Chemical Composition Of Agrimonia Pilosa Ledeb. And Its Hypoglycemic Activity

Xian he cao(Agrimony)is the dry aerial part of Agrimonia pilosa Ledeb.It belongs to Rosaceae family and has the functions of astringent and hemostatic,antimalarial,antidysentery and detoxification.It is regarded as good folk medicine for diabetes with a wide application in clinical treatment of diabetes and reducing patients’ blood sugar.Triterpenoids in Agrimonia pilosa have been reported to have certain hypoglycemic activity.However,further research is needed for the affects of material basis and mechanism of the Agrimonia pilosa hypoglycemic.Therefore,Agrimonia pilosa was chosen to be the research object in this paper to find compounds with hypoglycemic activity and clarify its mechanism of action.Agrimonia pilosa was extracted with 60%ethanol by heating and refluxing.Through Sephadex LH-20 gel column chromatography,silica gel column chromatography,reversed-phase column chromatography,semi-preparative HPLC,and recrystallization,28 compounds were isolated and purified from the ethanol extract of Agrimonia pilosa.The HR-ESI-MS,NMR spectroscopy,ultraviolet absorption spectroscopy,and infrared absorption spectroscopy were used to analyze the plane structure of the compound,and the absolute configuration of the compound was determined by combining circular dichroism spectrum and specific rotation.Among them,compounds 1,2,15 are new compounds,compounds 1-14 are triterpenoids,compounds 15、16 are phloroglucinol derivatives,compounds 17-22 are organic acid compounds and compounds 23-28 are other types of compounds.Further research on the inhibitory effects of Agrimony’s triterpenoids onα-glucosidase,the results showed that compounds 3-5 had a significant inhibitory effect on α-glucosidase(IC50=18.9,23.3,27.1 μmol/L),which were stronger than the positive drug(Acarbose),compound 8-10 also has a certain inhibitory activity.The experimental results of α-glucosidase inhibition kinetics shows that the inhibitory effects of compounds 3-5 on α-glucosidase belongs to reversible inhibition,and its inhibition type belongs to anti-competitive inhibition.Further confirmation through fluorescence quenching,CD and molecular docking that compound 3-5 has a direct interaction with α-glucosidase,which can cause changes in secondary structure.PDX-1 plays an important role in Pancreatic β-cell differentiation and mature Pancreatic β-cell maintenance.Our study further investigated the effect of Agrimolide(compound 25)on the expression of PDX-1,and the results showed that 5 μmol/L compound 25 promoted the expression of PDX-1.Our research has initially revealed the material basis and mechanism of the hypoglycemic activity of Agrimony,which provides a reference for the further rational development and utilization of Agrimony herb materials.