Metabolomics Study Of Danshen Combined With Chuanxiong Against Focal Cerebral Ischemia

ObjectiveDanshen and Chuanxiong is a commonly used effective drug pair for the treatment of focal cerebral ischemia.Metabolomics has a holistic feature that helps evaluate the intervention of Danshen and Chuangxiong on focal cerebral ischemia.Therefore,this subject intends to conduct a metabolomics study of Danshen and Chuanxiong intervention on focal cerebral ischemia model.The focal cerebral ischemia model was replicated by filament method,neurobehavioral scoring and brain tissue staining were used to assess the reliability of the model.This experiment was conducted on the basis of screening qualified model rats for metabolomics study of Danshen and Chuanxiong on interventional focal cerebral ischemia model.The rat plasma and brain tissue samples metabolomics study method was established based on.Based on this model,a metabonomic study of Danshen and Chuanxiong on the intervention of focal cerebral ischemia was carried out.And then,based on this model,a metabolomics study of Danshen and Chuanchuan medicine on cerebral ischemia was performed.Based on metabolomics,the overall evaluation of the effects of Danshen and Chuanchiong medicine on interventional model of focal cerebral ischemia.Finally,the dynamic characteristics of the cerebral ischemic effect site of its component group are obtained,which will help to provide reference for subsequent research and clinical rational application.Methods1 Extraction,lyophilization and quality control of lyophilized products of Danshen and Chuanxiong decoctionThe lyophilized product was obtained by decoction and vacuum freeze-drying method,and the HPLC method for the determination of Danshensu,salvianolic acid B and ferulic acid in the lyophilized extract of Danshen and Chuanxiong was established.2 Replication and evaluation of focal cerebral ischemia model The model of focal cerebral ischemia was replicated by filament method,and the neurobehavioral scores of the model rats were scored by Longa’ s five-level four-point method after 2h and 24h ischemia,at the same time,the reliability of the model was further evaluated by rat brain TTC staining.3 Metabolomics study of rat plasma and brain tissue based on UPLC-Q/TOF-MS UPLC-Q/TOF-MS analysis methods for rat plasma and brain tissue samples were established.PCA and OPLS-DA metabolomics models were established to elucidatethe differences in plasma and brain tissue metabolomics.4 Metabolomics study of rat striatum based on UPLC-Q/TOF-MS The focal cerebral ischemia model was replicated by filament method,and brain microdialysis sampling technique was used to dentify differential metabolite.Results1 Extraction,lyophilization and quality control of lyophilized products of Danshen and Chuanxiong decoctionAccording to the dosage of Chinese Pharmacopoeia,the ratio of Danshen and Chuanxiong(15:10)was used.After extraction and vacuum freeze-drying,yellow-brown loose porous lyophilizate was obtained,the content of Danshensu,salvianolic acid B and ferulic acid was determined by HPLC.The Danshensu,Salvianolic acid B and ferulic acid average content was respective 1.612mg/g,31.301mg/g and 1.216 mg/g.2 Replication and evaluation of focal cerebral ischemia model The focal cerebral ischemia model was replicated by the filament method,and the model success rate was 98%,and the model 7-day survival rate was 50%.Neurobehavioral scores were performed at 2h and 24h after model establishment and replication 80%of which was 1 point,which showed that the contralateral forelimb could not stretch freely;10%of the neurobehavioral scores were 2 points,showing a tail-fighting sign;5%is 0 points,without any neurobehavioral symptoms;the remai ning 5%are sudden high fever and e.pilepsy.3 Metabolomics study of rat plasma and brain tissue based on UPLC-Q/TOF-MSThe model of focal cerebral ischemia was replicated by filament method.The metabolomics model was established by PCA and OPLS-DA.The results showed that the extraction of lyophilized material from Danshen and Chuanxiong can effectively reverse the model of focal cerebral ischemia and improve the metabolic abnormalities on the focal cerebral ischemia model.4 Metabolomics study of rat striatum based on UPLC-Q/TOF-MSThe focal cerebral ischemia model was replicated by filament method,and the metabolomic analysis method of UPLC-Q/TOF-MS of rat striatum was established by using microdialysis sampling technique.The etabolomics model was established by PCA and OPLS-DA.The differential metabolites identified were(2Z)-7-Hydroxy-2-(phenylmethy-lidene)-heptanoate,Phytosphingosine 3-O-Caffeoylshikimicacid,Leucine,and C16 Sphinganine.ConclusionThe lyophilized of extraction material from Danshen and Chuanxiong has protective effect on cerebral ischemic injury,which can significantly reverse the metabolomics characteristics of plasma and brain tissue disorder,so that the above endogenous substances have been restored to different degrees,It suggests that its protective effect may be related to the regulation of related metabolic pathways.