RESUMEN
AIM: This study investigated whether kisspeptin-10 (KP-10) prevents diabetic rhesus monkeys from insulin-induced hypoglycemic shock. MATERIALS AND METHODS: Thirty-six adult male rhesus monkeys were used, six in each group. Diabetes was induced with streptozotocin (45 mg/kg b.w.; single dose i.v.). Groups were as: saline control, insulin alone, pre-insulin (treated with KP-10, 30 min before insulin), post-insulin (treated with KP-10, 30 min after insulin), treated with premix dose of KP-10 (50 µg) and insulin, and the group treated with the kisspeptin antagonist P234 (50 µg). Following an overnight fast, each animal was subjected to respective treatment, and blood glucose concentrations were recorded every 30-min interval for 3 h. RESULTS: Intergroup comparisons demonstrated that treatment with KP-10 prior to insulin administration and kisspeptin-insulin premix treatment allowed blood glucose levels to rise to significantly higher levels (p < 0.001) by 180 min in diabetic and healthy animals compared to treatment with insulin alone. However, intragroup comparisons revealed a significant decrease in blood glucose level in diabetic animals only. Treatment with P234 antagonist followed by insulin administration abolished the preventive action of kisspeptin, whereby blood glucose decreased significantly (p < 0.001) in both diabetic and healthy animals. KP-10 post-insulin treatment, however, remained ineffective and led, instead, to significantly decreased glucose concentrations by 180 min in both diabetic and healthy animals when compared to animals treated with insulin alone. CONCLUSIONS: KP-10 bears therapeutic potential to prevent hypoglycemic shock that may sometimes occur during intensive insulin therapy. Several pharmacological aspects of its interaction with insulin and other drugs, however, remain to be investigated.
Asunto(s)
Hipoglucemia/inducido químicamente , Hipoglucemia/tratamiento farmacológico , Hipoglucemiantes/efectos adversos , Coma Insulínico/tratamiento farmacológico , Insulina/efectos adversos , Kisspeptinas/farmacología , Animales , Glucemia/efectos de los fármacos , Diabetes Mellitus Experimental/tratamiento farmacológico , Insulina/farmacología , Macaca mulatta , MasculinoRESUMEN
Effects of curcumin as antioxidant in extender were evaluated on freezability of buffalo spermatozoa. Semen from each of the five bulls (n = 3 replicates, six ejaculates/bull, a total of 30 ejaculates) was diluted in Tris-citric acid extender containing curcumin (0.5, 1.0, 1.5 or 2.0 mM) or control. At pre-freezing and post-thawing, total antioxidant contents (µM/L) and lipid peroxidation levels (µM/ml) were higher (p < .05) and lower (p < .05) respectively, with 1.5 and 2.0 mM compared to 0.5 and 1.0 mM curcumin and control. At post-thawing, progressive motility (PM, %) and rapid velocity (RV, %) were higher (p < .05) with 1.5 mM compared to other doses of curcumin and control (except in case of RV, 1.5 was similar with 1.0 mM). Kinematics (average path velocity, µm/s; straight-line velocity, µm/s; curved-line velocity, µm/s; straightness, %; linearity, %), in vitro longevity (%, PM and RV) and DNA integrity (%) at post-thawing were higher (p < .05) with 1.5 mM compared to control. At post-thawing, supravital plasma membrane integrity (%) and viable spermatozoa with intact acrosome (%) were higher with 1.5 compared to 2.0 mM curcumin and control. We concluded that freezability of water buffalo spermatozoa is improved with the addition of 1.5 mM curcumin in extender.
