RESUMO
Background: The objective of the current study was comparing the impact of two staining techniques on semen morphological parameters and their influence on patient diagnosis. The ideal staining method should preserve cell integrity while providing detailed information. Methods: Semen samples from fifty men were stained using Diff-Quick or Spermac methods. Morphological parameters were classified based on the Tygerberg criteria, and final diagnosis was according to WHO manual guidelines. Statistical analysis was performed through conducting paired t-tests or Wilcoxon rank-sum tests, with GLIMMIX and Fisher's exact test for determining the significance (p≤0.05). Results: Both staining methods highlighted head and tail regions, with Spermac offering better visualization of the midpiece. Spermac demonstrated fewer normal spermatozoa (2.8±0.3%) compared to Diff-Quick (3.98±0.4%; p=0.0385). Midpiece abnormalities were more evident with Spermac (55.7±2.1%) than Diff-Quick (24.8±2.0%; p<0.0001). No significant difference was found in head and tail abnormalities (p>0.05). Conclusion: Diff-Quick staining resulted in a higher proportion of normal spermatozoa, primarily due to its midpiece evaluation. The choice of staining method significantly impacts the diagnosis of infertile males. These findings have important implications for clinical practice and future research, suggesting the need for further investigations to assess different staining methods and determine optimal diagnostic thresholds.
RESUMO
A new library of hybrid compounds that combine the functional parts of glibenclamide and pioglitazone was designed and developed. Compounds were screened for their antihyperglycemic effects on the glucose tolerance curve. This approach provided a single molecule that optimizes the pharmacological activities of two drugs used for the treatment of diabetes mellitus type 2 (DM2) and that have distinct biological activities, potentially minimizing the adverse effects of the original drugs. From a total of 15 compounds, 7 were evaluated in vivo; the compound 2; 4- [2- (2-phenyl-4-oxo-1,3-thiazolidin-3-yl) ethyl] benzene-1-sulfonamide (PTEBS) was selected to study its mechanism of action on glucose and lipid homeostasis in acute and chronic animal models related to DM2. PTEBS reduced glycemia and increased serum insulin in hyperglycemic rats, and elevated in vitro insulin production from isolated pancreatic islets. This compound increased the glycogen content in hepatic and muscular tissue. Moreover, PTEBS stimulated the uptake of glucose in soleus muscle through a signaling pathway similar to that of insulin, stimulating translocation and protein synthesis of glucose transporter 4 (GLUT4). PTEBS was effective in increasing insulin sensitivity in resistance rats by stimulating increased muscle glucose uptake, among other mechanisms. In addition, this compound reduced total triglycerides in a tolerance test to lipids and reduced advanced glycation end products (AGES), without altering lactate dehydrogenase (LDH) activity. Thus, we suggest that PTEBS may have similar effects to the respective prototypes, which may improve the therapeutic efficacy of these molecules and decrease adverse effects in the long-term.
Assuntos
Glicemia/efeitos dos fármacos , Diabetes Mellitus Tipo 2/tratamento farmacológico , Glibureto/farmacologia , Hiperglicemia/tratamento farmacológico , Hipoglicemiantes/farmacologia , Pioglitazona/farmacologia , Animais , Relação Dose-Resposta a Droga , Glibureto/química , Homeostase/efeitos dos fármacos , Hipoglicemiantes/síntese química , Hipoglicemiantes/química , Resistência à Insulina , Estrutura Molecular , Pioglitazona/química , Ratos , Relação Estrutura-AtividadeRESUMO
We studied the effect and the mechanisms of action of 2α,3ß,23-trihydroxyolean-12-ene (THO), from Croton heterodoxus Baill. (Euphorbiaceae), in glucose uptake in hyperglycemic rats. The effect of in vivo pretreatment with THO in hyperglycemic rats was analyzed. The in vitro effects of THO were observed in adipocytes and in adipose tissue. THO reduced glycemia, in part by increasing serum insulin and augmenting the disposal of glucose as glycogen in hepatocytes but did not change the serum concentration of glucagon-like peptide-1. THO increased glucose uptake in adipocytes and in adipose tissue by a mechanism dependent on phosphatidylinositol 3-kinase vesicular traffic and on the process of vesicle fusion at the plasma membrane in regions containing cholesterol, indicating the involvement of glucose transporter-4 (GLUT4). This triterpene may act solely via the activation and translocation of GLUT4 (rather than via nuclear actions, such as upregulation of GLUT4 synthesis), since THO did not alter the amount of GLUT4 mRNA or the content of GLUT4. Consistent with these data, the stimulatory effect of this triterpene on the quantity of GLUT4 in the membrane fraction was dependent upon p38 phosphorylation. In this experimental model, orally administered 10 mg/kg THO did not modulate extracellular serum lactate dehydrogenase. In conclusion, THO decreases hyperglycemia by increasing serum insulin and hepatic glycogen content. The THO mechanism of action on adipose tissue for glucose uptake is suggested to be via GLUT4 translocation stimulation mediated by a p38-dependent mechanism. THO is a potential antihyperglycemic agent that acts in a target tissue for glucose homeostasis.
