| Objective Qizhitongluo capsule is a kind of listed drug,which is used in the treatment of cerebral apoplexy and has a good curative effect.However Qizhitongluo capsule has many flavors and complex components,there is no study on the material basis of its efficacy.Therefore according to the efficacy of each drug in Qizhitongluo capsule,this study disassembled the prescription and studied the efficacy and material basis of Huoxuehuayu medicine after disassembling the prescription.Then the differences of the whole prescription,the Huoxuehuayu medicine,and the whole prescription deficiency Huoxuehuayu medicine were compared in the model rats of cerebral ischemia and reperfusion.Make clear the contribution of Huoxuehuayu Medicine Group to the whole prescription.To establish the fingerprint and serum fingerprint of Huoxuehuayu medicine group and to explore the material basis of the efficacy of Huoxuehuayu medicine group by comparing and analyzing the components entering blood.Methods 1.Rat model of cerebral ischemia-reperfusion was replicated by suture method.Qizhitongluo capsule group,Huoxuehuayu medicine group and ischemic Huoxuehuayu medicine group were administered intragastrically.The neurological function score,infarct size,NO,MDA,LDH,SOD,IL-1 ?,TNF-? in brain tissue were measured and judge protective effect on cerebral ischemia-reperfusion injury.2.To establish the UPLC fingerprint of drugs for Huoxuehuayu medicine.The common peaks of different batches of Huoxuehuayu drugs were found out and based on the UPLC atlas of single drug,deficient drug and reference substance under the same conditions,the common peaks were assigned and identified.So the contribution of each drug to the drug group was clarified.3.The fingerprint of blank rat serum and blank rat serum for intragastric administration was established by UPLC.Through the comparison of each chromatogram,the ingredientsthat entered the blood were found out.And to explore the possible material basis of Qizhuotongluo capsule for promoting blood circulation and removing blood stasis.Results 1.Compared with sham operation group,the neurological function score,infarct size and the contents of NO,MDA,LDH,IL-1 ? and TNF-? in brain tissue of rats in model group were significantly higher than those in sham operation group(P<0.05).The content of SOD decreased significantly(P<0.05).The results showed that the rat model of cerebral ischemia-reperfusion was successfully established.Compared with the model group,each administration group can reduce the neurological function score(P<0.05),improve the cerebral infarction(P<0.05),and reduce NO,MDA,LDH,IL-1?,TNF-? in brain tissue,increased the content of SOD in brain tissue(P<0.05).The results showed that each administration group had protective effect on cerebral ischemia-reperfusion injury in rats.Although the protective effect of Huoxuehuayu medicine group was not as good as that of the whole prescription(P<0.05),the protection level decreased after the whole prescription was short of Huoxuehuayu medicine(P<0.05).The protective effect of Huoxuehuayu medicine group was similar to that the whole prescription was short of Huoxuehuayu medicine group,which indicated that Huoxuehuayu medicine played an important role in protecting cerebral ischemia-reperfusion injury.It is of certain significance to study the effective substances of Huoxuehuayu drugs.2.UPLC,Zorbax SB-C18 column(100mm × 2.1mm,1.8?m),acetonitrile(A)-0.2%phosphoric acid water(B)(0~40min,5%~35%A;40~50min,35%~50%A),injection volume 1 ?L,detection wavelength 290 nm,column temperature 30 ?,flow rate 0.3m L·min-1,and signal acquisition time 50 minBy optimizing the preparation conditions and chromatographic conditions of the sample,the fingerprint of Huoxuehuayu medicine was established.The established method passed the methodological investigation.The fingerprint of different batches of Huoxuehuayu drugs was established by this method and the similarity between each batch was more than0.98,which proved that the method was stable,feasible and applicable.Twenty-one common peaks were identified.By comparing with the fingerprint of each single drug and drugs that lacks a specific flavor of Huoxuehuayu medicine,the common peaks were assigned.According to the fingerprint of the reference substance,it was identified that the4 th peak was protocatechuic acid,the 8th peak was caffeic acid,the 12 th peak was ferulic acid,the 14 th peak was rosmarinic acid,the 17 th peak was salvianolic acid B,and the 20 th peak was kaempferol.3.By analyzing the blood components of Huoxuehuayu drugs in the serum of blank rats,it was found that there were 11 blood entering components in blank rats,of which 6 wereabsorbed into blood directly in vitro and 5 were metabolized into blood.One of the six components absorbed directly into the blood was determined to be kaempferol.Conclusion The separation of Huoxuehuayu medicine from Qizhi Tongluo capsule to study the effective substance of Huoxuehuayu drug has certain significance to clarify the effective material basis of Qizhitongluo capsule.Through the establishment of fingerprint of Huoxuehuayu medicine in vitro,and the analysis of its blood entering components in blank rats may find some effective material basis for Qizhi Tongluo capsule.That can provide a theoretical basis for the quality control,scientific compatibility and clinical application of Qizhitongluo capsule. |
PDF Full Text Request