Effects of Buyang Huanwu decoction and Astragalus mongholicus on platelet activating factor receptor activity in rabbits in vitro

摘要

BACKGROUND:The pharmacological action of traditional Chinese medicine compound is the comprehensive effect of the various ingredients,and the interactions of various ingredients are closely correlated with the final effect.In order to reveal the compatibility mechanism of BHD's prescription in treating and preventing ischemic cerebrovascular disease,we needed explore the effect and relation of ingredients in the prescription.OBJECTIVE:To observe the effect of Buyang Huanwu decoction(BHD)and Astragalus mongholicus on the activity of platelet activating factor receptor(PAFR)in the platelet of rabbits in vitro,and investigate the mechanism of Astragalus mongholicus.DESIGN:A decomposed recipes study.SETTING:Guangzhou University of Traditional Chinese Medicine.MATERIALS:Five New Zealand rabbits,weighing 2-3 kg,both sexes,were used.BHD was composed of Sheng Huang Qi 120 g,Dang Gui Wei 6 g,Chi Shao 4.5 g,Chuan Xiong 3 g,Di Long 3 g,Tao Ren 3 g,Hong Hua 3 g.The prescription for activating blood circulation consisted of Dang Gui Wei 6 g,Chi Shao 4.5 g,Chuan Xiong 3 g,Di Long 3 g,Tao Ren 3 g and Hong Hua 3 g.The prescription for invigorating qi consisted of 120 g Sheng Huang Qi.The prepared herbal pieces were purchased from the traditional Chinese medicine Dispensary of Foshan Second People's Hospital,and appraised by Professor Xu from Science of Chinese Materia Medica College,Guangzhou University of Traditional Chinese Medicine.3H-PAF was supplied by Amersham Co.,Ltd.(specific activity:6.475 TBq/mmol;batch number:200402);PAF standard by Biomol Co.,Ltd.(batch number:P1318V).METHODS:The experiments were carried out in the Laboratory of Nuclear Medicine,Guangzhou University of Traditional Chinese Medicine from September to December 2004.①Injections of BHD,prescriptions for activating blood circulation and invigorating qi were prepared by the decoction and alcohol sedimentation technique.Rabbit common carotid artery blood(40 mL)was drawn via intubation to prepare platelet suspension of (0.8-1.0) ×1010L-1.②Determination of 3H-PAF and washed PAFR binding:The general combination tube(T) contained washed platelet-rich plasma(WPRP)380μL +3H-PAF(0.35 nmol/L)10μL+distilled water 5μL:The nonspecific binding tube(P)contained WPRP 380μL+3H-PAF(0.35 nmol/L)10μL+cold PAF(1μmol/L)5μL:The sample tube(Y)contained WPRP 380μL+3H-PAF(0.35 nmol/L)10μL+experimental medicine(injection of BHD,prescriptions for activating blood circulation or invigorating qi)5μL.The test was conducted for three times for each sample in the same way as mentioned above.The samples were shaken on the oscillator for 30 s,then bathed at 25℃ for 40 minutes,and the reaction was terminated with cold Tris buffer containing 0.1% BSA.multichannel cell detachment separator was used for vacuum suction to filter the separated free 3H-PAF,and the filter paper was washed with cold Tris buffer for four times,then dried in the baking oven(80℃)for 1 hour,and placed in xylol liquid scintillator,and the radioactivity was determined automatically by the liquid scintillation detector.The mean of the three parallel tubes was calculated.The specific binging inhibition rate was calculated:SBIR=[(T-Y)/(T-P)]×100%].③Univariate analysis of variance was conducted.And for comparison of each paired groups,the q test was adopted.MAIN OUTCOME MEASURES:Effect of BHD whole prescription,prescriptions for activating blood circulation and invigorating qi on the specific binding inhibition rate of 3H-PAF and PAFR.RESULTS: BHD,prescriptions for activating blood circulation and invigorating qi were all able to inhibit the specific binding of 3H-PAF to PAFR,the specific blinding inhibition rates were(45.90±7.50)%.(97.90±1.84)% and(26.75±2.48)%,respectively,and there were significant differences between every two groups(P＜0.01).CONCLUSION:Single Astragalus mongholicus(120 g)can inhibit the specific blinding of PAFR in the platelet of the rabbit with 3H-PAF,but the combination of Astragalus mongholicus with the drugs for activating blood circulation in BHD can significantly decrease the inhibiting action of the latter on PAFR activity of the platelet,reflecting the combined mechanism of "removing blood stasis without injuring the vital qi"in BHD.