Study On Dengzhanxixin Injection And Its Caffeoylquinic Acids For Aspirin Resistance Prevention

Currently,aspirin(ASA)has been commonly used as a part of the initial clinic therapy for cardio-cerebrovascular diseases owing to its excellent potency ratio.However,the high incidence of ASA resistance happened on patients,which did not produce the expected antiplatelet effect.Salicylic acid(SA),the main metabolite of ASA,is now considered as the major reason of ASA resistance occurrence.Dengzhanxixin injection(DI)has the practical efficacy of promoting blood circulation to remove stasis blood,relieving rigidity of muscle and activating collaterals,which often combined with ASA for the treatment of cardio-cerebrovascular diseases and has obtained the good curative effect.The subject proposed whether DI could improve ASA resistance by ASA esterase system in vitro,platelet aggregation test in vitro,and pharmacokinetic and pharmacodynamic experiment in vivo.Our project would provide a new methodology for developing the therapy of herbal medicine for ASA resistance prevention.To investigate the effects of chlorogenic acid and its isomers on the activity of ASA esterase in human plasma.The concentrations of ASA and its metabolite SA in the culture in vitro were determined by HPLC.The ratio of CSA/(CASA+CSA)was used as the activity of ASA esterase.The effects of chlorogenic acid and its isomers on ASA esterase were evaluated by comparing the activities of ASA esterase in human plasma between blank control and experimental group.The results suggested that there were no significant differences between the blank control and experimental group.Chlorogenic acid and its isomers had no significant effect on the activity of ASA esterase at the experimental concentrations.To investigate the effects of dicaffeoylquinic acids on the activity of ASA esterase in human plasma.The activity of ASA esterase was determined by HPLC.The effects of dicaffeoylquinic acids on ASA esterase were evaluated by comparing the activities of ASA esterase in human plasma between blank control and experimental group.And homology modeling and molecular docking were employed to simulate their interaction.There were no significant differences between the blank control and experimental group,and the results of molecular docking indicated better inhibitory effect of 1,3-dicaffeoylquinic acid on ASA esterase binding.1,3-dicaffeoylquinic acid showed only a slight inhibition of ASA esterase,as well as 3,4-dicaffeoylquinic acid and 3,5-dicaffeoylquinic acid had no significant effect on the activity of ASA esterase.To focus on the effects of caffeoylquinic acids on ASA resistance in vitro.The inhibitive rate of caffeoylquinic acids on platelet aggregation in rabbits induced by arachidonic acid(AA)in vitro.Combined use of cafFeoylquinic acids、ASA and SA showed inhibitory effects on the platelet aggregation induced by AA in vitro.Caffeoylquinic acids could reverse aspirin resistance induced by SA in vitro.To develop a rapid and sensitive HPLC method to determine SA in rat urine,and study the effect of DI on excretion of SA in rat.The SD rats were divided into single and combined administration groups.The combined group was injected with DI before orally treated with ASA,while the single group was received 10 mg/kg oral administration of ASA.The concentration of SA in urine was determinated by HPLC method to compare the difference of excretion of SA under two administration models.The determination method with good precision and stability was suitable for the assay of SA in the urine.The differences of excretion of S A under single and combined administration models were significant.DI could accelerate the excretion of SA in rats.To study on pharmacokinetic and pharmacodynamic interaction of ASA and DI in rats.Six groups of six rats each were used according to a parallel study.Group A and group D(ASA)received daily repeated 10 mg/kg oral administration of ASA.Group B(DI)received daily repeated 4 mL/kg intravenous administration of DI.Group C and group E(ASA+DI)were administered as drug combination,namely daily dose of 10 mg/kg ASA and 4 mL/kg DI.Group F was blank control group.Group A,B and C were for pharmacokinetic study,then group D,E and F were for pharmacodynamic study.Both SA and scutellarin showed a two-compartment model pharmacokinetic profile.The concentration-time course of scutellarin was not changed by ASA,but the pharmacokinetic parameters of SA were changed by DI.Co-administration of ASA and DI decreased the TXB2 in rats.This study indicated that co-administration of ASA and DI can result in an apparent herb-drug pharmacokinetic and pharmacodynamics interaction in rats.DI can prevent ASA resistance.