RESUMEN
OBJECTIVE: To study the anti-portal hypertension effect of oleanolic acid (OA) in CCl4-induced cirrhosis rats and its mechanism. METHODS: Rats were induced to portal hypertension by CCl4. After treatment with low dose of OA (30 mg/kg) and high dose of OA (60 mg/kg) by intragastrically for a month, the parameters in serum or liver tissue including ALT, AST, MDA, GSH-Px, NOx, eNOS, cGMP and type I collagen were measured. The MAP, PP and HR were determined by hameodynamic method and the eNOS expression in liver was measured by western blot. The pathological changes of liver tissue were also tested by Masson dye. The normal group and model group were given 0.25% of CMC-Na solution. RESULTS: Compared with the model group, treatment with 30 mg/kg and 60 mg/kg OA significantly decreased the levels of ALT, AST, ALP, gamma-GT and MDA and enhanced the level of GSH-Px in liver (P<0.05). Moreover, the collagen content also notably lowered in CCl4-induced cirrhosis rats, thus decreasing the portal pressure (PP). However, the MAP and HR were not affected by OA treatment. In addition, the expression of eNOS in liver markedly increased after one mouth treatment of OA, hereof enhancing the level of cGMP and NOx in the CCl4-induced portal hypertensive rats (P<0.05). CONCLUSION: OA could inhibit the progress of fibrosis and lower the PP in CCl4-induced portal hypertensive rats and the anti-portal hypertension effect might be related to increasing the expression of eNOS and enhance the NOx level in liver.
Asunto(s)
Hipertensión Portal/tratamiento farmacológico , Cirrosis Hepática Experimental/tratamiento farmacológico , Óxido Nítrico Sintasa de Tipo III/metabolismo , Ácido Oleanólico/uso terapéutico , Fitoterapia , Sustancias Protectoras/uso terapéutico , Animales , Peso Corporal , Tetracloruro de Carbono/efectos adversos , Modelos Animales de Enfermedad , Hipertensión Portal/etiología , Hipertensión Portal/metabolismo , Hígado/efectos de los fármacos , Hígado/metabolismo , Hígado/patología , Cirrosis Hepática Experimental/inducido químicamente , Cirrosis Hepática Experimental/metabolismo , Pruebas de Función Hepática , Masculino , Malondialdehído/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo III/genética , Ácido Oleanólico/farmacología , Extractos Vegetales/farmacología , Extractos Vegetales/uso terapéutico , Sustancias Protectoras/farmacología , Ratas , Ratas Sprague-DawleyRESUMEN
A simple, rapid and sensitive liquid chromatography tandem mass spectrometry method is presented for the simultaneous determination of oleanolic acid, p-coumaric acid, ferulic acid, kaemperol and quercetin in rat plasma. Glycyrrhetinic acid was used as an internal standard, and sample pretreatment consisted of a liquid-liquid extraction. Chromatographic separation was achieved on a Gemini 110A C18 column (50 × 2.0 mm i.d., 5 µm) by gradient elution with a mobile phase consisting of methanol, acetonitrile and 0.01% formic acid in water. Tandem mass spectrometric detection was conducted using multiple reaction monitoring under negative ionization mode. Calibration curves offered linear ranges of two orders of magnitude with r > 0.99. The method was validated in terms of matrix effect, intra-day and inter-day precision, accuracy, linearity, specificity and stability. The relative standard deviation of intra-day and inter-day variations ranged from 2.66 to 14.74% and 1.9 to 14.55%. No substantial endogenous interference from blank plasma was observed. The method has been successfully applied to a pharmacokinetic study of Oldenlandia diffusa extract after oral administration in rats.
Asunto(s)
Cromatografía Liquida/métodos , Ácidos Cumáricos/sangre , Flavonoles/sangre , Oldenlandia/química , Ácido Oleanólico/sangre , Extractos Vegetales/farmacocinética , Animales , Ácidos Cumáricos/química , Ácidos Cumáricos/farmacocinética , Estabilidad de Medicamentos , Flavonoles/química , Flavonoles/farmacocinética , Modelos Lineales , Masculino , Ácido Oleanólico/química , Ácido Oleanólico/farmacocinética , Extractos Vegetales/administración & dosificación , Ratas , Ratas Sprague-Dawley , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Espectrometría de Masas en Tándem/métodosRESUMEN
A novel, simple, and sensitive method for the determination of jujuboside A in rat plasma using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) was developed. Following solid-phase extraction, measurement of jujuboside A was performed by negative ion electrospray ionization (ESI) in multiple reaction monitoring (MRM) mode. The limit of detection was 1.25 ng/mL, and the lower limit of quantification was 5 ng/mL in rat plasma. Good linearity was obtained over the range of 6.25-500 ng/mL, and the correlation coefficient was better than 0.998. The intra- and inter-day precisions ranged 4.4-7.5% and 2.9-10.7%, respectively. The accuracy derived from QC samples ranged 3.2-7.8% and 2.2-3.5%, respectively. The recovery ranged from 72.9 to 75.1% and the matrix effect from 96.7 to 105.3%. The analyte was stable under various conditions (at room temperature, during freeze-thaw, in the autosampler and under deep-freeze conditions). The developed method was successfully applied to the pharmacokinetic study in rats.