Study On Correlation Of Integrated Pharmacokinetics And Pharmacodynamics Of Moutan Cortex In Blood-Heat Rats

Objective:A method of UPLC-MS/MS was established for simultaneous determination of 14 index components in the MC,to study the integrated pharmacokinetics and pharmacodynamics of the multi-index components of MC in the rat model of blood-heat（BH）,the"quantity-time-effect"curve was used to described and predicted the concentration-effect process of multi-index components in the MC,to analyze the correlation between integrated pharmacokinetics and pharmacodynamics（PK-PD）of multi-index components in the MC,and it provides methods and ideas for the basic research of pharmacodynamic substances of MC.Methods:（1）The powder of MC was extracted with 90%ethanol reflux,and the ultra performance liquid chromatography/tandem mass spectrometry（UPLC-MS/MS）method was used to determine the content of 14 index components in the MC.（2）The UPLC-MS/MS method was used to detect the 9 index components of alcohol extract from MC in plasma of rats,and the study on integrated pharmacokinetics of MC in rats under BH was established to investigate the pharmacokinetic behavior of MC under BH state.（3）Established a rat with BH model,then the body temperature of rats in the blank group,BH model group and administrated with MC from Bozhou and Tongling group were measured,calculated the temperature response index at 8 hours（TRI₈）after administration of the rats,in addition,the four indexes of rats’s blood rheology,coagulation and the inhibitory strength of MC on inflammatory factor TNF_αwere also measured,established a time-effect curve to investigate the pharmacodynamic characteristics of MC from Tongling and Bozhou.（4）The correlation analysis was conducted between the pharmacodynamic intensity-time change of TNF_αinhibition and the integrated plasma concentration-time change of multi-index components.Results:（1）Using UPLC-MS/MS to detect 14 chemical constituents（benzoic acid,benzoylpaeoniflorin,benzoyloxypaeoniflorin,paeonol,apiopaeonoside,paeonolide,catechin,mudanpioside C,gallic acid,methyl gallate,paeoniflorin,albiflorin,pentagalloylglucose and oxypaeoniflorinin）in the MC,simultaneously,a total of 10batches of medicinal materials from Tongling and Bozhou were tested for chemical composition.Through comparison of various ingredients,it was found that the content of phenols and phenol glycosides（paeonol,apiopaeonoside and paeonolide）in the MC from Tongling was significantly higher than that in Bozhou（P<0.01）,there was no significant difference in other components of MC between the two places.（2）Through pharmacokinetic experiments,a method was established to simultaneously detect 9 main chemical components（benzoic acid,benzoyloxypaeoniflorin,paeonol,apiopaeonoside,paeonolide,paeoniflorin,albiflorin,pentagalloylglucose and oxypaeoniflorin）of MC in rat plasma by UPLC-MS/MS,the integrated pharmacokinetic parameters AUC_（0-∞）of MC from Tongling and Bozhou in BH model rats are 280.13±42.57μg·mL^-1·h,60.16±14.34μg·mL^-1·h,MRT_（0-∞）are6.87±0.33 h,4.56±0.62 h,t_1/2are 6.87±3.33 h,1.34±0.22 h,T_maxare 4.00±0.00 h,6.00±0.00 h,C_maxare 38.92±3.20μg·mL^-1,11.15±3.33μg·mL^-1;the AUC_（0-∞）of monoterpene glycoside components（benzoyloxypaeoniflorin,paeoniflorin,albiflorinand oxypaeoniflorin）)are 23.80±1.21μg·mL^-1·h,23.80±1.67μg·mL^-1·h,MRT_（0-∞）are 7.68±0.12 h,5.22±0.34 h,t_1/2are 3.49±0.47 h,1.64±0.11 h,T_maxare4.00±0.00 h,4.00±0.00 h,respectively,C_maxare 4.49±0.023μg·mL^-1and 3.68±0.065μg·mL^-1,respectively.（3）The results of pharmacodynamic experiments show that the body temperature response index（TRI₈）at the 8th hour obtained by model group,Tongling MC administration group and Bozhou MC administration group werere 2.6,0.7,1.3℃·h^-1,spectively;by the coagulation four indices and hemorheological examinations showed that the model group could significantly prolong APTT and hemorheological examinations（P<0.05）.Both MC from Tongling and Bozhou can reduce the hemorheological examinations and APTT to a certain extent（P<0.05）,there are no significant differences between the four coagulation and hemorheological examinations of MC.A PK-PD model that integrates pharmacokinetic data and the inhibitory strength of MC against the inflammatory factor TNF_αwas established.MC from Tongling and Bozhou exhibited delays in drug efficacy,with delays of 120 min and 90 min,respectively.Conclusion:（1）Compared with MC from Bozhou,the content of main index components such as paeonol,paeoniflorin,pentagalgol glucose,paeoniflorin glycoside etc.in MC from Tongling was higher,and the integration of pharmacokinetic parameters AUC_（0-∞）,C_max,MRT_（0-∞）also showed the advantages of genuine medicinal materials.（2）The blood coagulation,hemorheology and APTT of BH model rats were improved to a certain extent in both areas,but there was no significant difference between the two,showed that MC achieves the effect of cooling blood by improving the hemorheology and preventing abnormal function of endogenous coagulation factors,the results of temperature measurement,the results of temperature measurement,pharmacodynamic index TRI₈and inhibitory effect of TNF_αall indicated that the MC in Tongling had a better effect of clearing heat and cooling blood than that in Bozhou.（3）The established PK-PD model of MC suggests that MC in Tongling and Bozhou inhibit the TNF_αintensity-time curve and the integrated plasma concentration-time curve tend to be consistent,both of which have a delay in drug efficacy,and the effect of MC in Tongling is delayed for a longer time.