RESUMEN
There is compelling evidence that a hypercaloric, high-fructose diet can cause metabolic syndrome (MetS) and a whole range of other metabolic changes. In the context of androgen deficiency, MetS in boys merits special attention, but the effects of fructose-rich diet in youth on future male reproductive function are still poorly evidenced. The aim of this study was to address this issue and analyse the effects of high-fructose intake starting from weaning to puberty (postnatal day 23 up to 83) on the reproductive function of male rats. For this purpose juvenile male Wistar rats were divided in two groups: control and the group receiving 10 % fructose solution instead of drinking water. Reproductive function was evaluated in terms of fertility, sperm count, testes/epididymis morphology, and serum sex hormones. The fructose-treated group showed a decrease in testosterone and twofold increase in luteinising and follicle-stimulating hormone levels in the serum. This was accompanied with lower testis/epididymis weights, sperm count, and changed testis/epididymis morphology. Their fertility remained unchanged, but the fertility of females mating with these males diminished. In addition, pre-implantation and post-implantation embryonic death rate rose in these females. Our results have confirmed that high fructose consumption from early age until puberty can impair the reproductive function of male rats, and call for further animal and epidemiological investigation.
Asunto(s)
Fertilidad/efectos de los fármacos , Jarabe de Maíz Alto en Fructosa/efectos adversos , Reproducción/efectos de los fármacos , Maduración Sexual/efectos de los fármacos , Recuento de Espermatozoides , Espermatogénesis/efectos de los fármacos , Testosterona/sangre , Animales , Masculino , Ratas , Ratas WistarRESUMEN
Despite clear beneficial effects, studies on the efficacy of different formulations of micronutrients and vitamins in the treatment of male infertility are still very limited. The aim of the study was to compare the efficacy of methionine and novel formulation Metovitan based on the combination of methionine, thiamine, nicotinamide, α-tocopherol acetate and zinc salt as remedies for prevention of anti-tuberculosis drugs (ATD) anti-fertility effects in male rats. ATD co-administration resulted in testicular CYP2E1, CYP2C23 and CYP3A2 transcriptional activation. Methionine and Metovitan regulated the level of CYP2E1 and CYP3A2 mRNA expression. Methionine unaffected CYP2C23 mRNA level, while Metovitan effectively normalized this parameter. The use of methionine did not allow normalizing of DNA fragmentation processes in testes, while Metovitan provided decrease in the number of DNA fragments to the control level. Both substances significantly decreased p-nitrophenol hydroxylase activity and prevented pro/antioxidant parameters imbalance in testes of ATD-treated rats. The end results of methionine and Metovitan application were partial or complete restoration of altered spermatogenesis parameters with subsequent increase in sperm count and fertility. Ameliorating effects may be attributed to antioxidant properties and modulating effects on testicular CYPs. More pronounced beneficial Metovitan effects might be due to synergic effects of its components.
Asunto(s)
Antituberculosos/toxicidad , Epidídimo/efectos de los fármacos , Infertilidad Masculina/tratamiento farmacológico , Metionina/química , Metionina/farmacología , Testículo/efectos de los fármacos , Animales , Citocromo P-450 CYP2J2 , Sistema Enzimático del Citocromo P-450/efectos de los fármacos , Combinación de Medicamentos , Epidídimo/patología , Infertilidad Masculina/inducido químicamente , Masculino , Ratas , Ratas Wistar , Espermatogénesis/efectos de los fármacos , Testículo/patologíaRESUMEN
BACKGROUND: Complex investigations of cytochrome P450 (CYP) isoforms with metabolic syndrome (MS) development are limited, and specific features of adolescent's metabolisms are generally disregarded. The aim of present study was a comparative estimation of MS-mediated changes in CYP3A, CYP2C, and CYP2E1 mRNA expression and enzymatic activities, as well as antioxidant system parameters of adult and pubertal rats. METHODS: Wistar albino male rats of two age categories [young animals of 21 days age (50-70 g) and adults (160-180 g)] were divided into four groups (eight animals in each group): (1) control 1 (intact young rats), (2) control 2 (intact adult rats), (3) MS3 (young rats with MS), and (4) MS4 (adult rats with MS). The MS was induced by full replacement of drinking water by 20% fructose solution (200 g/L). After 60 days of MS modeling, the investigation of rat liver CYP3A, CYP2C, and CYP2E1 mRNA expressions, their enzyme-marker activities, as well as the antioxidant system parameters was conducted. RESULTS: Levels of liver CYP2E1 mRNA expression increased with MS: 40% (adults) and 80% (pubertal rats). Pubertal rats had also increased CYP3A2 mRNA expression (30%) and decreased CYP2C mRNA expression (30%). Changes in CYP2E1 and CYP2C enzymatic activities were consistent with the changes of corresponding gene expressions at both age-groups with MS. Simultaneously, liver reduced glutathione contents, and glutathione transferase and reductase activities were decreased in pubertal animals. CONCLUSIONS: CYP isoform expression rates and glutathione system were greatly violated with MS. The greater changes were observed in pubertal rats with MS.
