Study On The Chemical Compouds Of Huang Lian Jie Du Decoction And Metabolomics Study On The Protective Effect Of Diabetes Encephalopathy

Huang Lian Jie Du Decoction(HLJDD)is a traditional heat-clearing and detoxifying Chinese medicine compound,which was firstly recorded in Handbook of Prescriptions for Emergency.It was attending the heat burn and the tripe energizer excessive heat syndrome,which was comprised of four herbs,including Coptidis Rhizoma,Scutellar-iae Radix,Phellodendri Cortex,and Gardeniae Fructus with a ratio of 3:2:2:3.In recent years,the chemical components of HLJDD,has been intensively studies.However,the research focused on one or more of its components.Therefore,it is necessary to estabolish a reliable analytical method for the chemical constituents of HLJDD.Modern pharmacological studies have shown that HLJDD has a therapeutic effect on diabetes encephalopathy(DE).However,the pharmacodynamic substances and the mechanism are not clear.Therefore,aiming at these problems above,the studies showed in vivo and in vitro chemical constituents of HLJDD,and the pharmacodynamic substances and the mechanism of HLJDD to treat diabetes encephalopathy by metabolomics.The specific research contents and results are as follows: 1.Analysis of chemical constituents of HLJDDIn the study,the chemical constituents of HLJDD separated were identified by ulta-perfomance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometer(UPLC-Q Exactive-Orbitrap MS)combined with the compound data of HLJDD.Firstly,it was established HLJDD compound database,which include the information of chemical constituents from the target Chinese traditional medicine base on the literature.138 chemical compounds,including 39 alkaloids,29 flavonoids,15 iridoids,5 monoterpene,36 organic acids and 12 other types constituents,were identified by the methods,which including the fragmentation patterns for diffenrent types of known constituents,Compound Discover software,the related literatures,and the reference substances.It provides data support for the analysis of components of HLJDD in vivo.2.Analysis of prototype components of HLJDD in vivoBase on the chemical components of HLJDD,UPLC-Q Exactive-Orbitrap MS was used to analyze the prototype components in vivo of HLJDD,which distribution in various tissues and organs.By,The chromatographic peaks of the prototype components were determined by comparing the biological samples of the rats that given the drug with the mass spectrometry information of the HLJDD extract and subtracting the matrix peaks from blank biological samples.As a result,31 prototype components were characterized in the drug plasma for the first time,which including 12 alkaloids,13 flavonoids,and 6 iridoids.This study was shown that alkaloids were mainly distributed in kidney,liver and brain,flavonoids were mainly distributed in liver and heart and iridoids were distributed in all tissues.3.Pharmacodynamic study on the protective effects HLJDD on DEDiabetes encephalopathy mice model was established by db/db mice.The study was shown that HLJDD water extract could protect the neuronal cells and improve the learning capacity and the memory function in diabetes encephalopathy mice by Morris water maze experiment,pathological sections,and biaochemical index,which including SOD,GSH,Ach,and MDA level.4.Metabonomics study on the protective effects HLJDD on DE In the current study,an UPLC-Q Exactive-Orbitrap MS method was applied to study the plasma metabolites of HLJDD to treat DE.The multivariate statistical analysis had showed that there were significant differences in plasma metabolites between the administration group and the model group as well as between the model group and the blank group.As a result,it was identified 21 potential biomarkers,including unsaturated fatty acids,and lysophosphatidylcholine,which were involved in the oxidation of long chain fatty acids,linoleic acid metabolism and glutathione metabolism.Therefore,HLJDD play a role in the protective effects on DE,such as neurotransmitter metabolism,regulation of lipid metabolism disorders,antioxidant damage,and promoting anti-inflammatory reactions.In conclusion,it established a method for vivo and vitro analysis of HLJDD constituents,which the chemical composition in vitro and in vivo of HLJDD.The result illustrated that different types of components of HLJDD specificity distribute various tissues and organs,and preliminary illustrates HLJDD has a therapeutic effect on diabetes encephalopathy by regulating lipid metabolism,anti-oxidation.