Metabolomics Study On Anti-inflammatory Effects Of Monomeric Compounds In Salvia Miltiorrhiza Bge. Based On UPLC Q-TOF/MS

To shed light on the properties of polar metabolome in inflammatory cells,we selected LPS induced RAW264.7 inflammatory cell model as the carrier for the research of metabolic fingerprint analysis.In this study,a UPLC Q-TOF-MS-based metabolomics protocol was optimized for the extraction of polar metabolites from RAW264.7 cell line.Orthogonal partial least squares discriminant analysis(OPLS-DA)was used to process the metabolic data,a total of 17 metabolites were selected and identified.The results showed that Methanol/Chloroform/H2O(8:1:1,v/v/v)was chosen as the optimal extraction solvent to recover higher number of intracellular features with the best relative extraction efficiency and stability.Comparing with the normal cells,the inflammatory cells present an abnormal metabolism in protein,carbohydrate,nucleotide and phospholipids.In this study,a UPLC Q-TOF-MS-based metabolomics protocol for the polar metabolites from RAW264.7 cell line was developed,which may provide important information for the study of mechanism of inflammation and the anti-inflammatory drugs.Salvia miltiorrhiza has a effect of enhancing immune and anti-inflammatory.The study selected compounds in Salvia(tanshinone IIA,cryptotanshinone,tanshinol and salvianolic acid A)as the object to metabonomics research of Salvia miltiorrhiza monomer compounds that interfere inflammatory cell model,aiming at revealing their anti-inflammatory effects mechanism from cellular metabolism level.In order to study compounds in Salvia on RAW264.7 cells induced by LPS the anti-inflammatory mechanisms of inflammation,carried out the following research:1.Filter RAW264.7 cells induced by LPS model number of cells;2.Select MTT method combining Griess experiment establish dose of compounds in Salvia;3.Do the metabonomics studies of Salvia miltiorrhiza monomer compounds that interfere with the inflammatory cell model.The intracellular metabolite of normal groups,model groups,administration of tanshinone IIA groups,cryptotanshinone groups,tanshinol groups and salvianolic acid A groups were detected by UPLC Q-TOF/MS.Orthogonal partial least squares discriminant analysis(OPLS-DA)was used to process the metabolic data,normal groups,model groups were completely separated and administration of tanshinone IIA groups,cryptotanshinone groups,tanshinol groups and salvianolic acid A groups close to the control group.The results show that the inflammation model was successful established.Tanshinone IIA,cryptotanshinone,tanshinol and salvianolic acid A make a difference on RAW264.7 cells inflammatory model induced by LPS,and show a return to normal trend.Results show that monomer compounds in Salvia miltiorrhiza can callback the changed metabolites in inflammation,by adjusting the citric acid cycle,improving glucose and fatty acid metabolism disorders recover to achieve anti-inflammatory effects.