Studies On Chemical Constituents From The N-butanol Extract Of Potentilla Anserine L And The Anti-hypoxia Activity

Potentilla anserina L.is a medicinal herb of genus Potentilla in the family Rosaceae,which is widely distributed in the western areas of China,especially in Tibet,Gansu and Qinghai provinces.For thousands of years,this Tibetan tradiotonal medicine has been popularly used for its curative properties.Our previous pharmacological studies(in vitro and in vivo) indicted the n-butanol extract of P. anserine L.showed a remarkably protective effect on cardiac myocyte during hypoxia by improving the ability of cleaning oxygen free radial in cardiac myocyte and inhibiting the oxide injury to membrane.Therefore,it is supposed to be the bioactive fraction of P.anserine L.However,little has ever been known about the phytochemical profile of the n-butanol P.anserine L.fraction and the bioactive components,which can explain the anti-hypoxia effect,are still unknown.Addtionally, a phytochemical study is of high value in order to confirm anti-hypoxia activity in the light of a rational phytotherapy.In the present study,the bioguided-fractionation of 70%ethanol extract of P. anserina L.was carried out.19 compounds were isolated from the n-butanol extract by open column chromatography such as silica gel,ODS and Sephadex LH-20,PTLC and preparative RP-HPLC.On the basis of physico-chemical properties and spectroscopic analysis(ESI-MS,¹H-NMR,¹³C-NMR and 2D-NMR),14 compounds were structurally elucidated asβ-sitosterol(1),ursolic acid(2),euscaphic acid(3), tormentic acid(4),rosamultin(5),kaji-ichigoside F₁(6),24-deoxy-sericoside(7), 2-oxo-pomolic acid-β-D-glucopyranosyl ester(8),β-daucostarin(9),adenosine(10), daidzin(11),puerarin(12),3'-methoxypuerarin(13),daidzein 8-C-apiosyl(1→6) glucoside(14).Among them,Compound 11,12,13 and 14 were isolated from genus Potentilla for the first time.Compound 10 was first isolated from this plant.Meanwhile,by the hypobaric hypoxia in mice,the brain water content,the activity of SOD and the content of MDA in brain and serum were determined.The result suggested the n-butanol extract of P.anserine L.had the protective effect on brain injury induced by hypobaric hypoxia.According to the previous reports,we conclude that the adenosine(10) and four isoflavone compounds(11～14) may partly be responsible for its anti-hypoxia ability.In conclusion,this research has provided rational and experimental basis for further development of the plant.