The Pharmacological Components And Mechanism Of Salvia Miltiorrhiza Were Found Based On The Myocardial Injury Model Induced By Doxorubicin

The efficacy of chemotherapeutic agents has improved prognosis for cancer patients,yet chemotherapy-induced cardiomyopathy remains an important adverse outcome of treatment.There are no consensus guidelines regarding prevention,monitoring,or treatment of patients at risk for chemotherapy-induced cardiomyopathy,and patients with cancer are often excluded from large heart failure trials.Adriamycin(Doxorubicin,DOX)is a potent and widely used anthracycline antibiotic for the treatment of cancers.However,Adriamycin caused irreversible myocardial damage,eventually leading to heart failure in clinic.The mechanism of Adriamycin-induced myocardial damaged relatively complex.Recent studies suggest that the main mechanisms of Adriamycin-induced myocardial damaged included mitochondrial damage,oxidative stress,calcium overload and cytogenetic toxicity.As most biological processes,the myocardial damage is dynamic and related to time-series.Time-series gene expression data analysis provides the information dependent on time for the study of biological processes,including the differences gene expression in static state and dynamic relationship of them.Though the time-series gene expression data analysis,the biological network of disease progression can be built in more detail and help for drug development and clinical treatment.In this study,we use Salvia miltiorrhiza to find out the effect fraction and components against Adriamycin-induced myocardial damaged,and study those mechanism in protecting myocardium.Four major study contents are as follows:1.Time-series gene expression data analysis of adriamycin-induced rat myocardium injuryThe transcriptome arrays were used to detect adriamycin-induced rat myocardium injury which download from DrugMatrix database.At first,the GC robust multi-array average were used in background adjust.Difference expression genes were detected in different observation time points which used the normal group as control.Cutoff value are 1.5-fold and p-value were less than 0.05.Difference expression genes in three time points were 815 in total.Next,Short Time-series Expression Miner toolbox were used to analysis the gene expression profile.There were 9 different gene expression profile.Each gene group were analyzed by GO to figure out the main effect genes which include 196 genes.STRING as a tool to search the protein-protein interaction of 196 genes.The dynamic biological network of adriamycin-induced rat myocardium injury was built by 196 genes and 563 nodes which provide the basis of mechanism study.2.The model of adriamycin-injured rat H9C2 cardiomyocytesThe cell model of adriamycin-injured rat H9C2 cardiomyocytes were established by high content real time cell fluorescence technique based on dynamic biological network,the results of part 1.Several parameters which related the mitochondrial damage,calcium overload and cytogenetic toxicity were acquired by this technique.Edaravone,free radical scavenger,used as a positive control.Results show that edaravone in 10?g,M inhibited cell apoptosis and calcium overload induced by adriamycin,and had no effects on cytogenetic toxicity.All results indicated that the model could be used for screening.3.Preparation of Salvia miltiorrhiza fraction and quantitative analysisThe method for quantitative analysis of salvia miltiorrhiza by HPLC were developed,include danshensu,protocatechuic aldehyde,ferulic acid,acacia,salvianolic acid D,salvianolic acid B,salvianolic acid A,salvianolic acid C,carnosol,dihydrometone I,cryptotanshinone,tanshinone,tanshinone IIA,oleanolic acid.By using liquid chromatography method transform,the analysis method convert to micro-preparation method.In results,six fractions of salvia miltiorrhiza were prepared and quantitative analysis.The molar concentration of Salvianolic acid B is 4.02 mmol per liter in fraction.4.The compounds screening of salvia miltiorrhiza and mechanism studyThe cytotoxicity of salvia miltiorrhiza fractions were measured by H9C2 cells,then all fractions were used to evaluate myocardial protection by the cell model which established in part 2.Results show that fraction 1 and fraction 2 inhibited Adriamycin-induced myocardial damaged.76 compounds in TCM library and 14 compounds from salvia miltiorrhiza were screened by the cell model.The results were analyzed by PCA-K-means method.The protective effects of Danshensu,protocatechuic aldehyde,ferulic acid,acacia,salvianolic acid A,dihydro tanshinone were same as the effects of edaravone.The hits from salvia miltiorrhiza were from fraction 1 and 2 which as the results in fractions active evaluation assay.In the study of protection mechanism,we search the targets of those compounds by TCMD database.MMP2?ALOX15?APP?CYP2E1 were match in compounds targets and dynamic biological network which established in part 1.