Asunto(s)
Citocromo P-450 CYP2E1/biosíntesis , Citocromo P-450 CYP3A/biosíntesis , Sistema Enzimático del Citocromo P-450/biosíntesis , Síndrome Metabólico/enzimología , Maduración Sexual/fisiología , Factores de Edad , Animales , Citocromo P-450 CYP2E1/farmacología , Isoenzimas/biosíntesis , Hígado/enzimología , Masculino , Ratas , Ratas WistarRESUMEN
Isoniazid is reported to be the most reliable and cost-effective remedy for tuberculosis treatment and prophylaxis among first line anti-tuberculosis drugs. Conventionally, the most common and best studied adverse effect of isoniazid is hepatotoxicity, but as for testicular toxicity the problem has not yet explored extensively. The aim of the study was to identify in vivo influence of isoniazid on induction of testicular cytochrome Ð -450 2Ð1 (CYP2E1) mRNA expression and enzymatic activity, testes DNA fragmentation, serum total testosterone level, and spermatogenesis indices. The significant induction of CYP2E1 was demonstrated in rat's testes following isoniazid administration, specifically CYP2E1 mRNA expression and p-nitrophenolhydroxylase activity was increased in 28 and 7 times as compared with control, respectively. These changes were accompanied by activating of testicular GST in 32%, changing in levels and character of DNA fragmentation, as well as damaging of the spermatogenic epithelium, decreasing in serum testosterone content (1.62 fold), sperm count (19%), and losing of fertility in comparison with untreated males. We assume that in testes of isoniazid-treated rats CYP2E1 may act as a trigger in generating of reactive oxygen species and other toxic metabolites which subsequently mediates DNA damage, spermatogenesis disturbances, and altered male fertilizing capacity.
