Study On The Components Of Diterpenoids And Their Regulation Of Glucose Metabolism

Objective:The research object of this paper is the whole plant of the Scoparia dulcis L.The diterpenoids in this plant was separated by various separation techniques and its hypoglycemic activity was studied.The effect of diterpenoid compounds on the glucose consumption of Hep G2 cells was studied by glucose oxidase method,the effect of diterpenoids on the activity of MIN-6 cells after injury induced by palmitate acid(PA)was detectd by MTT method,and based on network pharmacology,the antidiabetic mechanism of diterpenoids from Scoparia dulcis L was studied.Through the above study,we could provide reference for the effect of diterpene of Scoparia dulcis L on the glucose consumption of Hep G2 cells,the cell viability of MIN-6 cells after apoptosis and the may target protein of diterpene of Scoparia dulcis L acting on diabetes.Methods:1、The whole plant of Scoparia dulcis L was refluxed and extracted with 85% ethanol aqueous solution.The extract was concentrated to be alcohol free under reduced pressure.After the extract was dissolved with water,the fat soluble components were removed by petroleum ether extraction.Then the mother liquor was extracted with dichloromethane,the organic phase in the lower layer was recovered,and the dichloromethane was evaporated by heating under reduced pressure to obtain the dichloromethane extract.Dichloromethane extract was separated by chromatography,such as silica gel column chromatography(CC),gel column chromatography(Sephadex LH-20)and liquid chromatography(HPLC),etc.The structure of the isolated compounds was identified according to the spectral data(HR-ESIMS、1D NMR、2D NMR),etc.2、Glucose oxidase method was used to detect the effect of diterpenoids of Scoparia dulcis L on glucose consumption of Hep G2 cells.3、MTT method was used to detect the effect of diterpenoids of Scoparia dulcis L on the viability of MIN-6 cells after injury induced by PA.4、Based on the network pharmacology,study the mechanism of the anti-diabetes action of the diterpenoids of Scoparia dulcis L.Results:1、12 diterpenoids were isolated from the whole plant of Scoparia dulcis L,identified separately as Scoparicol A(1)、Scoparicol B(2)、Scopadulcic acid D(3)、Scoparic acid E(4)、Scopadulcic acid E(5)、Iso-dulcinol(6)、Scopadulcic acid C(7)、Dulcidiol(8)、Scopadulciol/Dulcinol(9)、Scoparic acid B(10)、Scopadulcic acid B(11)、Scoparic acid C(12).Five of these compounds(1-5)are new compounds not reported in the literature.2、The effects of compounds 6,7,9,10,11 on glucose consumption of Hep G2 cells were studied.The results showed that compound 7 could significantly increase the glucose consumption of Hep G2 cells.3、In this paper,the effects of compounds 4,6,7,9,10,11 on the cell viability against MIN-6 cells were tested.The results showed that,comparesd with cell viability of model group(34.48±0.82)%,compouns 3,6,7 and 10 could increase the activity of MIN-6 cells at concentration of 25 μmol/L,the cell viabilities were(41.31±3.64)%,(40.17±3.48)%,(40.64±2.92)%,(39.79±6.78)% respectively,compounds 7 and 10 could significantly increase the viability of MIN-6 cells at concentration of 50μmol/L,the cell viabilities were(45.74±1.50)% and(42.47±2.60)% respectively.The other compounds did not increase the viability of MIN-6 cells.4、 The results showed that PIK3 CA,TNF,MAPK1 and TP53 are the core target proteins of diterpenoids of Scoparia dulcis L acting on diabetes.Conclusions:1、12 diterpenoids were found,including 5 new compounds.2、Compound 7 can promote the sugar consumption of(HepG2)cells.3、Compounds 3,6,7 and 10 can increase the viability of MIN-6 cells after injury induced by PA at concentration of 25 μmol/L;Compounds 7 and 10 can significantly increase the viability of MIN-6 cells after injury induced by PA at concentration of 50μmol/L4、PIK3CA,TNF,MAPK1 and TP53 are the core target proteins of diterpenoids from Scoparia dulcis L acting on diabetes.