3 as candidates for associated genes in Japanese open-angle glaucoma patients. CONCLUSIONS/SIGNIFICANCE: Vav2/Vav3-deficient mice should serve not only as a useful murine model of spontaneous glaucoma, but may also provide a valuable tool in understanding of the pathogenesis of glaucoma in humans, particularly the determinants of altered aqueous outflow and subsequent elevated intraocular pressure.
Asunto(s)
Predisposición Genética a la Enfermedad , Glaucoma/genética , Proteínas Proto-Oncogénicas c-vav/genética , Animales , Antihipertensivos/uso terapéutico , Pueblo Asiatico/genética , Ojo/metabolismo , Ojo/patología , Glaucoma de Ángulo Abierto/etnología , Glaucoma de Ángulo Abierto/genética , Humanos , Hidroftalmía/genética , Hidroftalmía/metabolismo , Inmunohistoquímica , Hibridación in Situ , Presión Intraocular/efectos de los fármacos , Japón , Latanoprost , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Degeneración Nerviosa , Disco Óptico/metabolismo , Disco Óptico/patología , Polimorfismo de Nucleótido Simple , Prostaglandinas F Sintéticas/uso terapéutico , Proteínas Proto-Oncogénicas c-vav/deficiencia , Proteínas Proto-Oncogénicas c-vav/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa InversaAsunto(s)
Suplementos Dietéticos/análisis , Aditivos Alimentarios/análisis , Análisis de los Alimentos , Contaminación de Alimentos/análisis , Inocuidad de los Alimentos/métodos , Alimentos Orgánicos/análisis , Pruebas de Toxicidad , Animales , Suplementos Dietéticos/toxicidad , Aditivos Alimentarios/toxicidad , Embalaje de Alimentos , Alimentos Orgánicos/toxicidad , Humanos , Ratones , Residuos de Plaguicidas/análisis , Residuos de Plaguicidas/toxicidad , Juego e Implementos de Juego , Ratas , Medición de RiesgoRESUMEN
Madder color (MC), a food coloring extracted from roots of Rubia tinctorum L., has been proven to exert carcinogenicity in the rat kidney and liver. Furthermore, it induces DNA adducts in the kidney, liver, and colon. MC is in fact composed of anthraquinones such as lucidin-3-O-primeveroside and alizarin. To clarify which of these might be responsible for the carcinogenicity, a rat medium-term multi-organ carcinogenesis bioassay was performed focusing on the kidney, liver, and colon. Male 6-week-old F344 rats after receiving five different carcinogens were fed a diet containing either 0.008% or 0.04% of alizarin or rubiadin, a metabolite of lucidin-3-O-primeveroside, for 23 weeks. Treatment with 0.04% rubiadin significantly increased atypical renal tubules/hyperplasias and induced renal cell adenomas and carcinomas. Renal cell tumors were also increased with 0.04% alizarin, although at lower incidence than with rubiadin. In addition, glutathione S-transferase placental form-positive liver cell foci and large intestinal dysplasias were significantly increased with 0.04% rubiadin. These results indicate that both rubiadin and alizarin can increase renal preneoplastic lesions, the potential of the latter being weaker. Rubiadin may also target the liver and large intestine, suggesting a major role in madder color-induced carcinogenicity.
Asunto(s)
Antraquinonas/toxicidad , Neoplasias del Colon/inducido químicamente , Neoplasias Renales/inducido químicamente , Neoplasias Hepáticas/inducido químicamente , Animales , Masculino , Extractos Vegetales/análisis , Ratas , Ratas Endogámicas F344 , RubiaRESUMEN
Madder color (MC) has been shown to exert carcinogenic potential in the rat kidney in association with degeneration, karyomegaly, increased cell proliferation of renal tubule cells and increased renal 8-OHdG levels. To clarify the causal relationship of components and metabolites of MC to renal carcinogenesis, male F344 rats were fed lucidin-3-O-primeveroside (LuP) or alizarin (Alz), and the genotoxic LuP metabolites lucidin (Luc) or rubiadin (Rub) for up to 26 weeks. After one week and four weeks, Luc did not induce any renal changes. In contrast, after one week, cortical tubule degeneration was apparent in the Alz and LuP groups, and cytoplasmic swelling with basophilic change and karyomegaly in the outer medulla was observed only in the Rub group. LuP and Rub increased the proliferative activity of tubule cells in the outer medulla, and Alz and LuP increased renal 8-OHdG levels. After 26 weeks, Rub but not Alz induced atypical tubules, a putative preneoplastic lesion, and karyomegaly in the outer medulla. These results indicate that Rub may be a potent carcinogenic metabolite of MC, targeting proximal tubule cells in the outer medulla, although oxidative stress increased by Alz or LuP might also be involved in renal carcinogenesis by MC.