Assuntos
Insulina , Glicemia/metabolismo , Glucose , Homeostase/efeitos dos fármacos , Hipoglicemiantes/farmacologia , Insulina/metabolismoRESUMO
Erectile dysfunction is a common condition that affects men over age 40. It is highly related to obesity. The corpus cavernosum is the most important structure involved in erection. The aim of this study was to evaluate the structure of the corpus cavernosum of mice fed with a high energy density diet (HED). At 3 months of age, male C57BL/6 mice were fed with a HED diet (50% lipids) or standard chow (SC) diet (10% lipids) for 14 weeks. Afterwards, the animals were euthanized and the corpus cavernosum was analyzed through stereology. Statistical significance was calculated by the student's t-test (p < 0.05). The group fed with HED diet showed higher values of body weight, blood pressure and higher rates of cholesterol, triglycerides, and glucose from the second week to the end of the experiment. The HED group showed a significant increase in the connective tissue (15%) and a decrease in smooth muscle fibers (41%). The testosterone concentration in the HED group was 63% lower than in SC animals. Animals fed with a HED presented reduced testosterone serum levels and morphological changes on the corpus cavernosum, which may be related to erectile dysfunction.
Assuntos
Gorduras na Dieta/efeitos adversos , Miócitos de Músculo Liso/patologia , Pênis/patologia , Testosterona/sangue , Animais , Modelos Animais de Doenças , Disfunção Erétil/etiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Obesidade/fisiopatologia , Pênis/fisiopatologiaRESUMO
The goal of this study was to evaluate the influence of hypothyroidism induced by MMI, during gestation (G) or gestation plus lactation (GL) on testis and its relation with leptin in rats. Six to eight pups were killed at 90 days of age. For statistical analysis One-way ANOVA followed by the Holm-Sìdak post hoc test was used. Hypothyroidism resulted in a significant reduction in LH, FSH and testosterone and an increase in leptin serum levels (p < 0.04). There was a significant decrease in StAR, AR, FSHR, LHR, pSTAT3 and SOCS3 (p < 0.04) protein expression and in the fertility parameters (p < 0.04). We can conclude that hypothyroidism is associated with reduction of steroidogenesis and spermatogenesis leading to a low fertility potential in these animals. This outcome could be a consequence of low pituitary stimulus and testicular response and probably are not related with leptin hormone since its signaling pathway is down-regulated in the testis.