Asunto(s)
Crioprotectores/farmacología , Curcumina/farmacología , Preservación de Semen/métodos , Semen/efectos de los fármacos , Animales , Búfalos , Criopreservación , Daño del ADN/efectos de los fármacos , Masculino , Motilidad Espermática/efectos de los fármacos , Espermatozoides/efectos de los fármacosRESUMEN
The effects of equilibration times (E1, 2 h; E2, 4 h; E3, 6 h), freezing rates (FR1, manual, 5 cm above liquid nitrogen (LN2 ) for 10 min, plunging in LN2 ; FR2, programmable ultra-fast, holding at +4 °C for 2 min, from 4 to -10 °C at -10 °C/min, from -10 to -20 °C at -15 °C/min, from -20 to -120 °C at -60 °C/min, holding at -120 °C for 30 sec, plunging in LN2 ), and thawing rates (T1, 37 °C for 30 sec; T2, 50 °C for 15 sec; T3, 70 °C for 7 sec) were evaluated on quality of buffalo bull spermatozoa. Progressive motility (%), rapid velocity (%), average path velocity (VAP, µm/s), straight line velocity (VSL, µm/s), and mitochondrial transmembrane potential (%) were higher (p < 0.05) with E2, FR2, and T3 compared to other groups. Sperm curved line velocity (VCL, µm/s) was higher (p < 0.05) with E2 and FR2 compared to other groups. Sperm straightness (%) and linearity (LIN, %) were higher (p < 0.05) with E2 compared to other groups. Sperm LIN was affected (p < 0.05) with T3 compared to other groups. Supravital-plasma membrane integrity (%), viability and acrosome integrity (%) of spermatozoa were higher (p < 0.05) with E2 and FR2 compared to other groups. Sperm DNA integrity (%) was higher (p < 0.05) with FR2 and T1 compared to other groups. We concluded that inclusion of 4 h-equilibration time, programmable ultra-fast freezing rate, and rapid thawing at 70 °C for 7 sec in cryopreservation protocol improves the post-thaw quality of buffalo bull spermatozoa.
Asunto(s)
Criopreservación/métodos , Daño del ADN , Preservación de Semen/métodos , Motilidad Espermática/fisiología , Espermatozoides/citología , Animales , Búfalos , Congelación , MasculinoRESUMEN
The receptors for sulphonylurea (SURs) are known to be expressed in the mouse kidney, but their expression in the ureter is undefined. Owing to the physiological and pharmacological significance of SURs, the localization of SUR in ureters of adult mice and rats was investigated through immunohistochemistry. Animals were perfused transcardially with 4% paraformaldehyde and tissues were processed for immunohistochemistry using polyclonal antisera against SUR2A and SUR2B (SUR1) receptor proteins. Sections were incubated with primary and secondary antisera and developed with aminoethylcarbazole as a chromogen. A differentiated localized staining pattern of SUR proteins in rat and mouse ureters is demonstrated. In the mouse, immunoreactivity of SUR2A was predominantly confined to the cytoplasmic portion of epithelial cells and blood vessels, with comparatively low-level staining found in smooth muscle. In contrast, SUR2B (SUR1) immunoreactivity was absent in mouse ureters. In rats, SUR2A immunoreactivity was localized only in the blood vessels, while SUR2B (SUR1) immunoreactivity was localized in the epithelial cell cytoplasm. Tissue specificity of SUR is demonstrated in the two species of rodents and suggests a role of SUR proteins in urinary metabolism pertaining possibly to salt handling and maintenance of the smooth muscle tone.
Asunto(s)
Transportadoras de Casetes de Unión a ATP/metabolismo , Canales de Potasio de Rectificación Interna/metabolismo , Receptores de Droga/metabolismo , Uréter/metabolismo , Animales , Células Epiteliales , Regulación de la Expresión Génica/fisiología , Masculino , Ratones , Membrana Mucosa/metabolismo , Ratas , Ratas Wistar , Receptores de Sulfonilureas , Distribución TisularRESUMEN
The sulphonylurea receptor (SUR) is the site of action for sulphonylurea derivatives such as glibenclamide, which are widely used as oral hypoglycaemic agents. Sulphonylureas have also been shown to affect urine flow and salt excretion by the kidney; therefore, the use of these drugs may have important implications for the pharmacological manipulation of renal salt handling. The purpose of the present investigation was to increase our understanding of the possible role of SUR in the regulation of renal function by determining the distribution of SUR isoforms within mouse kidney. Immunostaining with anti-SUR antisera revealed specific staining of SUR2B in distal nephron segments of mouse kidney. A diffuse, low level staining was observed in proximal tubules in the inner cortical region. No evidence was found for the presence of SUR2B in intra-renal blood vessels. Reverse-transcription polymerase chain reaction and Western blotting experiments indicated that SUR2B is the only known isoform expressed. These data demonstrate that SUR2B in mouse kidney is expressed in tubule regions that are critical in determining renal salt excretion.