Studies On Chemical Constituents And Anti-DN Activity Of Lespedeza Cuneata(Dum.-Cours.) G.Don

This thesis consisted of four chapters.Chapter one studied chemical compositions of ethyl acetate fraction from Lespedeza cuneata（Dum.-Cours.）G.Don.Chapter two described chemical compositions of essential oil extracted from this plant.Chapter three expounded the anti-DN activity of flavonoids from this plant.Chapter four mainly introduced the research status of this plant,including folk medicinal situation,chemical constituents and pharmacological activities.Objective: The objective of this thesis is to study the chemical compositions and anti-DN activity of Lespedeza cuneata.Methods : The compounds were isolated and purified by means of silica column chromatography,MCI and Sephadex LH-20 column chromatography.The structures of compounds were identified by modern spectroscopic techniques（1H-NMR,13C-NMR,2D-NMR）.The essential oil was obtained by steam distillation,and then its chemical compositions were identified by GC-MS.Mesangial cells were randomly divided into three groups: normal control group（glucose concentration of 5.6 m M）,high glucose group（glucose concentration of 30 m M）and medicine group.Each group cells were cultivated 48 hours,and then anti-DN activity in vitro of flavonoids from this plant were detected through the method of WST-1.Results : Thirty-two compounds were obtained from ethyl acetate fraction of Lespedeza cuneata.Based on their spectral data,their structures were identified as2’-hydroxy-4’,6’-dimethoxy chalcone（1）,7-hydroxy-5,8-dimethoxy flavanone（2）,kaempferol（3）,isorhamnetin（4）,quercetin（5）,diosmetin（6）,luteolin（7）,phloretin（8）,lcaritin（9）,cynaroside（10）,rutin（11）,kaempferitrin（12）,lcarrin（13）,aromadendrane-4β,10β-diol（14）,cryptomeridiol（15）,（3R,4R,6S）-3,6-dihydroxy-1-menthene（16）,syringaresinol（17）,isofraxidin（18）,catechol（19）,phenylacetic acid（20）,vanillic acid（21）,4-hydroxycinnamic acid（22）,cyclo（L-Ala-L-Phe）（23）,decanedioic acid（24）,3,4-dihydroxybenzoic acid（25）,cinnamic acid（26）,4-hydroxy-3-methoxy-benzene ethanol（27）,p-hydroxy phenyl ethanol（28）,3,4-dihydroxyphenyl ethanol（29）,hydroxytyrosol acetate（30）,methyl octanoate（31）,β-sitosterol（32）.And compounds1-2,4,8-12,14-20,22-32 were isolated from this plant for the first time.53 compounds of the acrial part were identified,and the main chemical components were phytone（12.44%）,1-Octen-3-ol（6.29%）and heptadecene（5.21%）.19 compounds of the underground part were identified,and the main chemical components were phytol（62.77%）,phthalic acid,isobutyl octyl ester（3.83%）,methyl hexadecanoate（3.47%）.There was significant difference between the two parts,and the acrial part contained more components than underground part.The results showed that high glucose clearly induced the proliferation of MC.The method of WST-1 was used to detect cell proliferalion.Compounds 1,3,6,7,8,9,10,12,13 inhibited the proliferation of MC,which was induced by high glucose.There was dose dependence of compounds 1,3,6,7,10,12,13 in a certain concentration range.Based on the calculation of the IC₅₀,we found that compound 7 had obvious anti-DN activity.Conclusion:All compounds were isolated from this plant for the first time,except compounds 3,5-7,13 and 21.The variety and content of essential oil extracted from acrial and underground parts had great difference.There was dose dependence of compounds 6,7,10 in a certain concentration range.The compound 7 had obviously effect.