RESUMEN
OBJECTIVE: Observe the expression and distribution of HMGB1 in Sombati's cell model and kainic acid-induced epileptic rats' model. MATERIALS AND METHODS: Dissociated hippocampal neurons from neonatal SD rats and cultured those for 9 days, then changed medium to Mg2+-free medium for 3 hours to induce Sombati's cell model. The expression level of HMGB1 in the neurons was detected at 24h and 72h by Western Blotting. Appropriate kainic acid was injected into the lateral ventricles to induced epileptic rats' model in vivo trial, the expression level and distribution of HMGB1 at 24h and 72h were established by immunohistochemistry. RESULTS: The expression level of HMGB1 showed significantly different between model group and control group both in vitro and in vivo trials. At 24h, the expression level of HMGB1 in the model group was lower than the control group (p < 0.05), and became higher than the control group at 72h (p < 0.05). From the in vivotrial, a nucleus-to-cytoplasm translocation was also discovered. CONCLUSIONS: This investigation indicates that HMGB1 plays a crucial role in the pathophysiology of epilepsy, by altering its quantity and distribution.
Asunto(s)
Modelos Animales de Enfermedad , Epilepsia/metabolismo , Proteína HMGB1/biosíntesis , Ácido Kaínico/toxicidad , Animales , Animales Recién Nacidos , Células Cultivadas , Epilepsia/inducido químicamente , Epilepsia/patología , Regulación de la Expresión Génica , Proteína HMGB1/análisis , Hipocampo/química , Hipocampo/metabolismo , Hipocampo/patología , Masculino , Ratones , Neuronas/química , Neuronas/metabolismo , Neuronas/patología , Ratas , Ratas Sprague-DawleyRESUMEN
Solid- and solution-phase synthesis of peptidomimetic inhibitors of urokinase-type plasminogen activator based on the sequence dSerAlaArg-al are described. The biological activities of these unique inhibitors are reported herein. Carbonate prodrugs were prepared and tested as potential drug delivery systems.
Asunto(s)
Aldehídos/síntesis química , Dipéptidos/síntesis química , Inhibidores Enzimáticos/síntesis química , Activador de Plasminógeno de Tipo Uroquinasa/antagonistas & inhibidores , Aldehídos/farmacocinética , Aldehídos/farmacología , Animales , Área Bajo la Curva , Cromatografía Líquida de Alta Presión , Dipéptidos/farmacocinética , Dipéptidos/farmacología , Inhibidores Enzimáticos/farmacocinética , Inhibidores Enzimáticos/farmacología , Fibrinolisina/antagonistas & inhibidores , Semivida , Profármacos/síntesis química , Profármacos/farmacocinética , Profármacos/farmacología , Ratas , Activador de Tejido Plasminógeno/antagonistas & inhibidoresRESUMEN
An assay for the quantification of plasma and urine levels of CVS 1123, an orally bioavailable thrombin inhibitor, and its desmethyl form. CVS 738, was developed to support clinical and toxicology studies. This assay uses solid-phase extraction, reversed-phase HPLC separation, and post-column fluorescent derivatization with ninhydrin. An internal standard is added to correct for recovery. In aqueous solution, the arginine aldehyde structures of CVS 1123 and CVS 738 exist in multiple forms which can be separated under standard reversed-phase HPLC conditions. HPLC conditions were optimized to give rapid interconversion of the forms on the separation time scale, and consequently a single chromatographic peak. Extraction conditions were modified for quantitative extraction of drug compounds from large volumes of human plasma. The assay was shown to be accurate and precise, with a quantification limit of 17 ng CVS 1123/ml human plasma.
