Capture of anti-coagulant active ingredients from Moutan Cortex by platelet immobilized chromatography and evaluation of anticoagulant activity in rats.
Moutan Cortex (MC) is a well-known Chinese medicine for promoting blood circulation and relieving blood stasis. The intent of this study was to evaluate the anticoagulant activity of MC and capture the bioactive compounds by platelet immobilized chromatography. Sprague Dawley (SD) rats were randomly divided into the control group, aspirin group and MC group (1.25, 2.5, 5g/kg/d). Coagulation system and platelet activity were investigated to evaluate the anti-coagulation effect of MC. The effective components of MC were captured by platelet immobilized chromatography. High performance liquid chromatography-diode array detection (HPLC-DAD) and liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) analysis were used to identify the binding ingredients. Meanwhile, the efficacy of active ingredients was assessed through inhibiting platelet adhesion and regulating the expression of platelet related proteins. Principal findings showed that 2.5g/kg/d MC significantly prolonged thrombin time (TT) and 5g/kg/d MC significantly prolonged TT and prothrombin time (PT). MC exhibited an inhibitory potency on adenosine diphosphate-induced platelet aggregation. Four active compounds were found by platelet immobilized chromatography including oxypaeoniflorin, tetragalloylglucose, pentagalloyl glucose and benzoylpaeoniflorin; these active ingredients significantly up-regulated the expression of hsp-70 and coronin-1B, reduced the ratio of adhesion platelets. These results suggest that MC markedly promoted blood circulation and relieved blood stasis by inhibiting platelet activation, as an anti-coagulant, elucidating its potential capacity to treat cardiovascular diseases.
['Qiu|Huihui|H|', 'Zhang|Li|L|', 'Zhu|Maomao|M|', 'Zhang|Minghua|M|', 'Chen|Juan|J|', 'Feng|Liang|L|', 'Jia|Xiaobin|X|', 'Jacob|Joe Antony|JA|']
D000818:Animals D000925:Anticoagulants D001792:Blood Platelets D002851:Chromatography, High Pressure Liquid D004305:Dose-Response Relationship, Drug D004353:Drug Evaluation, Preclinical D004365:Drugs, Chinese Herbal D008297:Male D029596:Paeonia D010936:Plant Extracts D011897:Random Allocation D051381:Rats D017207:Rats, Sprague-Dawley D021241:Spectrometry, Mass, Electrospray Ionization D053719:Tandem Mass Spectrometry
Anti-coagulant Moutan Cortex Platelet immobilized chromatography HPLC–MS/MS