Objective: To systematically screen the hepatoprotective compounds from ethyl acetate extract of the roots of Indigofera stachyoides. Method: The roots of I. stachyoides were extracted by ethanol

and then its extract was further divided into ethyl acetate fraction and n-butanol fraction. The hepatoprotective activity screening was performed in mice in the normal control group

carbon tetrachloride (CCl4) model group

bifendate (DDB) group

intragastric administration group and subcutaneous group of ethyl acetate fraction

intragastric administration group and subcutaneous group of the n-butanol fraction. Glutamic pyruvic transaminase (ALT) level was detected in miceto evaluate the hepatoprotective activity of different samples. Then effective compounds of the hepatoprotective sample had been isolated and their structures were identified by silica gel column chromatography

Sephadex LH-20

RP-18 column chromatography and other means. Screening of hepatoprotective activity of isolated chemical compounds was performed in vitro in CCl4-induced human HL-7702 cell damage model. Result: Subcutaneous group of ethyl acetate fraction showed significant hepatoprotective effect on mice liver injury model (P<0.05). Total 11 compounds were isolated from this ethyl acetate fraction of I. stachyoides

which had been identified as stigmasterol (compound 1 )

L-maackiain (compound 2 )

medicarpin (compound 3 )

stigmast-4-en-3-one (compound 4 )

stigmastan-3-one (compound 5 )

3-hydroxy-8

9-dimethoxypterocarpan(compound 6 )

(+)-epicatechin(compound 7 )

2α

3α-epoxyflavan-5

7

3'

4'-tetraol-(4β→8)-flavan-5″

7″

-4'"-triol (compound 8 )

2α

3α-epoxy-5

7

3'

4'-tetrahydroxyflavan-(4β→8)-epicatechin (compound 9 )

(-)-epicatechin (compound 10 ) and schizandriside (compound 11 ). Compound 2 was isolated from this plant for the first time. Compounds 3 and 6 were isolated from Indigofera for the first time. Compounds 8

9 and 11 had a certain degree of protective effect on CCl4-induced human HL-7702 cell damage model (P<0.05). The survival rate of HL-7702 was significantly increased

suggesting that compound 8

compound 9 and compound 11 had protective effects on hepatocyte injury in vitro. Conclusion: In this study

ethyl acetate fraction of I. stachyoides was the main effective part of the hepatoprotective effect

and compound 8

9 and 11 from this fraction were the main effective compounds.