RESUMO
Recently, cornifin-alpha/SPPR1 has been identified as a putative precursor protein of the cornified cell envelope. In this study, the expression and localization of cornifin-alpha/SPPR1 was examined in untreated and tumor promoter-treated mouse skin, hair follicles, and skin neoplasms. Western analysis with antiserum (SQ37A) to a rabbit cornifin-alpha peptide or antiserum (SQ37C) to a human SPRR1 peptide demonstrated a 31-kDa immunoreactive protein in mouse epidermis and Northern analysis revealed the presence of a 1-kb mRNA. Immunohistochemical staining of mouse skin with SQ37A or SQ37C revealed intense and specific staining of the infundibulum, isthmus, and of Henle's layer of the inner root sheath of the lower anagen hair follicle and weak staining of the telogen follicle and the suprabasal layers of the epidermis. Treatment of mouse skin with 12-0-tetradecanoyl-phorbol-13-acetate (TPA) produced a large increase in cornifin-alpha/SPRR1 protein and mRNA. Immunohistochemical localization of cornifin-alpha/SPRR1 in TPA-treated skin indicated that cornifin-alpha/SPRR1 was increased in the suprabasal epidermis but not in the follicle. sn-1,2,-didecanoylglycerol, a model lipid second messenger, produced an increase in cornifin-alpha/SPRR1 protein similar to that of TPA, while mirex, a non-phorbol ester-type promoter had no effect. Topical doses of retinoic acid did not repress TPA-induced cornifin-alpha/SPRR1 expression. Papillomas demonstrated a 10- and 100-fold increase in cornifin-alpha/SPRR1 protein and mRNA, and expression was restricted to suprabasal cells. Squamous cell carcinomas exhibited an intermediate level of cornifin-alpha protein, and expression was restricted to keratinized areas. These data indicate: i) cornifin-alpha/SPRR1 is expressed in mouse skin; ii) cornifin-alpha/SPRR1 is localized to specific areas of the anagen hair follicle with weak staining in the telogen follicle and epidermis; iii) epidermal cornifin-alpha/SPRR1 expression is induced by phorbol ester and sn-1,2-didecanoylglycerol but not mirex, and iv) papillomas and squamous cell carcinomas demonstrate a constitutive increase in cornifin-alpha/SPRR1 in differentiated areas of the neoplasms.
Assuntos
Epiderme/química , Folículo Piloso/química , Proteínas de Membrana/análise , Proteínas/análise , Neoplasias Cutâneas/química , Animais , Carcinoma de Células Escamosas/induzido quimicamente , Carcinoma de Células Escamosas/química , Proteínas Ricas em Prolina do Estrato Córneo , Feminino , Imuno-Histoquímica , Proteínas de Membrana/genética , Camundongos , Mirex/farmacologia , Papiloma/induzido quimicamente , Papiloma/química , Proteínas/genética , RNA Mensageiro/análise , Acetato de Tetradecanoilforbol/farmacologiaRESUMO
The thyroid glands, from rats which had received photomirex (0.05, 0.5, 5 or 50 ppm) or mirex (5 or 50 ppm) in their diets for 28 days and maintained for 18 months on clean diet, were examined by electron microscopy. A dose of 0.05-5 ppm photomirex resulted in an augmentation of principal follicular cell heights. Their cytoplasm exhibited a marked numerical increase of secondary lysosomes. In addition, at 5 ppm, follicular cells in some segments of the gland contained only a few secondary lysosomes. After dietary exposure to 50 ppm photomirex the number of lysosomal elements remained elevated in epithelial cell cytoplasm. Morphological aberrations in the follicles could not be detected after the 5 ppm mirex treatment. The most remarkable alterations in this experiment occurred in the thyroid glands of animals ingesting 50 ppm mirex where columnar follicular cells were engorged with deformed lysosomal bodies. Remaining components of the follicular cell architecture in all the treated groups were unaffected and were similar to those in the control group. These results demonstrated that following a 28-day dietary exposure of rats to photomirex (0.05-50 ppm) or mirex (50 ppm), alterations in the thyroid glands persisted for at least 18 months.