Asunto(s)
Antraquinonas/toxicidad , Neoplasias Renales/inducido químicamente , Extractos Vegetales/toxicidad , Rubia/toxicidad , 8-Hidroxi-2'-Desoxicoguanosina , Animales , Desoxiguanosina/análogos & derivados , Desoxiguanosina/análisis , Relación Dosis-Respuesta a Droga , Riñón/efectos de los fármacos , Riñón/patología , Masculino , Tamaño de los Órganos/efectos de los fármacos , Extractos Vegetales/metabolismo , Antígeno Nuclear de Célula en Proliferación/análisis , Ratas , Ratas Endogámicas F344 , Rubia/metabolismoRESUMEN
Madder color (MC) extracted from the roots of Rubia tinctorum (madder root) has been used as a food coloring in Japan. Our previous studies revealed MC to have obvious subchronic and chronic toxicity and potent carcinogenicity targeting rat liver and kidney. In the present two-year carcinogenicity study, conducted to further elucidate the long-term effects of MC and its target organs, male and female F344 rats were fed diet containing 0%, 2.5%, and 5.0% MC for 104 weeks. Body weights were significantly decreased in treated groups of both sexes throughout the feeding period. However, survival rates at week 104 were higher in treated groups of both sexes than in controls. Relative weights of the kidneys and liver were significantly increased in treated groups of both sexes. Histopathologically, karyomegaly and atypical tubules/hyperplasias, as well as renal cell adenomas and carcinomas were significantly increased in treated groups of both sexes with dose-dependence. Moreover, the incidence of hepatocellular adenomas and/or carcinomas was increased significantly with a dose-relation in treated groups of both sexes. These data provide clear evidence that MC exerts unequivocal carcinogenicity against renal tubule cells and hepatocytes in rats.
Asunto(s)
Aditivos Alimentarios/toxicidad , Neoplasias Renales/inducido químicamente , Neoplasias Hepáticas/inducido químicamente , Extractos Vegetales/toxicidad , Rubia/toxicidad , Adenoma/inducido químicamente , Adenoma/patología , Animales , Carcinoma/inducido químicamente , Carcinoma/patología , Relación Dosis-Respuesta a Droga , Esquema de Medicación , Femenino , Riñón/patología , Neoplasias Renales/patología , Hígado/patología , Neoplasias Hepáticas/patología , Ganglios Linfáticos/patología , Masculino , Ratas , Ratas Endogámicas F344 , Caracteres SexualesRESUMEN
To evaluate chronic toxicity of madder color (MC), a natural food colorant extracted from the roots of Rubia tinctorum L., F344 rats were fed diet containing 0%, 0.2%, 1.0% or 5.0% MC for 53 weeks. Hematological changes including anemia and serum biochemical alterations indicating hepatotoxicity were demonstrated at 5.0% in both sexes. Relative weights of the liver were significantly increased from 1.0% in both sexes, and those of the kidney were significantly increased from 1.0% in males and from 0.2% in females. Histopathologically, atypical renal tubule hyperplasias were increased at 1.0% or higher in both sexes in association with increase of cell proliferative activity in the tubules. A renal cell adenoma was observed in a male rat receiving 5.0% MC. In addition, glutathione S-transferase placental form-positive liver cell foci were significantly increased at 5.0% in both sexes. These results indicate that MC has chronic toxicity targeting kidney, liver and blood cells. Moreover, the results strongly suggest that MC may have the carcinogenic potential in the kidney and the liver.
