| Objective:to study the chemical components of the ethyl acetate of Indigofera stachyoides Lindl.and to screen the isolated compounds for their hepatoprotectiveactivityinvitro.Finally,liverprotectivepropertiesof2?,3?-epoxy-5,7,3?,4?-tetrahydroxyflavan-?4??8?-epicatechin?11?with liver protective properties were studied.Methods:1.Silica column chromatography,MCI column,Sephadex LH-20,RP-18,HPLC and other different methods and technologies were used to separate the active monomer compounds.2.Paracetamol?APAP?with the optimal injury concentration and time was used to induce human normal liver cells?HL-7702 cells?to establish an in vitro injury model of liver cells,and the isolated compounds were screened for liver protective activity in vitro.HL-7702 cells at logarithmic growth stage were inoculated into 6-well plates at a concentration of 1×105/mL,2 mL per well.The experiment was divided into three groups,the blank control group.APAP injury group;Drug protection group?40?mol/L?80?mol/L?160?mol/L?.It was cultured in an incubator at 37oC and 5%CO2for 24 h.After being cell adherent completely replace the blank group and model group completely RPMI 1640 culture,drug protection set of replacement for the corresponding concentration of nutrient solution,medicated 12 h after continue to develop,in addition to the control group,the rest of the group to join the best damage APAP solution concentration,after 12 h,respectively collect groups of cells that used to detect supernatant fluid reducing glutathione?GSH?,alanine aminotransferase?ALT?,aspertate aminotransferase?AST?,malondialdehyde?MDA?,and other indicators,research of compound 11 protect liver function.Results:1.Indigofera stachyoides Lindl.ethyl acetate part were separated and identified 11 compounds,?-sitosterol-D-glucoside?1?,schizandriside?2?,kaempferol-7-O-?-D-glucopyranoside?3?,stigmasterol?4?,3,4,5-trimethoxyphenyl-O-?-D-Glucopyranoside?5?,2-methoxy-4-?2'-hydroxyethyl?-phenol-1-O-?-D-glucopyranoside?6?,2-?3-hydroxy-4-methoxyphenyl?ethyl1-O-?-D-glucopyranoside?7?,maackiain?8?,7-hydroxy-2-?4'-methoxyphenyl?chroman-4-one?9?,calycosin?10?,2?,3?-epoxy-5,7,3?,4?-tetrahydroxyflavan-?4??8?-epicatechin?11?the structure identification of certain:.Compounds 5,6,7 and 9 were isolated from leguminous plants for the first time.Compounds 3 and 10 were isolated from cyanobacteria for the first time.Compound 1 was isolated from Indigofera stachyoides Lindl.for the first time.2.After activity screening,compounds 2,9,10 and 11 were found to have a protective effect on apap-induced HL-7702 cell injury in human liver,and the cell survival rate was significantly increased.3.Compound 11 can reduce the activity of ALT and AST,decrease the content of MDA and increase the content of GSH in the HL-7702 hepatocyte injury model.Conclusion:1.this study further enriched and improved the chemical composition of Indigofera stachyoides Lindl.on the basis of the existing research.2.Compounds 2,9,10,11 have protective effects on HL-7702injury induced by paracetamol in human hepatocytes in vitro,among which compounds 10 and 11 have better hepatoprotective activity.3.compound 11 has a protective effect on apap-induced human liver HL-7702 cells.Its protective effect may be through reducing the activity of ALT and AST in cells,and increasing the content of MDA and GSH,so as to exert its hepatoprotective activity. |
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