Studies Of Serum Pharmacology,Serum Fingerprint And The Main Components Plasma Pharmacokinetics Of Traditional Mongolian Medicine Flos Scabiosae

Objective To determine the maximum dose of alcohol extract of Flos Scabiosa;To study the hepatoprotective effect of Flos Scabiosaes;To explore the hepatoprotective components basis of Flos Scabiosaes;To speculate the active chemical components of protecting liver,and screen quality marker（Q-marker）candidates of Flos Scabiosaes.Methods And Results（1）Acute toxicity test study:The classical acute toxicity test method was used to evaluate the toxicity of Flos Scabiosae alcohol extract and it was found that the mice did not died after the maximum concentration and volume administration;Thus,the half lethal dose was unable to speculate.The maximum administration dose was 198.88g（raw drug）/kg,which was equal to 145.70 times of the clinical equivalent dose of adult.It was considered that the alcohol extract of Flos Scabiosae was safe at this dose.（2）Serum pharmacology study of Flos Scabiosaes:The Flos Scabiosaes serum pharmacology evaluation method was adopted,induced the hepatocyte oxidative damage model by H₂O₂,and the cell viability was detected by MTT method at different time points.The results showed that except for 4hours and 6hours of drug-containing serum,the survival rate of hepatocytes in the drug-containing serum group was significantly increased at the other time points（P<0.001）,and the survival rate of liver injury model was the highest within 1.5hours drug-containing serum.This study indicated that Flos Scabiosaes drug-containing serum had a protective effect on H₂O₂-induced liver injury cells.（3）Establishment of serum fingerprint of Mongolian medicine Flos Scabiosae:The method of serum fingerprint analysis method was established by HPLC-Q-Exactive MS.The Fifteen batches of drug-containing serum of Flos Scabiosae were prepared by intragastric administration with abnormal Wistar rats as donors,and the above-mentioned analytical method was used to establish the fingerprint of drug-containing serum.After the data was preprocessed,the pattern recognition and multidimensional statistical analysis was calculated by SIMCA-P12.0software,and unsupervised principal component analysis（PCA）,the results showed that fifteen batches of medicinal materials aggregated well compared with the blank group,which indicated that the similarity between the batches of medicinal materials was good under the common mode;The systemic cluster analysis（HCA）was used to classify the serum fingerprints of 15 batches of medicinal materials into three categories;Through the relevant literature reports and standard reference comparison,a total of 18 prototypes were identified.（4）Exploring the hepatoprotective basis of FS based on time-spectrum-effect relationship:Based on serum pharmacological results and serum fingerprinting analysis method,the correlation between the survival rate of oxidatively injured hepatocytes and the peak area of each component of drug-containing serum at different time points was analyzed by grey correlation method.The results showed that the correlation between the 13 components and the liver protection activity was greater than 0.75,indicating that these components had different degrees of liver protection,and the components with correlation degree greater than 0.8 included 3,5-dicaffeoylquinic acid p-counmaric acid,diosmetin,apigenin and p-hydroxybenzoic acid,it was speculated that these components may exhibit relatively high hepatoprotective activity.（5）A comparative pharmacokinetics study of 13 components of FS in normal and acute liver injury states:Serum fingerprint analysis was applied to the pharmacokinetic comparison of 13 components after administration in normal and CCL4-induced acute liver injury in rats.The results showed that acute liver injury could significantly affect the absorption and metabolism of various components of Flos Scabiosaes,and the trends of pharmacokinetic parameters of different components had certain differences.Compared with normal rats,the maximum plasma concentration(C_max)and the area under the time curve（AUC）in model rats of 3,5-dicaffeoylquinic acid,neochlorogenic acid,p-coumaric acid,p-hydroxybenzoic acid,protocatechuic acid and aesculin were higher than those of normal rats,which indicated that the amount exposure of model rats was increased,and the plasma drug concentration of these components could be increased under the condition of liver injury,and the desired drug effect can be obtained at a lower dose.These components were supposed to be hepatoprotective active ingredients.Conclusion The maximum dosage of Flos Scabiosaes alcohol extract was determined,and the hepatoprotective effect of drug-containing serum of Flos Scabiosaes alcohol extract was confirmed,and fifteen batches of Flos Scabiosaes serum fingerprints were established to reflect the absorption of Flos Scabiosaes into blood after oral administration,and the Spectrum-Effect relationship of hepatoprotective activity was analyzed by grey correlation method.The difference of pharmacokinetic behavior of Flos Scabiosaes prototype components in normal-acute liver injury was clarified.This study laied a foundation for elucidating the research of pharmacodynamic substances in Flos Scabiosaes,and provided reference for the determination of quality marker（Q-marker）candidates of Flos Scabiosaes and clinical drug safety,which provided a scientific basis for Flos Scabiosaes modernization research,and promoted the deep development and utilization of this medicinal material.