Asunto(s)
Neoplasias Renales/inducido químicamente , Neoplasias Hepáticas/inducido químicamente , Extractos Vegetales/administración & dosificación , Extractos Vegetales/toxicidad , Lesiones Precancerosas/inducido químicamente , Rubia/toxicidad , Animales , Proliferación Celular , Relación Dosis-Respuesta a Droga , Esquema de Medicación , Femenino , Riñón/citología , Neoplasias Renales/patología , Neoplasias Hepáticas/patología , Masculino , Ratas , Ratas Endogámicas F344 , Pruebas de Toxicidad CrónicaRESUMEN
To identify genes linked to early stages of disruption of brain sexual differentiation, hypothalamic region-specific microarray analyses were performed using a microdissection technique with neonatal rats exposed to endocrine-acting drugs. To validate the methodology, the expression fidelity of microarrays was first examined with two-round amplified antisense RNAs (aRNAs) from methacarn-fixed paraffin-embedded tissue (PET) in comparison with expression in unfixed frozen tissue (UFT). Decline of expression fidelity when compared with the 1x-amplified aRNAs from UFTs was found as a result of the preferential amplification of the 3' side of mRNAs in the second round in vitro transcription. However, expression patterns for the 2x-amplified aRNAs were mostly identical between methacarn-fixed PET and UFT, suggesting no obvious influence of methacarn fixation and subsequent paraffin embedding on expression levels. Next, in the main experiment, neonatal rats at birth were treated subcutaneously either with estradiol benzoate (EB; 10 microg/pup) or flutamide (FA; 250 microg/pup), and medial preoptic area (MPOA)-specific microarray analysis was performed 24 h later using 2x-amplified aRNAs from methacarn-fixed PET. Numbers of genes showing constitutively high expression in the MPOA predominated in males, implying a link with male-type growth supported by perinatal testosterone. Around 60% of genes showing sex differences in expression demonstrated altered levels after EB treatment in females, suggesting an involvement of genes necessary for brain sexual differentiation. When compared with EB, FA affected a rather small number of genes, but fluctuation was mostly observed in females, as with EB. Moreover, many selected genes common to EB and FA showed down-regulation in females with both drugs, suggesting a common mechanism for endocrine center disruption in females, at least at early stages of post-natal development.
Asunto(s)
Antagonistas de Andrógenos/farmacología , Anticonceptivos/farmacología , Estradiol/análogos & derivados , Flutamida/farmacología , Expresión Génica/efectos de los fármacos , Hipotálamo/efectos de los fármacos , Diferenciación Sexual/efectos de los fármacos , Ácido Acético/farmacología , Animales , Cloroformo/farmacología , Estradiol/farmacología , Femenino , Fijadores , Secciones por Congelación , Perfilación de la Expresión Génica/métodos , Regulación del Desarrollo de la Expresión Génica , Hipotálamo/metabolismo , Masculino , Metanol/farmacología , Análisis por Micromatrices/métodos , Microdisección , RatasRESUMEN
Dietary fibers and chlorophyllin have shown to exert anti-carcinogenic effects against co-administered carcinogens. To test the possibility of chemoprevention by such dietary supplements on subacutely induced acrylamide (ACR) toxicity, Sprague-Dawley male rats were administered 2.5% sodium alginate, 5% glucomannan, 5% digestion resistant maltodextrin, 2.5% chitin or 1% chlorophyllin in the diet, and starting one week later, co-administered 0.02% ACR in the drinking water for 4 weeks. For comparison, untreated control animals given basal diet and tap water were also included. Neurotoxicity was examined with reference to gait abnormalities and by quantitative assessment of histopathological changes in the sciatic and trigeminal nerves, as well as aberrant dot-like immunoreactivity for synaptophysin in the cerebellar molecular layer. Testicular toxicity was assessed by quantitation of seminiferous tubules with exfoliation of germ cells into the lumen and cell debris in the ducts of the epididymides. Development of testicular toxicity as well as neurotoxicity was evident with ACR-treatment, but was not suppressed by dietary addition of fibers or chlorophyllin, suggesting no apparent beneficial influence of these dietary supplements on experimentally induced subacute ACR toxicity.
