Study On The Chemical Compounds And Monomer Active Of Herpetospermum Seed For The Ethyl Acerate Part

Sijimeiduo,the other name is Herpetospermum seed,which is a common medication used by Tibetan medieinal,it’s the dried ripe seed of Herpetospermum pedunculosum(Sex. C.B.Clarke), it is bitter in taste and cold-natured;It has an effect for purging the fire of liver and gall and detoxifying, The preliminary pharmacological study found that the ethyl acetate fraction with antiviral and liver protection dual role,and it can promote the biliation and have a lipid-lowering effect. The mechanism is through inhibiting the NF-KB inflammatory pathway to achieve the protect effect of anti inflammation. Therefore,it hint us whether it has an anti atherosclerotic effect to impact on cerebrovascular.So, we Separate and purificate the ethyl acetate fraction of the Herpetspermum seed in the study.Then screening the activity of anti cerebral ischemia through using the self constructed HEK293 cell screening model.The last, observing the impact of the picked compound to the effecet on the phenotypic characteristics of rat bone marrow mesenchymal stem cells (rMSCs).Through the two part in the study on the chemical constituents and pharmacological screening it can provide a research foundation for research the efficacy and explore the mechanism of Herpetospermum seed.In the study,we use 95% ethanol to extract the chemical composition of Herpetspermum seed by means of diacolation.Recovering alcohol through the way of vacuum concentration.The concentrated solution dispersed in water, then obtainning three different polar parts through extracting successively with petroleum ether, ethyl acetate and butanol.The sugar and grease in the ethyl acetate extract was removed. Using the technical means include Silica gel column chromatography, Sephadex chromatography, reversed-phase column chromatography, recrystallization to separate and purificate until obtainning the monomer compound.Finally,the compound structure was identified by TLC,1H-NMR,13C-NMR,ect.We identified a total of 9 compounds of the structure:the compounds are Herpetin (BL-1), Herpetene A (BL-2), Eicosanoic,2-propenyl ester(BL-3), Cucurbitacin R (BL-4), Cucurbitacin L(BL-5), 3’-Hydroxydaidzein (BL-6),Dehydrodiconiferyl alcohol (BL-7),Oleanic acid (BL-8),β-Sitosterol (BL-9).The BL-2, BL-3,BL-4, BL-5,BL-6 were first isolated from Herpetospermum seed,and BL-2 is a new compound.The study used 7 compounds to test these cell toxicity,.The activity of the7 compounds was screened by using 2 kinds of cell-based screening models which respectively use VEGF, NGF as target. Through the observation of the effect of the monomer on the expression of red fluorescent protein,The results showed that the monomer BL-2 have an enhanced effect on gene transcriptional activity of NGF, BL-7 have an enhanced effect on the transcriptional activity of VEGF; Through the determination of luciferase activity, the results showed that the monomer BL-2 have an enhanced effect on gene transcriptional activity of NGF, the monomer BL-7 has an enhanced effect on gene transcriptional activity of VEGF. Selecting the monomer BL-2, BL-7 whose the red fluorescent protein and luciferase reporter system are positive to test the dose-effect relationship and the IC50 and EC50, and calculated the IC50 and EC50 values.The study verificated the activity of the compound BL-2 which have an enhanced effect on gene transcriptional activity of NGF by rMSC.Through observing the impact of the compound BL-2 to the effect on the phenotypic characteristics of rat bone marrow mesenchymal stem cells (rMSCs) phenotypes.