Study On The Pharmacokinetics Of Eucommia Ulmoides-Salvia Miltiorrhiza Compatibility In Different Kidney Eficiency And Blood Stasis Models Rats And Identification Of Core Effector Substances

Kidney deficiency and blood stasis,as the root cause of aging and its complications,is also an important pathogenic factor inducing senile diseases.Eucommia ulmoides and Salvia miltiorrhiza were high-frequency listed in the prescription of Tonifying the kidney and activating blood circulation,and their compatibility mechanism and core effector substances need to be studied deeply.Guided by the idea of molecular traditional Chinese medicine,Aucubin（AU）,an active component for tonifying the kidney in Eucommia ulmoides and Danshensu（DSS）,an active component for promoting blood circulation and removing blood stasis in Salvia miltiorrhiza,were selected to study the pharmacokinetics and tissue distribution of the two compounds in rats with chronic and acute kidney deficiency and blood stasis model,The causal mathematical model is used to identify the core effector substances in plasma and clarify the effector substance group of the combined application of AU and DSS.The research contents are as follows:1.Study on the purification process of AUThe effects of different eluting alcohol solvent concentration and drying method were investigated,and the purity of AU was determined by HPLC-UV.The results showed that macroporous resin was eluted with water,2%,4%and 95%ethanol aqueous solution as eluent,The crude extract of Eucommia ulmoides seed meal was obtained by freeze vacuum drying,and the purity of prepared AU was 96.25%.2.Study on pharmacokinetics of AU and DSS in rats model of chronic kidney deficiency and blood stasis.An stable,reliable,specific and sensitive HPLC-UV method for the simultaneous determination of AU and DSS in rat plasma was established.The rats with chronic kidney deficiency and blood stasis were given AU and DSS aqueous solution respectively.Blood samples were collected at different time points before and after administration for analysis.Compared with the AU alone group,the half-life of elimination(T_1/2)of AU in the combined administration group was prolonged and the drug clearance rate（CLz/F）per unit time was significantly accelerated（P<0.05）,It shows that the combined administration not only prolongs the action time of the drug in the body,but also speeds up the clearance of the drug per unit time,so that the drug is not easy to accumulate in the body.Compared with DSS alone,the time to reach maximum concentration(T_max)（P<0.01）and mean residence time（MRT）（P<0.01）of DSS in the combined administration group were significantly prolonged.At the same time,DSS peaked twice after combined administration,indicating that AU and DSS cooperate with each other after combined administration,It can prolong the distribution time of AU and DSS in vivo,accelerate the drug clearance rate and promote the secondary absorption of DSS in vivo.3.Pharmacokinetics of AU and DSS in rats with acute kidney deficiency and blood stasis modelThe rat model of acute kidney deficiency and blood stasis was established by adrenaline combined with ice water bath.After giving AU and/or DSS,blood samples were collected at different time points for analysis.The results showed that compared with the single administration group,the t_1/2and AUC of AU in the combined administration group were increased,and the peak time of DSS increased(t_max).The AUC,MRT and t_1/2of Au and DSS in the chronic kidney deficiency and blood stasis model group were significantly higher than those in the acute kidney deficiency and blood stasis model group,while the CLZ/F was different,it shows that the clearance rate of AU and DSS were slowed down in the aging state,but this phenomenon can be significantly improved after combined medication.4.Study on the distribution of AU and DSS in organs of rats with different kidney deficiency and blood stasis models.After intragastric administration of,AU and/or DSS to rats models of different kidney deficiency and blood stasis,the rats were killed at 1 h,2 h and 3 h after administration,and the heart,liver,spleen,lung,kidney,brain,duodenum,uterus and ovary were collected.After treatment,the samples were injected and analyze by HPLC-UV.It was found that compared with the single administration,the combined administration promoted the distribution of AU and DSS in kidney,liver,heart The blood flow of spleen and other organs is rich,and it is better than that of single administration.In addition,after intragastric administration of AU and DSS,were distributed faster in the tissues of acute kidney deficiency and blood stasis model rats,and the concentration was higher than that of chronic kidney deficiency and blood stasis model rats,indicating that the utilization and metabolism of AU and DSS were slowed down in the elderly state,and significantly improved after combined medication.This phenomenon also preliminarily confirmed the results of pharmacokinetic study.5.Identification of core effector substances in rat plasma after combined application of Au and DSSBased on the synergy after the combined application of AU and DSS,the causal mathematical model was used to identify the core effector substances in the plasma of elderly rats after the combined application.A total of 34 pairs of mutually causal substance pairs were obtained,including 21 substances constituting the core effector group.