Assuntos
Inseticidas/farmacologia , Mirex/farmacologia , Glândula Tireoide/efeitos dos fármacos , Administração Oral , Animais , Masculino , Microscopia Eletrônica , Mirex/análogos & derivados , Ratos , Ratos Endogâmicos , Glândula Tireoide/ultraestrutura , Fatores de TempoRESUMO
The effect of prior hypophysectomy upon mirex-induced liver hypertrophy in male Sprague-Dawley rats was examined. Mirex had no effect upon adrenal weight, liver weight, plasma glucose or plasma corticosterone in hypophysectomized rats. However, daily corticosterone supplements (20 mg/kg body weight, sc) given to mirex-treated hypophysectomized animals yielded a 52% increase in liver weight to body weight ratios over those observed in mirex-treated hypophysectomized animals not receiving supplement. In intact rats, both liver weight to body weight ratios and plasma ACTH were significantly increased over controls 2 days after mirex treatment. These results indicate that mirex-induced liver enlargement not only requires corticosterone, but that the response is dependent upon an intact pituitary-adrenalcortical axis.
Assuntos
Córtex Suprarrenal/fisiologia , Inseticidas/farmacologia , Fígado/anatomia & histologia , Mirex/farmacologia , Hipófise/fisiologia , Glândulas Suprarrenais/anatomia & histologia , Hormônio Adrenocorticotrópico/sangue , Animais , Corticosterona/sangue , Corticosterona/farmacologia , Hipofisectomia , Masculino , Tamanho do Órgão/efeitos dos fármacos , Ratos , Ratos EndogâmicosRESUMO
Previous work has established the marked potentiation of CCl4 hepatoxicity by prior exposure to chlordecone (CD). This study was conducted to determine if prior exposure to CD results in enhancement of CCl4-induced destruction of the hepatic microsomal mixed-function oxygenase (MFO) system. Male Sprague-Dawley rats received a single oral dose of CD (10 mg/kg) or corn oil vehicle alone (1 ml/kg) 24 hr prior to a single ip injection of CCl4 (0-100 microliter/kg). Mirex (M; 10 mg/kg) and phenobarbital (PB; 80 mg/kg/day for two days) were used as negative and positive controls respectively for the potentiation of CCl4 hepatotoxicity. Hepatotoxicity was evaluated 24 hrs after CCl4 administration by elevations of three serum enzymes (GPT, GOT, and ICD). The key hepatic microsomal MFO parameters measured were microsomal protein, cytochrome P-450 content, glucose-6-phosphatase (G-6-Pase), and aminopyrine demethylase (APD). As previously demonstrated using a subchronic dietary pretreatment protocol, CD potentiated CCl4 hepatotoxicity over a range of CCl4 doses to a greater extent than PB or M, as judged by elevations in serum enzymes. PB caused the greatest increase in total P-450 content and the greatest increase in CCl4-mediated destruction of microsomal protein and APD activity. M caused the least destruction of total hepatic cytochrome P-450, despite the same level of cytochrome P-450 as in the PB group. CD treatment caused the greatest decrease in G-6-Pase activity in comparison to PB or M pretreatments and a similar degree of P-450 destruction as observed with the PB group. These findings suggest that in general, CCl4-induced destruction of hepatic MFO parameters measured in this study is disproportional to the known degree of potentiated hepatotoxicity by the pretreatments and does not accurately reflect the potentiation of CCl4 hepatotoxicity by CD.
Assuntos
Tetracloreto de Carbono/farmacologia , Clordecona/farmacologia , Inseticidas/farmacologia , Mirex/farmacologia , Oxigenases de Função Mista/antagonistas & inibidores , Fenobarbital/farmacologia , Alanina Transaminase/sangue , Aminopirina N-Desmetilase/antagonistas & inibidores , Animais , Aspartato Aminotransferases/sangue , Sistema Enzimático do Citocromo P-450/metabolismo , Sinergismo Farmacológico , Glucose-6-Fosfatase/antagonistas & inibidores , Isocitrato Desidrogenase/sangue , Masculino , Microssomos Hepáticos/efeitos dos fármacos , Microssomos Hepáticos/enzimologia , Ratos , Ratos EndogâmicosRESUMO
The hepatic induction of cytochrome P-450-dependent monooxygenase components by cis- and trans-5,10-dihydrogen mirex was studied in male and female laboratory rats. There were sex-dependent differences between the two isomeric derivatives of mirex. The cis-isomer significantly increased aniline hydroxylase activity in the female, but not in the male. In contrast, aminopyrine N-demethylase was significantly increased by the cis-isomer in both sexes. The trans-isomer increased the hydroxylase and N-demethylase activities in both sexes. The cis-isomer induced NADPH-cytochrome c reductase in the female, and the trans-isomer did not. Both isomers induced hepatic reductase activity in the male rat. No sex-dependent differences in the hepatic induction of cytochrome P-450 were observed for either isomer.
