| Doxorubicin(DOX),a typical anthracycline antineoplastic agent,has limited clinical applications due to its severe and irreversible myocardial damage.In view of its broad-spectrum anticancer effects and high efficacy,exploring effective drugs to reduce its cardiotoxicity is very important.Danhong Injection(DHI)is prepared from two Chinese herbal medicines,Salviae Miltiorrhizae(Danshen)and Flos Carthami(Honghua).DHI has been widely used in clinical to prevent or treat cardio-and cerebro-vascular diseases.It has been proven that DHI could alleviate ischemic myocardial injury,but the effect of DHI against DOX-induced cardiomyopathy has been rarely explored.Therefore,the aim of this dissertation is to systematically investigate the therapeutic effect of DHI on DOX-induced cardiotoxicity(DIC)by chemomics,pharmacodynamics,metabolomics and bioinformatics.The main works of this dissertation are included as follow:1.Research on chemical basis of DHIAn HPLC-ESI-Q-TOF-MS/MS method was established for comprehensive analysis of the chemical constituents in DHI.A total of 52 major compounds were identified or tentatively deduced by comparing their retention time and MS spectra with those of authentic standards or literature data.These compounds could be classified into four categories,including phenolic acids,flavonoids,nucleosides and terpenes.Meanwhile,rat plasma after injected with DHI was analyzed and 29 prototype compounds could be detected in vivo,which may be the important compounds of DHI to exert pharmacodynamic effects.2.Pharmacodynamic studies of DHI against DIC in vitro and in vivoH9c2 myocardial cell model and SD rat model were established respectively to evaluate the protective effect of DHI on DOX-induced myocardial injury.In vitro study showed that DOX could produce obvious cytotoxic effect,decrease cell survival and induced cell apoptosis.While with the intervention of DHI,cell viability significantly increased and cell apoptosis was suppressed.In vivo experiment showed that DHI significantly improved DOX-induced body weight loss and heart weight loss,ameliorated ECG changes,inhibited the elevation of serum biochemical indexes,and increased myocardial antioxidant enzyme activities.The results of in vitro and in vivo studies implied that DHI can effectively alleviate DOX-induced myocardial injury.3.Metabolomics study of the reverse effect of DHI on DICAn LC-MS based non-targeted metabolomics strategy was performed to analyze the metabolic changes in myocardial cells among different groups.31 DIC-related metabolites were detected,which primarily involved in the metabolism of amino acids,purines and pyrimidines,indicating the energy deficiency and oxidative stress induced by DOX.Meanwhile,DHI could restore metabolic disturbances of arginine,GSH,pantothenic acid,cytidine,inosine and 5'-methylthioadenosine.These results suggested that DHI exerted the therapeutic effect against DIC by improving energy metabolism and attenuating oxidative stress4.Bioinformatics-based study on the action mechanisms of DHI against DICThe molecular mechanism of DIC was investigated using gene expression profiles and a total of 1552 differentially expressed genes(DEGs)related to DIC were identified.Functional annotation and enrichment analysis showed that DNA damage,mitochondrial autophagy,mitochondrial translation-related pathways and ErbB signaling pathway were closely associated with the occurrence and development of DIC.Then network pharmacology strategy was employed and 70 potential anti-DIC targets corresponding to 24 active compounds in DHI were identified.KEGG pathway analysis revealed that the cardioprotective effect of DHI against DIC involved in Chemokine,TNF,HIF-1,Toll-like receptor signaling pathways.The active compound-potential target network was further analyzed to obtain key compounds and key targets.The results of RT-PCR suggested that DHI could exert cardioprotective effect against DIC by reversing the expression of PTGS2,ADRB2,CASP3 and RELA. |
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