Asunto(s)
Antituberculosos/toxicidad , Inductores del Citocromo P-450 CYP2E1/toxicidad , Citocromo P-450 CYP2E1/biosíntesis , Isoniazida/toxicidad , Espermatozoides/efectos de los fármacos , Enfermedades Testiculares/inducido químicamente , Testículo/efectos de los fármacos , Animales , Apoptosis/efectos de los fármacos , Biomarcadores/sangre , Citocromo P-450 CYP2E1/genética , Fragmentación del ADN/efectos de los fármacos , Inducción Enzimática , Fertilidad/efectos de los fármacos , Infertilidad Masculina/inducido químicamente , Infertilidad Masculina/fisiopatología , Masculino , Estrés Oxidativo/efectos de los fármacos , ARN Mensajero/biosíntesis , Ratas Wistar , Especies Reactivas de Oxígeno/metabolismo , Recuento de Espermatozoides , Espermatogénesis/efectos de los fármacos , Espermatozoides/enzimología , Espermatozoides/patología , Enfermedades Testiculares/sangre , Enfermedades Testiculares/enzimología , Enfermedades Testiculares/patología , Testículo/enzimología , Testículo/patología , Testículo/fisiopatología , Testosterona/sangre , Factores de TiempoRESUMEN
BACKGROUND: Despite of the wide spectrum of alcoholism experimental models, the majority of them are very specialized on the short list of investigated parameters and could not provide reproduction of complex metabolic changes in the rats. The aim of the present study was to estimate whether rats selected by high alcohol preference, allowed free access to 15% alcohol for 150 days, develop simultaneous multilevel disturbances of cell macromolecules structure, metabolism and oxidative/nitrosative stress. METHODS: Wistar albino male rats were divided into groups: I - rats selected by preferences to alcohol were used for chronic alcoholism modeling by replacing water with 15% ethanol (150 days), II - control. Contents of amino acids in serum, liver mRNA CYP2E1 and CYP3A2 expression, DNA fragmentation and lipid peroxidation levels, the reduced glutathione content, superoxide dismutase, catalase, iNOS and cNOS activities were evaluated. RESULTS: In serum of ethanol-treated rats contents of aspartic acid, serine, glycine, alanine and valine were decreased whereas contents of histidine, methionine and phenylalanine were increased. Liver CYP2E1, CYP3A2 mRNA expression, DNA fragmentation levels significantly elevated. Level of cNOS in ethanol-treated rat's hepatocytes was within the normal limits, whereas iNOS activity was raised 1.6 times. Liver pro- and anti-oxidant system alterations were shown. CONCLUSIONS: Rats' chronic 15% alcohol consumption (150 days) led solely to complex metabolomic changes at different levels, which simultaneously characterized cell macromolecules structure, metabolism, and oxidative/nitrosative stress. Rodent model of chronic alcoholism in the proposed modification could be an adequate and reasonably priced tool for further preclinical development and testing of pharmacotherapeutic agents.
Asunto(s)
Alcoholismo/fisiopatología , Fragmentación del ADN/efectos de los fármacos , Peroxidación de Lípido/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Consumo de Bebidas Alcohólicas/efectos adversos , Animales , Citocromo P-450 CYP2E1/genética , Citocromo P-450 CYP3A/genética , Modelos Animales de Enfermedad , Hígado/efectos de los fármacos , Hígado/patología , Masculino , ARN Mensajero/metabolismo , Ratas , Ratas WistarRESUMEN
PURPOSE: The aim of the study was to investigate the correlation between spermatogenesis disorders and CYP2E1 mRNA contents in testes of rats with experimental alcoholism or type I diabetes. MATERIAL/METHODS: Two pathological states characterized by CYP2E1 induction were simulated on Wistar male rats: experimental alcoholism and type I diabetes. As controls for each state, equal number of animals (of the same age and weight) were used. Morphological evaluation of rat testes was carried out. The spermatogenic epithelium state was estimated by four points system. CYP2E1 mRNA expression was rated by method of reverse transcriptase polymerase chain reaction. Pearson correlation coefficients were used for describing relationships between variables. RESULTS: The presence of alcoholism and diabetes-mediated quantitative and qualitative changes in male rat spermatogenic epithelium in comparison with norm has been demonstrated. The increased levels of testes CYP2E1 have been fixed simultaneously. CYP2E1 mRNA content negatively strongly correlated with spermatogenic index value (r=-0.99; P<0.001) and positively strongly correlated with epithelium desquamation occurrence (r=0.99; P<0.001) in testes of rats with chronic alcoholism. The strong correlation between CYP2E1 mRNA content and number of spermatogonia (r=0.99; P<0.001) and "windows" occurrence (r=0.96; P<0.001) has been fixed in diabetic rats testes. CONCLUSIONS: Present investigation has demonstrated that the testicular failure following chronic ethanol consumption and diabetes type I in male rats accompanied CYP2E1 mRNA over-expression in testes. The correlation between the levels of CYP2E1 mRNA in testes and spermatogenesis disorders allow supposing the involvement of CYP2E1 into the non-specific pathogenetic mechanisms of male infertility under above-mentioned pathologies.
