Activity Constituents Of Malus Halliana Koehne And Ligustrum Lucidum Ait

This paper is composed of three chapters.In first chapter,chemical composition and anticoagulant effect of Malus halliana Koehne flowers were investigated.In second chapter,the dynamic changes in its secondary metabolites during blooming were investigated.The changes in the contents of three flavonoids(phloretin-4'-O-glycosidase,afzeloside,and 3-hydroxyphloridzin)were determined by high performance liquid chromatography(HPLC).In third chapter,chemical composition and glucose uptake effect on 3T3-L1 adipocytes of Ligustrum lucidum Ait.flowers were investigated.Chapter 1 Chemical composition and anticoagulant effect of Malus halliana Koehne flowersMalus halliana Koehne flowers have been used as a traditional Chinese Medicine to treat metrorrhagia,its chemical composition and anticoagulant effect were investigated.Seven compounds were isolated and identified from M.halliana flowers,including limocitrin-3-O-glucoside(1),baohuoside ?(2),kaempferol-3-O-?-L-furan arabinoside(3),phloretin-4'-O-glycosidase(4)and afzeloside(5),phloridzin(6),3-hydroxyphloridzin(7).All compounds were isolated for the first time from M.halliana flowers,compound 1-4 were isolated for the first time from M.halliana and compounds 1-3 were isolated for the first time from in Malus.Then the anticoagulant effect of the compounds and extracts of M.halliana flowers were evaluated by APTT,PT,TT and FIB on plasma of rabbit in vitro.The results indicated that the total extraction,40% methanol fraction,60% methanol fraction,methanol fraction of M.halliana flowers and compound 1-7 exhibited anticoagulant effect in vitro.Subsequently,afzeloside(5),the abundant compound in M.halliana flowers,was investigated antithrombotic effect in vivo.Its antithrombotic mechanisms were evaluated on rats acute blood-stasis model.The antithrombotic effect was evaluated by WBV,PV,HCT,ESR,APTT,PT,TT,FIB,6-keto-PGF1?,TXB2,ET-1 and eNOS in vivo.Afzeloside showed inhibitory effect of thrombus formation,and its antithrombotic effect was related to the regulation of vascular endothelium active substance,activating blood flow and anticoagulant effect.Chapter 2 Dynamic changes in three secondary metabolites of Malus halliana Koehne flowers during bloomingIn this study,the dynamic changes in its secondary metabolites during blooming were investigated.The changes in the contents of three flavonoids(phloretin-4'-O-glycosidase,afzeloside,and 3-hydroxyphloridzin)were determined by high performance liquid chromatography(HPLC).The results indicated that the contents of three flavonoids showed a fluctuating downward trend,thereinto,the maximum contents of phloretin-4'-O-glycosidase,afzeloside,and 3-hydroxyphloridzin reached 157.43±0.36,17.27±0.06 and 22.67±0.35(mg/g),respectively.Chapter 3 Chemical composition and glucose uptake effect on 3T3-L1 adipocytes of Ligustrum lucidum Ait.flowersChemical composition of Ligustrum lucidum Ait.flowers were investigated by using various chromatographic and spectroscopic methods.Seven compounds were isolated and identified from L.lucidum flowers,including ursolic acid(1),kaempferol-7-O-?-L-rhamnoside(2),?-sitosterol(3),?-daucosterol(4),kaempferitrin(5),10-hydroxy oleuropein(6),kaempferol-3-O-?-D-glucopyranoside-7-?-L-rhamnopyranoside(7),compound 1,2,5-7 were isolated for the first time from this flowers,compounds 2,5-7 were isolated for the first time from this plant and compound 2,7 were isolated for the first time in Ligustrum.Then glucose uptake effect on 3T3-L1 adipocytes of L.lucidum extracts and compounds were evaluated.The results showed that 100,33,11?g/mL of total extraction,100,33,4 ?M of kaempferitrin,100,33,11,4 ?M of 10-hydroxy oleuropein and 33,11,4 ?M of kaempferol-7-O-?-L-rhamnoside could increase glucose uptake on 3T3-L1 adipocytes,indicating that L.lucidum flowers possessed potential hypoglycemic activity.