A Study On Preparation Of Taohong Siwu Decoction And Granules And Their Effects On Platelet Aggregation

Thrombosis is the basis of cardiovascular and cerebrovascular diseases.Antiplatelet aggregation drugs play a pivotal role in prevention and treatment of cardiovascular and cerebrovascular diseases.Currently,many kinds of antiplatelet drugs were used in clinical practice.Although these drugs can effectively inhibit the platelet aggregation induced by a certain platelet activator,there are many adverse reactions.(Such as allergic reactions,gastric mucosal lesion,coagulation dysfunction,and even the occurrence of recurrent ischemic events and the resulting increase in mortality).Taohong Siwu decoction,as one of the classical prescriptions for promoting blood circulation and dissipating blood stasis,can effectively improve platelet function.However,the decoction is not convenient to carry and the shelf life at room temperature is short,while granules are easy to carry and has a longer shelf life.Therefore,Taohong Siwu granules were prepared and the anti-platelet aggregation effect of granule and decoction was investigated in this thesis.The prescription of Taohong Siwu were pulverized,and Angelicae sinensis Radix and Chuanxiong Rhizoma were soaked in cold water for 0.5 hour and distillated,the distillate were preserved in a new container;The dregs were boiled with Persicae semen,Carthami Flos,Paeoniae Radix Alba,Rehmanniae Radix Praeparata for three times with distilled water.Each time the high heat is used until the water was boiling,and then gentle heat for another one hour.Filtrated,mixed extract and distilled liquid,and concentrated for Taohong Siwu Decoction.(Based on crude drugs is 0.5g/ml).The decoction continues to be concentrated to 1.0 g/m L based on the crude drug to obtain the extracts.The granules were prepared by mixing the extract with the auxiliary materials,and the forming process of granules was optimized by the particle forming rate and yield.The optimized particle molding process is as follows: the ratios of extract to starch,dextrin,and raw material powder are 9:3:2:2,respectively,and the air is fresh,blown by air blower at 60°C for 3 hours to dry and sieve.Thin-layer chromatography(TLC)for qualitative identification of Chinese herbal medicines,Ultra-high performance liquid chromatography was used to determine the content of amygdalin,ferulaic acid,dioxy-salicylic acid,peoniflorin,and Hydroxysafflower yellow A.The chromatographic fingerprint similarity evaluation software of traditional Chinese medicine was used to evaluate the ultra-high performance liquid chromatography of Taohong Siwu granules and decoction.The results showed that the thin layer identification method is highly specific and reproducible.Amygdalin,ferulaic acid,dioxy-salicylic acid,peoniflorin,and Hydroxysafflower yellow A were in the range of 7.5-120μg/ml,1.25-20μg/ml,1.25-20μg/ml,10-160μg/ml,and 5.0-80μg/ml,respectively.The linear relationship between peak area and concentration was good.The recoveries of the spiked samples were 100.1%,102.1%,100.2%,99.88% and 98.18%,respectively.The content determination method is highly efficient,accurate,and highly operable.The similarity between the granules and the decoction chromatogram is above 0.9.On this basis,the anti-platelet aggregation effect was also researched.Adenosine diphosphate(ADP)and Arachidonic acid(AA)were used to induce platelet activation.Platelet aggregation rate was detected by nephelometry.SD rats were randomly divided into 5 groups---Blank control group,Taohong Siwu decoction group,Taohong Siwu granules group,ADP positive drug group(Ticlopidine hydrochloride group),AA positive drug group(acetosalicylic acid group).Each group of animals was continuously fed for 3 days once a day.After 2 hours of the last administration,blood was collected from aorta abdominalis(natrium citricum 1:9 for anticoagulant).Platelet-rich plasma and platelet-poor plasma were separated,and platelet-poor plasma-diluted platelet-rich plasma was diluted to 300×10~9 cells/L.ADP and AA were used as inducers to detect the maximum aggregation rate of platelets in each group,to calculate the inhibition rate of drug on platelet aggregation in each group and to evaluate the effect of Taohong Siwu granules and decoction on platelet aggregation.ADP induction results showed that compared with the blank control group,ADP positive drug group significantly reduced ADP induced platelet aggregation(p<0.05),Taohong Siwu granules group and Taohong Siwu decoction group significantly reduced ADP-induced platelet aggregation(p<0.05),AA-positive drug group had no statistical significance for ADP-induced platelet aggregation(p>0.05);AA induction results showed that AA-positive drug group significantly reduced AA-induced platelet aggregation compared to the blank control group(p<0.05),Taohong Siwu granules group and decoction group had a tendency to reduce AA-induced platelet aggregation,and ADP-positive drug group had no statistical significance for AA-induced platelet aggregation(p>0.05).