Asunto(s)
Alcoholismo/fisiopatología , Citocromo P-450 CYP2E1/metabolismo , Diabetes Mellitus Tipo 1/complicaciones , Modelos Animales de Enfermedad , Inducción Enzimática , Infertilidad Masculina/etiología , Testículo/metabolismo , Animales , Citocromo P-450 CYP2E1/genética , Infertilidad Masculina/complicaciones , Infertilidad Masculina/metabolismo , Infertilidad Masculina/patología , Masculino , ARN Mensajero/metabolismo , Ratas Wistar , Espermatogénesis , Testículo/patologíaRESUMEN
There is good evidence for impairment of spermatogenesis and reductions in sperm counts and testosterone levels in chronic alcoholics. The mechanisms for these effects have not yet been studied in detail. The consequences of chronic alcohol consumption on the structure and/or metabolism of testis cell macromolecules require to be intensively investigated. The present work reports the effects of chronic alcoholism on contents of free amino acids, levels of cytochrome P450 3A2 (CYP3A2) mRNA expression and DNA fragmentation, as well as on contents of different cholesterol fractions and protein thiol groups in rat testes. Wistar albino male rats were divided into two groups: I - control (intact animals), II - chronic alcoholism (15% ethanol self-administration during 150 days). Following 150 days of alcohol consumption, testicular free amino acid content was found to be significantly changed as compared with control. The most profound changes were registered for contents of lysine (-53%) and methionine (+133%). The intensity of DNA fragmentation in alcohol-treated rat testes was considerably increased, on the contrary CYP3A2 mRNA expression in testis cells was inhibited, testicular contents of total and etherified cholesterol increased by 25% and 45% respectively, and protein SH-groups decreased by 13%. Multidirectional changes of the activities of testicular dehydrogenases were detected. We thus obtained complex assessment of chronic alcoholism effects in male gonads, affecting especially amino acid, protein, ATP and NADPH metabolism. Our results demonstrated profound changes in testes on the level of proteome and genome. We suggest that the revealed metabolic disorders can have negative implication on cellular regulation of spermatogenesis under long-term ethanol exposure.
RESUMEN
This study is a complex investigation of alcohol-mediated changes in CYP2E1 mRNA and protein expression in the testes, as well as spermatogenesis indices and type I collagen amino acid contents, in male rats. Wistar albino male rats were divided into two groups: I--control (intact animals), II--experimental (chronic alcoholism, exposure to a 15% ethanol aqueous solution during 150 days). The destructive changes in the spermatogenic epithelium were accompanied by a decrease in sperm number and motility time. CYP2E1 mRNA and protein expression were elevated in the testes 3 and 1.4 times, respectively. Also, significantly lower contents of lysine, glutamic acid, serine, proline, alanine, valine, and phenylalanine residues accompanied by an increase of hydroxyproline, glycine, and threonine residue contents were detected in the skin type I collagen of the experimental group. Chronic ethanol consumption caused testicular failure along with an overexpression of CYP2E1 mRNA and protein in the testes as well as quantitative changes in type I collagen amino acid contents. The profound alcohol-mediated changes in collagen type I amino acid contents may have affected the spermatogenic epithelium state. The modulation of testicular cytochrome P450 2E1 mRNA and protein expression could change the functioning of this isozyme in target organs and take part in the mechanism of ethanol gonadotoxicity.
