| Benign prostate hyperplasia(BPH) is an universal disease among old men.With the trends in aging,the incidence of BPH is stably increasing in China.How to prevent and treat this disease has been an important topic in the medical field for old men.To find safer and more effective agents from natural products attracts more and more attention.In our previous study,we tested over 100 tradition Chinese medicines' extracts by evaluating their inhibition effect on secreted prostate specific antigen(PSA) in LNCaP cells with ELISA method,and found the EtOH extract of Bidens parviflora Willd.showed strong activity.In this thesis,the EtOH extract of Bidens parviflora Wind.was applied to a macroporous adsorptive resins column,and found that the 60%and 30%EtOH eluate are the active fractions of the whole plant,further study on the active constitutes of the two fractions were carried out by bioassay-guided isolation.34 compounds were isolated from the 60%and 30%EtOH eluate of Bidens parviflora Willd by repeatedly applied to silica gel columns,Sephadex LH 20 columns, ODS columns and preparative HPLC.33 compounds' structures were elucidated on the basis of physicochemical property and spectroscopic analysis.The 33 compounds were Benzylβ-D-gluco-pyranoside (1),4-(2-hydroxy-2,6,6-trimethylcyclohexane 4-O-β-D-glucosyl)-3-butylene-2-one(2), 3-(3,4-dihydroxy) phenylpropanoic acid(3),Chromone 7-O-β-D-glucopyranoside(4),(1S,2S)-1-(3-Methoxy-4-hydroxy) phenylglycerol(5),(3R) 8(E)-Decene-4,6-diyne-1,3,10-triol(6*), 5-Hydroxychromone 7-O-β-D-glucopyranoside(7),8(E)-Decene-4,6-diyne-3,10-diol 1-O-β-D-glucopyranoside (8),3,5-Dimethoxyphenylpropylenol 4-O-β-D-glucopyranoside(9),Gallic acid (10),Protocatechuic acid(11),6-Methoxycoumarin 7-O-β-D-glucopyranoside(12),Quercetin(13), Luteolin(14),Kaempferol(15),2'-Hydroxy- 3,4,4',6'-tetramethoxychalcone(16),3,4,2',4', 6'-Pentamethoxydihydrochalcone(17),3,4,2',4'-Tetrahydroxychalcone(18),Acacetin 7-O-(α-D-apiofuranosyl) (1→6)-β-D-glucopyranoside(19),Sulfuretin(20),Maritimein(21),Sulfurein(22), Hyperoside(23),Luteolin 7- O-β-D-glucuronide(24),Kaempferol 3-O-β-D-rutinoside(25),Rutin (27),Quceritrin(28),Kaempferol 3-O-(α-D-arabifuranosyl)(1→2)-β-D-glucopyranoside(29),3,4, 2'-Trihydroxychalcone 4'-O-β-D-glucopyranoside(30),5,3',4'-trihydroxyflavanone 7-O-β-D-gluco-pyranoside (31),Luteolin 7-O-β-D-glucopyranoside(32),((2S,3S) Dihydroquercetin(33), Dihydrolueotin(34) respectively.Compounds 6 was a new compound,twenty-third compounds were isolated for the first time from the Bidens parviflora Willd.Among the 34 compounds,26 compounds showed inhibition effect on PSA secretion in LNCaP cells at different levels.These 26 active compounds belong to three structural types,which were flavanoids,phenyl propanoids and phenolic acids.Flavanoids were more effective than phenyl propanoids and phenolic acids in general.At 15μM test concentration,flavanoids' inhibition rate of PSA secretion in LNCaP cells was about 8.7%to 55.4%,while phenyl propanoids and phenolic acids was about 9.0%to 32.5%.Among these flavonoid compounds,flavone and flavonol showed stronger activity than flavonoid glucoside,while chalcone,flavonone and aurone showed an activity weaker than that of flavone but stronger than that of flavonoid glucoside.In this thesis,with bioassay results in vitro and total contents of flavones as evaluate indexes, the optimal extract method and extract procedure of Bidens parviflora Willd.were firstly studied. The results showed that bioassay results in vitro are parallel with total contents of flavones,the optimal extract method with the best bioactivity and the highest total contents of flavones is to extract by EtOH and then apply to macroporous adsorption resin As an extract procedure,the optimal one is plant material of Bidens parviflora Willd.being extracted by 20 times volume of 60%EtOH under reflux for 3 times,each time lasting 0.5 hour.In this thesis,we first studied on the quality control methods for Bidens parviflora Willd..28 batches of plant materials were collected from different place in domestic,and being used for HPLC fingerprint chromatograph research."Tradition Chinese medicine fingerprint chromatograph similarity evaluation system(China Pharmacopeia 2004A)" and "SPSS" software were used to analyze these chromatographs,which lead to a standard HPLC fingerprint chromatograph.We also studied on the method for simultaneous determination of the contents of 4 active constituents in Bidens parviflora Willd.by Rp-HPLC.Both researches provide quality control method for Bidens parviflora Willd..In conclusion,based on the bioassay evaluation,this thesis studied active constitutes of the raw material plant ofBidens parviflora Willd.,and clarified the main active components of B.P.Willd. for treatment of BPH were flavanoids,phenyl propanoids and phenolic acids.We also studied and established the optimal extract method and procedure of Bidens parviflora Willd.on the basis of bioassay evaluation and content determination of the total flavone.This thesis also studied the HPLC fingerprint chromatograph and determination method of Bidens parviflora Willd.which provide a theory support for the quality control of the material plant when being developed and utilized as a anti-benign prostate hyperplasia medicine.
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