Metabolomics Study Of The Decocted Solution From Shuang-Dan Preparation On Anti-acute Myocardial Ischemia Effect

Shuang-Dan preparations,a compatibility composition of two Traditional Chinese Medicines including Radix Salviae Miltiorrhizae and Cortex Moutan,were used in the treatment of blood stasis and activating meridians.Modern pharmacological studies showed that Shuang-Dan preparations played an important role in the treatment of many cardiovascular diseases.However,its therapeutic mechanism was still unknown.In present thesis,the decocted solution from Shuang-Dan preparation(Shuang-Dan decoction),prepared by the aqueous extraction of Radix Salviae Miltiorrhizae and Cortex Moutan(2:1),was investigated the changes of chemical composition in vitro and metabolite response in vivo by HPLC-MS based on metabolomics.(1)HPLC-MS technique was applied for analyzing Shuang-Dan decoction to reveal changes of chemical components after the compatibility of Salvia Miltiorrhiza and Cortex Moutan.The results indicated that some phenolic acids such as Danshensu,Caffeic acid,Rosmarinic acid,Protocatechuic acid,Salvianolic acid A,Salvianolic acid B,Salvianolic acid C,Salvianolic acid D,Salvianolic acid E,Neochlorogenic acid and Lithospermic acid,are derived from Radix Salviae Miltiorrhizae.In addition,some compounds such as Myoinositol,Sucrose,Citric acid,Gallic acid,Oxypaeoniflorin,Mudanoside B,Suffruticoside D,Mudanpioside E,Paeoniflorin,Galloylpaeoniflorin,Iso-mudanpioside H,Benzoyloxypaeoniflorin,Mudanpioside J are derived from Cortex Moutan.Most importantly,a novel compound,tentatively named Lamiide,was discovered in Shuang-Dan decoction.Lamiide was widely distributed in the Labiatae plant and had anti-oxidation and free radical scavenging effects.Whether the substance played a key role for treating myocardial ischemia in Shuangdan preparation needed to be studied further.(2)Anti-myocardial ischemia model in rat was induced by subcutaneous injection of isoproterenol(100 mg/kg)and this pathological model was validated by typical plasma clinical biochemical indexes including LDH,CK,CK-MB and AST.HPLC-MS based metabolomic method was applied to explore anti-myocardial ischemia effect of Shuang-Dan decoction.Compared with the normal group,54 metabolites in plasma,38 metabolites in urine,23 metabolites in fecal samples and 44 metabolites in four tissures including heart,liver,brain and kidney were significantly changed.These extracts were adjusted to some degrees after offering Shuang-Dan decoction,indicating that these metabolites were closely related to the effect of Shuang Dan on acute myocardial ischemia and involved with amino acid metabolism,neurotransmitter metabolism,energy metabolism,fatty acid metabolism and carbohydrate metabolism.It was suggested that the metabolic pathways involved with amino acid metabolism,neurotransmitter metabolism,energy metabolism,fatty acid metabolism and carbohydrate metabolism.These results could be helpful to interpret the anti-myocardial ischemia mechanism of Shuang-Dan decoction,and enhance the understanding of the combination of Traditional Chinese Medinces.