| Object:1.To develop a reliable qualitative and quantitative method to identify inflammatory mediators and its metabolites.2.To construct the inflammatory mediator profile that captures key features associated with type 2 diabetes and hyperlipidemia.3.To elaborate the mechanism underlying effects of Shenqi Jiangtang Granules intervention in type 2 diabetes(T2DM)and hyperlipidemia based on inflammatory mediator profile.Methods:1.Established method for the analysis of inflammatory mediators87 inflammatory mediators were identified with UPLC-QQQ-MS system and multiple reaction monitoring(MRM)mode,using the characteristic masses of fragment ions for identification and confirmation.Moreover,to establish a quantitative analysis method for inflammatory mediator,the liquid chromatography conditions and mass spectrometry parameters were optimized,and the methodological investigation was perfected.2.Construction of the inflammatory mediator profiles of type 2 diabetes and hyperlipidemiaBased on established inflammatory mediator detection methods,the inflammatory mediator content in plasma samples(including type 2 diabetes and hyperlipidemia rat models)was determined.After standardized processing,univariate and multivariate statistical analyses were used to find the characteristic inflammatory mediators in type 2 diabetes and hyperlipidemia,and established the inflammatory mediator profile of glycolipid metabolic disorders.3.Study on the mechanism of Shenqi Jiangtang Granule in the treatment of glycolipid metabolic disordersThe effects of Shenqi Jiangtang Granule on fasting blood glucose,total cholesterol,triglycerides,low-density lipoprotein cholesterol and high-density lipoprotein cholesterol in type 2 diabetes and hyperlipidemia were observed.Plasma samples of two animal models were analyzed and multivariate data analysis was applied to discover the differential inflammatory mediators and explore the underlying mechanism of Shenqi Jiangtang Granule in the treatment of glycolipid metabolic disorders.Results:1.Established method for the analysis of inflammatory mediatorsAn analytical method for detecting 87 inflammatory mediators in 20 minutes was established by optimization of different parameters.The established method could quickly and accurately quantitatively analyze a variety of inflammatory mediators.All standard calibration curves showed good linear regression,and the correction regression coefficients were between 0.9351 and 0.9999.2.Construction of the inflammatory mediator profile of type 2 diabetes and hyperlipidemiaType 2 diabetes model was constructed by feeding high-fat and highsugar feed and injecting streptozotocin,which had 6 characteristic inflammatory mediators.Hyperlipidemia model was constructed by feeding high-fat and high-sugar feed,which had 10 characteristic inflammatory mediators.The inflammatory mediator profile of glycolipid metabolic disorders contained 13 inflammatory mediators,including(±)12(13)-Di HOME,Eicosapentaenoic Acid,Docosapentaenoic Acid,15-keto-prostaglandins E2,DGLA,Adrenic Acid,(±)9(10)-Di HOME,5(S)-HETE,(±)5-HETE,Arachidonic acid,Linoleic Acid,9(S)-HODE and 13-Oxo ODE.Thirteen inflammatory mediators were involved in the biosynthesis of unsaturated fatty acids,linoleic acid metabolism and arachidonic acid metabolism pathway.3.Study on the mechanism of Shenqi Jiangtang Granule in the treatment of glycolipid metabolic disordersIn the treatment of Shenqi Jiangtang Granule for type 2 diabetes and hyperlipidemia,10 inflammatory mediators had been changed,and all of them were involved in linoleic acid metabolism and arachidonic acid metabolism pathway.Its mechanism of action could be attributed to the following three points,(1)By inhibiting the activity of LOX,blocking the production of 5-HETE and 15-HETE and LTs in the 5-LOX and 15-LOX metabolic pathways,reducing oxidative stress,thereby exerting antiinflammatory effects.(2)By stimulating PPAR-?,inhibiting the activities of CYP2C8 and s EH,increasing the level of EETs in the body,thereby reducing oxidative stress.Conclusions:In this research,an analytical method was established for detecting 87 inflammatory mediators in 20 minutes with simultaneous quantification.The method was successfully applied to the analysis of plasma samples of Shenqi Jiangtang Granule for the treatment of type 2 diabetes and hyperlipidemia animal models,and constructed the inflammatory mediator profile in glycolipid metabolic disorders.It was found that the mechanism of Shenqi Jiangtang Granule in the treatment of glycolipid metabolic disorders was related to 5LOX/15LOX-HETEs and PPAR?-s EH-EETs. |
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