Assuntos
Inseticidas/farmacologia , Fígado/enzimologia , Mirex/farmacologia , Oxigenases/biossíntese , Aminopirina N-Desmetilase/biossíntese , Anilina Hidroxilase/biossíntese , Animais , Sistema Enzimático do Citocromo P-450 , Indução Enzimática , Feminino , Masculino , Mirex/análogos & derivados , Mirex/metabolismo , Ratos , Ratos Endogâmicos , Fatores SexuaisRESUMO
The effects of chlordecone (CD), mirex or phenobarbital (PB) treatment of male Mongolian gerbils on CCl4-hepatotoxicity were determined. These monooxygenase inducers did not potentiate CCl4-hepatotoxicity in the gerbil although phenobarbital and CD are known potentiators in the rat. The control gerbil was more sensitive to CCl4-hepotoxicity than the control rat as reflected in loss of cytochrome (cyt) P-450 and increases in SGOT and SGPT. In addition, CCl4 treatment of the control gerbils resulted in significant reductions in cyt b5 and NADPH-cyt P-450 levels, a phenomenon not observed in the rat.
Assuntos
Intoxicação por Tetracloreto de Carbono/enzimologia , Fígado/enzimologia , Oxigenases de Função Mista/biossíntese , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Clordecona/farmacologia , Grupo dos Citocromos b/metabolismo , Citocromos b5 , Indução Enzimática , Gerbillinae , Masculino , Mirex/farmacologia , NADPH-Ferri-Hemoproteína Redutase/metabolismo , Fenobarbital/farmacologiaRESUMO
Male Sprague-Dawley rats (200-250 g) were maintained on a normal powdered diet or on a similar diet containing 10 ppm chlordecone (CD), 10 ppm mirex (M) or 225 ppm phenobarbital (PB) for 15 days. On day 15, the animals received a single i.p. administration of CCl4 (100 microliters/kg). Hepatic DNA, RNA, protein, lipid and glycogen were determined 1, 4, 6, 12, 24 and 36 h after CCl4 administration. A significant decrease (18%) in hepatic protein was observed 24 h after CCl4 challenge in the CD-pretreated rats; significant changes were not observed in the other treatment groups. Hepatic RNA was decreased (37%) in CD-pretreated rats at 36 h; no changes were observed in the DNA content. Hepatic RNA and DNA were increased (20% and 16%, respectively) 6 h after exposure to CCl4 alone. Lipid content was increased at all time points starting at 4 h in response to CCl4 challenge in the CD-pretreated rats. In the M- and PB-pretreated rats increases in hepatic lipid (22 and 28%, respectively) were observed only at the 6-h time point. Only a transient increase occurred after CCl4 alone at 4 h. While depletion of hepatic glycogen was manifested in all groups at all time points following CCl4, that in the CD + CCl4 group was the greatest. A recovery of glycogen beginning at 12 h was observed in the rats receiving CCl4 alone and in the M and PB pretreated animals. No such recovery was evident in CD + CCl4 group. These studies indicate that biochemical changes compatible with cellular death are more pronounced in the CD-pretreated rats than in those receiving CCl4 alone, suggesting that the metabolic and supportive biochemical mechanisms for hepatocellular repair are suppressed in rats receiving CD + CCl4.
Assuntos
Tetracloreto de Carbono/farmacologia , Clordecona/farmacologia , Inseticidas/farmacologia , Fígado/efeitos dos fármacos , Mirex/farmacologia , Fenobarbital/farmacologia , Animais , DNA/análise , Lipídeos/análise , Fígado/análise , Glicogênio Hepático/análise , Masculino , Proteínas/análise , RNA/análise , Ratos , Ratos EndogâmicosRESUMO
The effects of mirex, two monohydrogen and two dihydrogen mirex derivatives, and chlordecone on several hepatic parameters were studied 2 days following a single oral dose of 100 mg/kg in female rats. All compounds increased microsomal cytochrome P-450 content, NADPH-cytochrome c reductase activity, and hepatic ascorbic acid concentration. Microsomal protein concentration was generally increased. All compounds except chlordecone increased relative liver weight and the activities of aminopyrine N-demethylase and p-nitroanisole O-demethylase. Hepatic concentration of protein and glutathione were unaltered. The dechlorinated mirex derivatives caused effects of a magnitude similar to that of mirex, whereas chlordecone was considerably less potent.