Assuntos
Hipotireoidismo Congênito/metabolismo , Hipotireoidismo Congênito/patologia , Hormônios/metabolismo , Testículo/patologia , Animais , Peso Corporal , Hipotireoidismo Congênito/sangue , Comportamento Alimentar , Feminino , Fertilidade/genética , Regulação da Expressão Gênica , Hormônios/sangue , Lactação , Masculino , Modelos Biológicos , Gravidez , Ratos Wistar , Espermatozoides/metabolismo , Testículo/metabolismoRESUMO
BACKGROUND: The effect of in vivo treatment with ursolic acid (UA) on glycemia in hyperglycemic rats and its mechanism of action on muscle were studied. METHODS: The UA effects on glycemia, glycogen, LDH, calcium and on insulin levels were evaluated after glucose tolerance curve. The ß-cells were evaluated through the transmission electron microscopy. UA mechanism of action was studied on muscles through the glucose uptake with/without specific insulin signaling inhibitors. The nuclear effect of UA and the GLUT4 expression on muscle were studied using thymidine, GLUT4 immunocontent, immunofluorescence and RT-PCR. RESULTS: UA presented a potent antihyperglycemic effect, increased insulin vesicle translocation, insulin secretion and augmented glycogen content. Also, UA stimulates the glucose uptake through the involvement of the classical insulin signaling related to the GLUT4 translocation to the plasma membrane as well as the GLUT4 synthesis. These were characterized by increasing the GLUT4 mRNA expression, the activation of DNA transcription, the expression of GLUT4 and its presence at plasma membrane. Also, the modulation of calcium, phospholipase C, protein kinase C and PKCaM II is mandatory for the full stimulatory effect of UA on glucose uptake. UA did not change the serum LDH and serum calcium balance. CONCLUSIONS: The antihyperglycemic role of UA is mediated through insulin secretion and insulinomimetic effect on glucose uptake, synthesis and translocation of GLUT4 by a mechanism of cross-talk between calcium and protein kinases. GENERAL SIGNIFICANCE: UA is a potential anti-diabetic agent with pharmacological properties for insulin resistance and diabetes therapy.
Assuntos
Glicemia/metabolismo , Cálcio/metabolismo , Insulina/metabolismo , Proteínas Quinases/metabolismo , Triterpenos/farmacologia , Animais , Cálcio/sangue , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Relação Dose-Resposta a Droga , Expressão Gênica/efeitos dos fármacos , Glucose/metabolismo , Transportador de Glucose Tipo 4/genética , Transportador de Glucose Tipo 4/metabolismo , Glicogênio/metabolismo , Hipoglicemiantes/farmacologia , Immunoblotting , Insulina/sangue , Insulina/farmacologia , Secreção de Insulina , Células Secretoras de Insulina/efeitos dos fármacos , Células Secretoras de Insulina/metabolismo , Células Secretoras de Insulina/ultraestrutura , L-Lactato Desidrogenase/sangue , L-Lactato Desidrogenase/metabolismo , Masculino , Microscopia Eletrônica de Transmissão , Estrutura Molecular , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/metabolismo , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Triterpenos/química , Ácido UrsólicoRESUMO
PURPOSE: We evaluated whether antihypertensive drugs that act through the renin-angiotensin system would affect testis function. MATERIALS AND METHODS: Ten mice were fed standard chow and 40 received a high energy density diet. At 8 weeks the high energy density diet mice were divided into 4 groups of 10 each. The untreated group received the high energy density diet alone. The 3 treated groups received that diet plus aliskiren (50 mg/kg daily), enalapril (30 mg/kg daily) and losartan (10 mg/kg daily), respectively, for the next 6 weeks. Blood pressure was measured twice monthly. At the end of the treatment period all mice were sacrificed. One-way ANOVA and the Holm-Sidak post hoc test were used to analyze results. RESULTS: The high energy density diet led to a significant increase in blood pressure (p <0.05). All treatments resulted in normalized blood pressure. In regard to reproductive function, and serum testosterone and estradiol the gene and protein expression of StAR, aromatase and luteinizing hormone receptor, and the protein expression of angiotensin-converting enzyme, renin and angiotensin type 1 receptor blocker were significantly decreased by the high energy density diet. Of the treatments only enalapril reverted the changes. Also, angiotensin-converting enzyme, angiotensin type 1 receptor blocker and renin protein expression were lower in all high energy density diet groups except the group that received enalapril. CONCLUSIONS: Only angiotensin-converting enzyme inhibitor reverted the hormonal and testis alterations caused by the high energy density diet. This suggests that enalapril should be the drug of choice for a patient who presents with previous reproductive dysfunction.
