Study On The Chemical Components,Quality Evaluation And Antitumor Activity Of Scutellaria Barbata D.Don

Scutellaria barbata D.Don was the subject of the study and a series of research about its chemical composition,quality control and mechanisms were developed.The doctoral thesis is divided into five distinct parts which are reviews,resear of chemical compositions,quality control,activities and mechanisms in vivo and in vitro and the interaction relationship of dug effect superposition and relation of inflammation to tumour.The purpose was to preliminarily study the therapeutic material basis and the relation of inflammation to tumour of Scutellaria barbata D.Don by technical means of isolation and identification of chenical constituents,quality control,metabonomics,etc.Chapter 1.Literature researchThis part was reviewed on quality control,chemical composition and mechanisms of Scutellaria barbata D.Don by organizing literature.The background and research directions of doctorial thesis were also expounded systematically.Chapter2.Research on chemical compositionsChemical compositions of Scutellaria barbata D.Don were separateed and identified by extracting,silica gel,Sephadex Gel,C18 column and MCI,MPLC,HPLC,UPLC,UV identification,NMR and reference standards.26 chemical compositions were separateed and identified and 13 of them were flavonoids,which were the chemical basis for further research.Chapter3.Quality control of Scutellaria barbata D.DonThe Chinese pharmacopoeia(Ch.P.)of the 2015 edition about Scutellaria barbata D.Don was revised.The moisture content,total ash,acid-insoluble ash,heavy metal concentrations,extract content,total flavonoids content,trait identification,microscopic identification,scutellarin and luteolin content were checked or tested to provide basis on quality control of Scutellaria barbata D.Don.In addition,characteristic fingerprint of Scutellaria barbata D.Don from 18 different producing areas and batches was established by HPLC and LC-MS.14 common peaks were identified and 19 chemical constituents were identificated by LC-MS and reference standards.Chapter4.The activity of in Vivo and Vitro of Scutellaria barbata D.DonWater extractive fractions and ethanol extractive fractions were investigated by MTT method first and ethanol extractive fractions were choosed as the follow-up study subject.Then comparison in different extractive fractions was did and the active constituent of Scutellaria barbata D.Don may distribute in dichloromethane extraction or ethyl acetate extraction.We also found that the total flavonoids of Scutellaria barbata D.Don induced SMMC-7721 cell apoptosis and there's a connection between IL-6/STAT3 and NF-kB signaling pathways in the follow-up research.Antitumor research in vivo about the total flavonoids of Scutellaria barbata D.Don was conducted by model of mouse hepatoma H22 and it showed activity of antitumor in vivo.It proved that the total flavonoids may down regulation of IL-6/STAT3 and NF-kB signaling pathways to inhibit tumor by methods of ELASA,immunohistochemical analysis,Western Blot,etc.The blood composition was also researched to test the effectiveness of total flavonoids by methods of MTT and TIC of LC-MS At last,hispidulin was used as the representative chemical composition to test the variety of the metabolites in H22 ascites tumorin mices and confirmed that they had links to TCA cycle.Chapter5.Investigation of the relationship between inflammation and tumour on Scutellaria barbata D.Don as well as the theory of dug effect superposition based on metabonomics of cellScutellaria barbata D.Don may accord with the theory of dug effect superposition by the results of preliminary experiments.Medical materials from two different producing areas were screened by a series of statistical methods MTT of them were also tested and found that results were closely aligned,which validated the theory initially.Metabonomics research of S14 and S17 were applied by SMMC-7721,LO2,RAW264.7 and their models induced by LPS.We found that their distribution maps of OPLSD and metabolic pathways were close to unanimous,which further validated the theory of dug effect superposition.The main metabolic pathways of S14 and S17 are lycerophosphatide,arachidonic acid metabolism,riboflavin metabolism and Krebs cycle,which have a close relation to IL-6/STAT3 and NF-kB signaling pathways.It also validated the mechanisms in Chapter 4.