Asunto(s)
Acrilamida/toxicidad , Clorofilidas/farmacología , Fibras de la Dieta/farmacología , Enfermedades del Sistema Nervioso/inducido químicamente , Enfermedades del Sistema Nervioso/prevención & control , Alginatos/farmacología , Animales , Peso Corporal/efectos de los fármacos , Quitina/farmacología , Ingestión de Líquidos/efectos de los fármacos , Marcha/efectos de los fármacos , Ácido Glucurónico/farmacología , Ácidos Hexurónicos/farmacología , Histocitoquímica , Masculino , Mananos/farmacología , Enfermedades del Sistema Nervioso/patología , Tamaño de los Órganos/efectos de los fármacos , Polisacáridos/farmacología , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Testículo/patologíaRESUMEN
Jamaica quassia extract (JQE), a natural bittering agent, was investigated for hepatocarcinogenesis-promoting potential using a medium-term liver bioassay system. F344 male rats were given a single intraperitoneal injection of diethylnitrosamine (200mg/kg body weight) and then starting 2 weeks later, received JQE in the diet at concentrations of 500, 5000 or 30,000 ppm for 6 weeks. Animals for tumor promotion (+) and (-) controls were fed 500 ppm sodium phenobarbital (PB) and basal diet, respectively during the promotion phase in this model. All animals were subjected to two-thirds partial hepatectomy at week 3 and killed at week 8. As with the PB-promoted case, both numbers and areas of glutathione S-transferase placental form-positive liver cell foci were significantly increased by JQE at 30,000 ppm, with non-significant increases evident at 5000 ppm. The results thus indicate that JQE at high dose has promoting potential for rat hepatocarcinogenesis.
Asunto(s)
Carcinógenos/toxicidad , Neoplasias Hepáticas Experimentales/inducido químicamente , Picrasma/química , Extractos Vegetales/toxicidad , Lesiones Precancerosas/inducido químicamente , Quassia/química , Animales , Carcinógenos/administración & dosificación , Dieta , Dietilnitrosamina/administración & dosificación , Dietilnitrosamina/toxicidad , Glutatión Transferasa/metabolismo , Hepatectomía , Hepatocitos/efectos de los fármacos , Hepatocitos/enzimología , Hepatocitos/patología , Inyecciones Intraperitoneales , Hígado/efectos de los fármacos , Hígado/enzimología , Hígado/patología , Neoplasias Hepáticas Experimentales/enzimología , Neoplasias Hepáticas Experimentales/patología , Masculino , Extractos Vegetales/administración & dosificación , Lesiones Precancerosas/enzimología , Lesiones Precancerosas/patología , Ratas , Ratas Endogámicas F344RESUMEN
A subchronic toxicity study of water pepper extract (WPE) from Polygonum hydropiper L. was conducted in groups of 10 male and 10 female F344 rats fed powdered diets containing 0, 62.5, 250, 1000 or 4000 ppm concentrations for 13 weeks. Suppression of body weight gain due to decreased food consumption was observed in both sexes at 4000 ppm, and at autopsy, increase of relative weights was observed for the brain, liver, spleen, kidneys, and testes in these animals, suggestive of the reflection of the reduced body weights. At this dose, slight increases of blood urea nitrogen in both sexes and serum alanine aminotransferase, Na and Cl in females, were observed, suggestive of weak hepatic and renal toxicity, at least in females. The same females also exhibited slight decrease of red blood cells and haematocrit, slight increase of mean corpuscular volume and mean corpuscular haemoglobin, and minimal increase of splenic haemosiderin deposition, providing evidence of slight haemolytic anemia. On the other hand, enhanced accumulation of mast cells was observed in the mesenteric lymph nodes at 4000 ppm in males and 1000 and 4000 ppm in females. Considering the anti-anaphylactic properties of polygodial, a major constituent of WPE, the mast cell accumulation was concluded to be an adaptive change in response to the subchronic oral administration of WPE. Based on the present toxicity data, 1000 ppm was determined to be the no-observed-adverse-effect level, translating into 57.4 and 62.9 mg/kg/day for male and female rats, respectively.