RESUMEN
OBJECTIVE: The aim of our research was to analyze and compare the pharmacokinetics of paeoniflorin, calycosin, calycosin-7-o-ß-d-6-glucoside, and 6-gingerol in the blood and brain tissue of normal and cerebral ischemia-reperfusion injury rats by HPLC-MS/MS method. METHODS: The blood and brain tissue samples of normal and middle cerebral artery occlusion (MCAO) rats were compared. The blood and brain tissue samples were collected by using microdialysis technique. The concentrations of paeoniflorin, calycosin, calycosin-7-o-ß-d-6-glucoside, and 6-gingerol in blood and brain tissues were determined by the HPLC-MS/MS internal standard method. RESULTS: Compared with the normal group, the model group after the administration of the Huangqi Guizhi Wuwu Decoction showed that Cmax blood, AUC0-t blood, and AUC0-inf blood of paeoniflorin were increased, CLblood, t1/2 brain, and Vbrain of paeoniflorin were decreased; Cmaxblood, AUC0-tblood, AUC0-infblood, and average residence time (MRTbrain) of calycosin-7-o-ß-d-6-glucoside were decreased and the CLblood and Cmax brain of calycosin-7-o-ß-d-6-glucoside were increased; Cmax blood of calycosin was decreased, Vblood and Vbrain of calycosin were increased; Cmax blood, AUC0-t blood, AUC0-inf blood, and MRTbrain of 6-gingerol were decreased, CLblood of 6-gingerol was increased. CONCLUSION: This method is simple, rapid, and sensitive. It is suitable for the pharmacokinetic study of Huangqi Guizhi Wuwu Decoction in the blood and brain tissue of rats. Cerebral ischemia-reperfusion injury increased the content of paeoniflorin, calycosin, calycosin-7-o-ß-d-6-glucoside, and 6-gingerol in the blood, affecting the clearance rate of paeoniflorin in the brain, the detention time of calycosin-7-o-ß-d-6-glucoside and the 6-gingerol in the brain. In normal and cerebral ischemia-reperfusion rats, the content of paeoniflorin and 6-gingerol in the blood was higher than that in brain tissue, while the content of calycosin, calycosin-7-o-ß-d-6-glucoside in the brain tissue was higher than that in blood, suggesting that calycosin and calycosin-7-o-ß-d-6-glucoside have brain targeting properties.
Asunto(s)
Encéfalo/metabolismo , Medicamentos Herbarios Chinos/farmacocinética , Administración Oral , Animales , Cromatografía Líquida de Alta Presión , Medicamentos Herbarios Chinos/administración & dosificación , Medicamentos Herbarios Chinos/química , Masculino , Microdiálisis , Estructura Molecular , Ratas , Ratas Sprague-Dawley , Daño por Reperfusión , Espectrometría de Masas en TándemRESUMEN
Buyang Huanwu decoction (BHD), a popular formulation prescribed in traditional Chinese medicine (TCM) for the treatment of ischemic stroke, has been reported to have a potential role in promoting axonal regeneration. The purpose of the study was to screen and identify bioactive compounds from BHD using live PC12 cells coupled with high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Using this approach, we successfully identified six bioactive components from BHD. These components have protective effects on oxygen-glucose deprivation/reperfusion (OGD/R) injury to PC12 cells. Furthermore, calycosin-7-d-glucoside (CG) and formononetin-7-O-ß-d-glucoside (FG) could upregulate the protein expression of growth-associated protein 43 (GAP-43) and brain-derived neurotrophic factor (BDNF). This study suggests that living cells combined with HPLC-MS/MS can be used for the screening of active ingredients in TCMs.
Asunto(s)
Axones/efectos de los fármacos , Medicamentos Herbarios Chinos/farmacología , Regeneración/efectos de los fármacos , Animales , Axones/metabolismo , Isquemia Encefálica/tratamiento farmacológico , Isquemia Encefálica/metabolismo , Factor Neurotrófico Derivado del Encéfalo/metabolismo , Línea Celular Tumoral , Cromatografía Líquida de Alta Presión/métodos , Proteína GAP-43/metabolismo , Glucósidos/farmacología , Isoflavonas/farmacología , Medicina Tradicional China/métodos , Células PC12 , Sustancias Protectoras/farmacología , Ratas , Daño por Reperfusión/tratamiento farmacológico , Daño por Reperfusión/metabolismo , Accidente Cerebrovascular/tratamiento farmacológico , Accidente Cerebrovascular/metabolismo , Espectrometría de Masas en Tándem/métodosRESUMEN
Buyang Huanwu decoction (BHD) is a well-known traditional Chinese medicine that has long been used to treat ischemic brain damage which is associated with hemorheology. To screen active ingredients in BHD responsible for reducing blood viscosity by reducing red blood cell (RBC) lesions to treat ischemic stroke, a method involving RBC membrane binding and solid-phase extraction (SPE) was developed in this study. The components of BHD interacting with RBC were analyzed by mass spectrometry and four compounds, calycosin, paeoniflorin, 6-hydroxy behenol-3,6-di-O-glucoside and calycosin-7-O-ß-d-glucoside, showed binding affinity to RBCs. An erythrocyte activity assay revealed that the identified ingredients promoted the activities of Na+-K+-ATPase, sialic acid and superoxide dismutase and reduced the content of cholesterol on the RBC membrane, suggesting a mechanism underlying their anti-erythrocyte aggregation activity. Based on these results, the RBC membrane binding assay combined with SPE and mass spectrometry is a novel and effective approach for screening potentially anti-erythrocyte lesion constituents in traditional Chinese medicines.