| Objective Study the representative of the effective parts composition of Astragalus and Ligusticum wallichii in cerebral ischemia injury in rat plasma pharmacokinetic and pharmacodynamics changes.Scientific evaluation of compatibility of traditional Chinese medicine(TCM)on cerebral ischemia model function and mechanism in animals.Providing a reference for Treating the disease of heart head blood-vessel and making Chinese medicine modernization.Methods(1)Study on Pharmacokinetics of the Effective part Compatibility of Astragalus and Ligusticum wallichii.①Building the methods of HPLC to detecting the concentration of Calycosin,TMP and Ferulic acid in rat plasma.The methanol volume change over time are as follows:0～5min,25%～25%;5～6min,25%～35%;6～10 min,35%～60%;10～14min,60%～60%.The column temperature is 30℃.Velocity of flow,1 mL·min-1.Determine wavelength,254 nm.②Using the orthogonal design method to divide the effective parts of Astragalus and Ligusticum wallichii to 9 groups.SD rats randomly divided into nine groups.And then set up middle cerebral artery occlusion(MCAO)model.Oral administration at the time reperfusion.Orbital blood 0.5 mL to 1.5 mL centrifuge tube which has been injecting heparin sodium before at 0.083、0.25、0.5、0.75、1.0、1.5、2.0、3.0、4.0、6.0 h after Oral administration.Using the methods of HPLC to detecting the concentration of Calycosin,TMP and Ferulic acid in rat plasma.Calculating pharmacokinetic parameters by DAS 3.2.6(2)Study on pharmacodynamics of the Effective part Compatibility of Astragalus and Ligusticum wallichii.SD rats randomly divided into nine groups.And then set up middle cerebral artery occlusion(MCAO)model.Oral administration at the time reperfusion.Orbital blood 0.5 mL to 1.5 mL centrifuge tube which has been injecting heparin sodium before at 0.083、0.25、0.5、0.75、1.0、1.5、2.0、3.0、4.0、6.0 h after Oral administration.Detection activation of LDH in rat plasma by Microplate reader.Results(1)Calycosin,TMP and Ferulic acid have good linearity range at 0.469～30μg·ML-1.2.5～100 μg·2mL-1.0.3125～20 μg·mL-1.And the Linear equation are as follows:Y=0.1273x-0.0479,R2=0.9984,Y=0.0023x+0.0009,R2=0.9991,Y=0.1066x+0.0054,R2=0.9994.The average recoveries rate of Calycosin are all above 90%,RSD<6%.The average recoveries rate of TMP are as follows:98.30%,RSD=3.02%;97.16%,RSD=7.65%;94.05%,RSD=8.74%.The average recoveries rate of Ferulic acid are as follows:98.08%,RSD=7.44%;92.81%,RSD=7.25%;93.0%,RSD=4.27%.The average precision of variation of day and day RSD were less than 10%,in line with biological sample analysis requirements.(2)Orbital blood 0.5 mL to 1.5 mL centrifuge tube which has been injecting heparin sodium before at 0.083、0.25、0.5、0.75、1.0、1.5、2.0、3.0、4.0、6.0 h after Oral administration.And then calculated the average drug concentration in plasma.TO get the main pharmacokinetic parameters by DAS 3.2.6.The pharmacokinetic parameters of Calycosin,TMP and Ferulic acid were assessed with noncompartment model.After oral administration.the pharmacokinetic curve of Calycosin are all appearing the double peak phenomenon.The Cmax of com 1、com2 and com4 are significant greater than the others.The T max of com1、com4 and com5 are significant less than the others.The AUC com2 and com6 are significant greater than the others.The MRT have no significant differences in rats after oral administration nine difference prescription medicines.They fundamental remained around 3.0 h.After oral administration nine difference prescription medicines.To get the pharmacokinetic curve of TMP by calculating,found that only com4 was one peak.The others are all two peaks.The Cmax of com2、com4、com6 and com7 are significant greater than the others.The T max of com3、com4 and com9 are significant less than the others.The AUC com6 are significant greater than the others,and next are com2、com4 and com7.The MRT fundamental remained 2.0～3.0 h after oral administration nine difference prescription medicines.Among them com 1、com6、com7 and com8 are approach to 3h,greater than the othersThe concentration of Ferulic acid in plasma are most reducing over time after oral administration nine difference prescription medicines.Coml、com7 and com9 appeared phenomenon of unimodal.The Cmax of com4 are significant greater than the others,and then was com4,the last were com3、com6 and com8.They values are 24.611 μg·mL-1,15.128 μg·mL-1,9.405 μg·-mL-1,9.364μg·mL-1 and 9.994 μg·mL-1.Com1、com7 and com9 reached Tmax at 0.5、0.25hand 0.167.The others reached Tmax around 0.083 h.Com7 have the biggest value of MRT in rats after oral administration nine difference prescription medicines.(3)Compared to normal group,the activation of LDH in rat plasma have significant increased after cerebral ischemia reperfusion injury(P<0.01).Compared to model groups the activation of LDH in rat plasma have varying degrees of decline at each time point after oral administration nine difference prescription medicines.It could give play to better pharmacological function before 2.0 h after cerebral ischemia reperfusion injury.Com3、com4、com5、com7 and com9 could significant reduce the activation of LDH in rat plasma(P<0.01).Conclusion(1)The condition of HPLC were convenient,reliable,sensitive and stable.The average recoveries were high and precision well.It’s apply to study Pharmacokinetics of Calycosin,TMP and Ferulic acid in rats plasma.(2)To analyze the pharmacokinetic parameters of Calycosin have found that com 1、com2 and com4 possess better absorption than the others after oral administration nine difference prescription medicines.The better bioavailability are com2 and com6 The MRT of Calycosin are all long.It indicated that it could remain longer pharmacodynamic effects in rats.The pharmacokinetic parameters of TMP showed that com3、com4 and com9 have faster rate of absorption than the athers.Com6 have the highest bioavailability,and then are com2、com4 and com7.Com1、com6、com7 and com8 could remain longer pharmacodynamic effects in ratsAfter analyze the pharmacokinetic parameters of Ferulic acid.We could find that com4 have the significant absorption than the others.It could be fast absorbed after oral administration nine difference prescription medicines.And it showed a downward trend over time.Com2、com3 and com4 have higher bioavailability than other groups.Com7 could remain the longest pharmacodynamic effects in rats(3)Compared to normal group,the activation of LDH in rat plasma have significant increased after cerebral ischemia reperfusion injury(P<0.01).Compared to model groups the activation of LDH in rat plasma have varying degrees of decline at each time point after oral administration nine difference prescription medicines.Coml could significant reduced the activation of LDH in rat plasma before 4 h after cerebral ischemia reperfusion injury.Coml could get treatment only at the time of 2 h fter cerebral ischemia reperfusion injury.Com6 and com8 could play better curative effect than after 1h after cerebral ischemia reperfusion injury.Com3、com4、com5、com7 and com9 could significant reduce the activation of LDH in rat plasma(P<0.01).They play more important role on treating cerebral ischemia reperfusion injury. |
PDF Full Text Request