Studies On The Chemical Constituents From The Roots Of Euscaphis Japonica And Their Inhibitory Activity On Triglyceride Accumulation

Euscaphis japonica is one of species of the genus Euscaphis,which belongs to the family Staphyleaceae.It possesses general pharmacological activities and its root,flowers,nuts are available for medicinal purposes.Previous pharmacological research reported that the methanol extract of E.japonica possesses anti-inflammatory,anti-hepatic fibrotic activity and downregulating lip genesis properties.To better take advantage of the medicinal plants,the aim of this work was to study on the chemical constituents from the roots of Euscaphis japonica and their inhibitory activities on triglyceride accumulation.Various chromatography technique such as column chromatography on silica gel,MPLC and preparative HPLC were used to isolate and purify the compounds and their structures were identified by the spectral data and physicochemical properties.They were identified as(12R,13S)-3-methoxy-12,13-cyclo-taraxerene-2,14-diene-1-one-28-oic-acid(1),3,7-dihydroxy-5-octanolide(2),Bergapten(3),β-sitosterols(4),7-Hydroxy-2-octen-5-olide(5),3,3’-methoxy-ellagicacid(6),4-hydroxy-3-methoxybenzaldehyde(7),Gallic acid(8).Among them,compound 1 was a new compound and compound 8 was isolated from the genus Euscaphis for the first time.In order to explore inhibitory activity on triglyceride accumulation of Euscaphis japonica,in this research,HepG2 cells were treated with oleate in the simulation of liver lipid accumulation and hepatic steatosis.The effect of different polar fractions from Euscaphis japonica ethanol extract and compound 1 on lipid accumulation were studied.Their effects tested by the Oil Red O experimental,the quantitative determination of triglycerides kit experimental and their cytotoxicity on HepG2 cells by MTT assay.The results indicated that:the total ethanol extract from Euscaphis japonica shows no obvious toxicity in dose no more than 100μg/mL but shows a certain inhibiting lipid accumulation effect.In the dose no more than 50 μg/mL,four different polarities extract shows no obvious toxicity.Petroleum ether,ethyl acetate extract significantly inhibited the accumulation of HepG2 cells and reduce the amount of triglyceride,but n-butyl alcohol and water extract do not show significant inhibitive activity.New Compound 1 did not show obvious cytotoxicity to HepG2 cells up to 25μg/mL but did affect cell viability at the concentration of 50 μg/mL.It was inhibition fat accumulation and reduce the content of intracellular triglyceride in dose-dependent manner.Therefore,the alcohol extract of Euscaphis root shows significantly inhibitory activity of lipid accumulation,where petroleum ether fraction(in the low polar)and ethyl acetate fraction(middle parts)are its main active fraction.Compound 1 is the important active ingredient of Euscaphis root.As an important activity of inhibitory lipid accumulation,compound 1 is expected to development of lead compound.