Assuntos
Inibidores da Enzima Conversora de Angiotensina/farmacologia , Enalapril/farmacologia , Fosfoproteínas/biossíntese , Sistema Renina-Angiotensina/efeitos dos fármacos , Testículo/efeitos dos fármacos , Testículo/metabolismo , Inibidores da Enzima Conversora de Angiotensina/uso terapêutico , Animais , Enalapril/uso terapêutico , Hipertensão/tratamento farmacológico , Hipertensão/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BLRESUMO
The leptin hormone is important to satiety and an important link between the nutritional status and reproductive processes. Owing to the contradictory effects of leptin on the ovary and the failure to clarify the precise mechanism by which leptin affects the ovary, our aim was to contribute to evaluation if leptin can directly regulate the gene expression of leptin itself and its receptors, and the expression of several genes related to the ovary function by a model of tissue culture. Ovaries from Wistar dams were used at 90 days of age and were submitted to medium with presence and absence of leptin. The results can demonstrate that leptin regulates gonadotropins and steroid receptors, which could suggest that the ovarian leptin role could be secondary to the changes in these receptors expression in rats.
La hormona leptina es importante en la sensación de la saciedad y un vínculo importante entre el estado nutricional y los procesos reproductivos. Debido a los efectos contradictorios de la leptina en el ovario y la falta de esclarecimiento del mecanismo exacto por el cual la leptina afecta el ovario, nuestro objetivo es contribuir a la evaluación si la leptina puede regular directamente la expresión del gen de la leptina sí mismo y sus receptores, y la expresión de varios genes relacionados con la función del ovario por un modelo de cultivo de tejidos. Los ovarios de las presas Wistar fueron usadas en los 90 días de edad y se sometieron a medio con presencia y ausencia de leptina. Los resultados pueden mostrar que la leptina regula las gonadotropinas y los receptores de esteroides, lo que podría sugerir que la función ovárica de la leptina podría ser secundario a los cambios en la expresión de sus receptores en ratas.
Assuntos
Gonadotropinas/metabolismo , Leptina/farmacologia , Ovário/efeitos dos fármacos , Ovário/metabolismo , Receptores de Esteroides/metabolismo , Animais , Feminino , Expressão Gênica/efeitos dos fármacos , Leptina/biossíntese , Leptina/genética , Reação em Cadeia da Polimerase , Ratos , Ratos Wistar , Receptores para Leptina/biossíntese , Receptores para Leptina/genéticaRESUMO
UNLABELLED: This paper aimed to evaluate the leptin role on the cellular proliferation and the expression of fibroblast growth factor 2, aromatase enzyme, and apoptotic genes in the human prostate tissue. METHODS: Fifteen samples of hyperplasic prostate tissue were divided in four symmetric parts maintained in RPMI medium supplemented with 10% fetal bovine serum, 1 ng/mL of gentamicin, and added with 50 ng/mL leptin (L) or not (C). After 3 hours of incubation, gene expression was evaluated by real time RT-PCR. Cellular proliferation was evaluated by immunohistochemistry for PCNA. RESULTS: The leptin treatment led to an increase cellular proliferation (C = 21.8 ± 0.5; L = 64.8 ± 0.9; P < 0.0001) and in the expression of Bax (C = 0.4 ± 0.1; L = 0.9 ± 0.2; P < 0.05) while Bcl-2 (C = 19.9 ± 5.6; L = 5.6 ± 1.8; P < 0.05), Bcl-x (C = 0.2 ± 0.06; L = 0.07 ± 0.02; P < 0.05), and aromatase expressions (C = 1.9 ± 0.6; L = 0.4 ± 0.1; P < 0.04) were significantly reduced. CONCLUSION: Leptin has an important role in maintaining the physiological growth of the prostate since it stimulates both cellular proliferation and apoptosis, with the decrement in the aromatase gene expression.
Assuntos
Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Leptina/farmacologia , Próstata/citologia , Próstata/efeitos dos fármacos , Humanos , Imuno-Histoquímica , Técnicas In Vitro , Masculino , Próstata/metabolismoRESUMO
The involvement of leptin in prostate diseases is related to an increase in the gene expression of both a and b isoform leptin receptors, leptin itself, androgen receptor, and aromatase, as well as by a reduction in both estrogen isoform receptors.