Assuntos
Clordecona/farmacologia , Inseticidas/farmacologia , Microssomos Hepáticos/efeitos dos fármacos , Mirex/farmacologia , Oxigenases de Função Mista/metabolismo , Oxirredutases/metabolismo , Animais , Sistema Enzimático do Citocromo P-450/metabolismo , Feminino , Hepatomegalia/induzido quimicamente , Microssomos Hepáticos/enzimologia , Mirex/análogos & derivados , Ratos , Ratos EndogâmicosRESUMO
Interactions of chlordecone (kepone) and mirex with nicotinic acetylcholine (ACh) receptor complexes from Torpedo californica electric organs were studied using biochemical probes for ACh and ion-channel binding sites. Neither compound inhibited the binding of [125I] alpha-bungarotoxin (BGT) to the receptor; chlordecone, however, enhanced carbamylcholine affinity for the receptor 5-fold. Chlordecone, but not mirex, also inhibited the binding of [3H]perhydrohistrionicotoxin and [3H]phencyclidine to sites associated with the receptor-gated ion channel. Ion-channel inhibition by chlordecone was enhanced in the presence of carbamylcholine. These results indicate that chlordecone, but not mirex, interacts with the ion-translocation mechanism associated with nicotinic ACh receptors, where it may sterically block ion flux as well as stabilize a desensitized conformation of the receptor complex.
Assuntos
Clordecona/farmacologia , Inseticidas/farmacologia , Canais Iônicos/efeitos dos fármacos , Mirex/farmacologia , Receptores Nicotínicos/efeitos dos fármacos , Regulação Alostérica/efeitos dos fármacos , Venenos de Anfíbios/metabolismo , Animais , Bungarotoxinas/metabolismo , Carbacol/farmacologia , Órgão Elétrico/metabolismo , Técnicas In Vitro , Canais Iônicos/metabolismo , Fenciclidina/metabolismo , Receptores Nicotínicos/metabolismo , TorpedoRESUMO
A polychlorinated biphenyl mixture, Aroclor 1254, two commercial grade insecticides, mirex and kepone, and a pesticide breakdown product, 1,2,4-trichlorobenzene were evaluated for mutagenicity and hepatic enzyme induction potential in the Salmonella/microsomal assay. None was found to revert strains TA1535, TA1537, TA98 or TA100 when tested with or without metabolic activation. Liver microsomal extracts (S9) from rats induced with 1,2,4-trichlorobenzene were shown to differ from S9 of either control or Aroclor 1254-induced rats in the capacity to activate 2-aminoanthracene mutagenesis.
Assuntos
Arocloros/farmacologia , Clordecona/farmacologia , Clorobenzenos/farmacologia , Inseticidas/farmacologia , Mirex/farmacologia , Mutagênicos , Bifenilos Policlorados/farmacologia , Avaliação Pré-Clínica de Medicamentos , Técnicas Genéticas , Salmonella typhimurium/genéticaRESUMO
Chlordecone is known to bring about alterations in the endocrine systems of a variety of animals. The present study was undertaken to investigate the possibility of a direct effect of the insecticide on steroid-secreting tissues. Technical grade chlordecone produced a dose-dependent inhibition of steroidogenesis in cultured mouse adrenal tumor cells (Y-1 cells) when stimulated with ACTH (IC50 = 4 X 10(-5) M), cAMP (IC50 = 2.3 X 10(-5) M), or pregnenolone (IC50 3.5 X 10(-5) M). Similar values were obtained with 99% pure chlordecone. These data suggest that chlordecone inhibited the conversion of pregnenolone into 20 alpha-dihydroprogesterone. Additional sites of action cannot be ruled out. Mirex failed to inhibit steroidogenesis which was supported by exogenous pregnenolone. At a concentration of 3 X 10(-6) M mirex inhibited cAMP induced steroidogenesis, but higher concentrations did not produce a more pronounced effect.