Asunto(s)
Citocromo P-450 CYP2E1/genética , Citocromo P-450 CYP2E1/metabolismo , Etanol/toxicidad , ARN Mensajero/metabolismo , Espermatogénesis/efectos de los fármacos , Testículo/metabolismo , Consumo de Bebidas Alcohólicas/efectos adversos , Aminoácidos/análisis , Animales , Colágeno Tipo I/análisis , Masculino , Ratas , Ratas Wistar , Recuento de Espermatozoides , Motilidad Espermática/efectos de los fármacos , Testículo/químicaRESUMEN
Current therapeutic regimens with first-line antitubercular agents are associated with a high rate of adverse effects which can lead to therapeutic failure. Understanding the nature and the severity of these effects is important for treatment optimization. The aim of present study was to investigate pyrazinamide potential effects on male rats DNA fragmentation, amino acid composition of bone type I collagen, reproductive capability and their posterity antenatal and postnatal development. Wistar albino male rats (160-200 g b.w.) were divided into three groups: I--received pyrazinamide per os at a dose of 1000 mg/kg b.w./day, II--at a dose of 2000 mg/kg b.w./day, in both groups it was given for 60 days; III--control. After 60 days of the experiment, rats of the experimental (groups I and II) and control groups were mated with intact virgin females. The amino acids contents of male rat bone type I collagens were determined using amino acid analyzer, epididymis and testis DNA fragmentation--electrophoretically; posterity antenatal development indices and postnatal development--by standard procedures. The study of pyrazinamide effects (administered in different doses) on males bone type I collagen amino acid contents and testis DNA fragmentation demonstrated the presence of dose-dependent pyrazinamide-mediated quantitative and qualitative changes in male rat reproductive organs DNA and extracellular matrix proteins in comparison with control. Changes in nucleic acids and proteins structure were accompanied by alterations in processes of fertilization (with intact females), embryogenesis and by lowering of posterity survival.
Asunto(s)
Aminoácidos/metabolismo , Huesos/efectos de los fármacos , Colágeno Tipo I/metabolismo , Fragmentación del ADN/efectos de los fármacos , Efectos Tardíos de la Exposición Prenatal/inducido químicamente , Pirazinamida/toxicidad , Reproducción/efectos de los fármacos , Animales , Huesos/metabolismo , Femenino , Masculino , Embarazo , Ratas , Ratas WistarRESUMEN
Introduction. Current therapeutic regimens with first-line antitubercular agents are associated to a high rate of adverse effects which could cause pronounced changes in collagen's contents and structure. Investigation of these changes is very important for optimization of antitubercular therapy and minimization of treatment-caused harm. The aim of present paper was to investigate potential effect of pyrazinamide on male rats' cartilage type II collagen amino acid composition. Materials and Methods. Wistar albino male rats (160-200 g b.w.) were divided into three groups: I-received pyrazinamide per os at a dose of 1000 mg/kg b.w./day; II-at a dose of 2000 mg/kg b.w./day, in both groups it was given for 60 days; III-control. After 60 days of the experiment, rats of the experimental (groups I and II) and control groups were sacrificed and the amino acids contents of male rat cartilage type II collagens were determined using amino acid analyzer. Results and Discussion. The study of pyrazinamide effects (administered in different doses) on rat cartilage type II collagen amino acid contents demonstrated presence of dose-dependent pyrazinamide-mediated quantitative and qualitative changes in these rat extracellular matrix proteins in comparison with control.
RESUMEN
Necessity of tuberculosis chemotherapy adverse effects minimization requires a comprehensive evaluation of the effects of antitubercular drugs on reproductive system and extracellular matrix proteins. Wistar albino male rats (160-200 g) were divided into three groups: I--received pyrazinamide per os at a dose of 1000 mg/kg bw/day, II--at a dose of 2000 mg/kg bw/day, in both group it was given for 60 days; III--intact animals. The contents of amino acids in rat type I collagens were determined using an amino acid analyzer. Morphological analyses were carried out by an optical microscope. The study of the effects of pyrazinamide administered in different doses on type I collagen amino acid contents, testis cells morphologic and morphometric parameters and spermatogenesis demonstrated the presence of pyrazinamide-mediated quantitative and qualitative changes in male rat reproductive organs, spermatogenic epithelial cells and extracellular matrix proteins in comparison with norm.The largest number of changes were established at a dose 2000 mg/kg b.w./day. The observed collagen molecules changes could hence affect the properties and correct functioning of spermatogenic epithelium and other tissues of reproductive organs. They might be caused by pyrazinamide via cytochrome P450 2E1 induction, reactive oxygen species production or direct action of this compound on protein biosynthesis processes.