Studies On The Chemical Constituents And Biological Activities Of Bidens Parviflora Willd

Bidens parviflora Willd.,also known as Glossogyne tenuifolia,is a plant of Bidens in Compositae,which is widely distributed in various regions of China.The Dai medicine,called Bidens parviflora as"Yajinbu"and takes the whole herb as medicine,which has a good therapeutic effect on many diseases such as cold,fever,sore throat,malaria,acute gastroenteritis,icteric hepatitis,snake bite and so on.Modern pharmaceutical research shows that the plants of Bidens,such as Bidens pilosa,Bidens cernua,Bidens tripartita,Bidens bipinnata and Bidens biternata,contain flavonoids,phenylpropanoids,caffeinylquinic acids,polyacetylene glycosides and other compounds,which have certain pharmacological activities in terms of lowering blood pressure,regulating blood lipids,protecting cardiovascular and cerebrovascular system,reducing liver damage,anti-inflammatory,antibacterial,antiviral and so on;but there are few studies on the material basis and pharmacodynamics of this plant,at the same time,the traditional biological activity experiments are based on rat or mouse models,making it difficult to investigate the activity of trace monomer compounds in vivo..Therefore,this research will conduct a systematic study on Bidens parviflora.and use the transgenic zebrafish model to screen the corresponding biological activities.Objective to study the chemical constituents of Bidens parviflora Willd.,and to screen the anti-inflammatory activities of the isolated compounds by using the inflammatory model induced by Cu SO4 in zebrafish;to screen the angiogenic activities of the isolated compounds by using the ISV of zebrafish damage model induced by PTK787.Methods the ethyl acetate and n-butanol parts of 80%ethanol extract of Bidens parviflora Willd.were separated and purified by silica gel column chromatography,polyamide column chromatography,ODS column chromatography and PRE-HPLC.The structures were confirmed by ESI-MS?HR-TOF-MS?IR?UV?¹H-NMR?¹³C-NMR?¹H-¹H COSY?HSQC?HMBC?CD and othe identification methods.The directional migration of macrophages in the trunk of zebrafish model induced by Gu SO₄was observed and recorded by the stereofluorescence microscope to investigate the anti-inflammatory activity of the compounds.Use PTK787 to establish a zebrafish intersegmental vessel(ISV)injury model through inhibit the vascular endothelial growth factor(VEGF)and destroy angiogenesis in zebrafish embryos,and then add different concentrations of monomer compounds to intervene,The length of intersegmental vessel was measured by Image Pro Plus 5.0 software,and calculate the ISV growth rate to investigate the angiogenic effect of each compound on the vessel.Results In this experiment,54 compounds were isolated from the 80%ethanol extr act of from Bidens parviflora Willd.According to the physicochemical properties and spectral data of the compounds,51 of them were identified as:(Z)-6-O-(4?-acetyl-6?-O-p-coumaroyl-?-D-glucopyranosyl)-6,7,3?,4?-tetrahydroxyaurone(1),okanin-4-meth yl ether-8?-O-?-D-glucopyranoside(2),okanin-7?-O-?-D-(6?-acetyl)-glucopyranoside(3),Pyridine-4-formyl-O-?-D-glucopyranoside(4),(10R)-tridecane-2E,12-diene-4,6,8-triyne-1-ol-10-O-?-D-glucopyranoside(5),(10R)-tridecane-2E-ene-4,6,8-triyne-1-ol-10-O-?-D-glucopyranoside(6),(10R)-tridecane-2E,8E-diene-4,6-diyne-1,13-diol-12-O-?-D-glucopyranoside(7),4-O-acetyl-3-O-caffeoyl-2-C-methyl-D-erythronate(8),protocatechuic aldehyde(9),6,7-dihydroxycoumarin(10),Protocatechuic acid methy l ester(11),(7'S,8'S)-Sapiumin C(12),trichocarpine(13),syringicacid-4-O-?-L-rham nop yranoside(14),3-O-caffeoyl-2-C-methyl-D-erythrono-1,4-lactone(15),methyl2-O-caffeoyl-2-C-methyl-D-erythronate(16),neochlorogenic acid methyl ester(17),chlorogenic acid methyl ester(18),methyl 4-O-caffeoylquinate(19),5-O-feruloylq uinic acid(20),adenine(21),inosine(22),6,7,3?,4?-tetrahydroxy aurone(23),mariti mein(24),Z-6-O-(6?-diacetyl-?-D-)-6,7,3?,4?-tetrahydroxy aurone(25),Z-6-O-(4?,6?-diacetyl-?-D-)-6,7,3?,4?-tetrahydroxy aurone(26),(Z)-6-O-(4?-acetyl-6?-O-p-coumar oyl-?-D-glucopyranosyl)-6,7,3?,4?-tetrahydroxy aurone(27),okanin-4?-O-?-D-glucop yranoside(28),okanin-4-methyl ether-3?-O-?-D-glucopyranoside(29),okanin-4?-O-?-D-(6?-O-acetyl)-glucopyranoside(30),okanin-4-methylether-4?-O-?-D-(6?-acetyl)-glucopyranoside(31),okanin-4?-O-?-D-(4?-acetyl-6?-trans-p-coumaroyl)–glucopyran oside(32),isookanin(33),isookanin-7?-O-?-D-glucopyranoside(34),homoeriodictyo l-7-O-?-D-glucopyranoside(35),isookanin-3?-methyl ether-7-O-?-D-glucopyranoside(36),isookanin-4?-methyl ether-7-O-?-D-(6?-acetyl)-glucopyranoside(37),7,3?,4?-tri hydroxyflavone(38),luteolin-7-O-?-D-glucopyranoside(39),8,3?,4?-trihydroxyflavor ne-7-O-?-D-glucopyranoside(40),quercitrin-3,3?-dimethoxy-7-O-?-D-glucopyranosi de(41),5,3?-dihydroxy-3,6,4?-trimethoxyflavone-7-O-?-D-glucopyranoside(42),quer cetinn-3,3?-dimethyl ether-7-O-rutinoside(43),quercetin-3,4?-dimethylether-7-O-rutin oside(44),isoquercitrin(45),quercetin-3-O-?-L-arabinoside(46),hyperoside(47),Te tradecane-2E,8E-diene-4,6-diyne-1,14-diol-12-O-?-D-glucopyranoside(48),cordifoli oidyne B(49),bipinnata polyacetyloside B(50),?-sitosterol(51).21 compounds isolated from Bidens parviflora Willd.were screened for their anti-inflammatory activity.The results showed that compounds 18 and 20 had better anti-inflammatory activity,and could significantly reduce the migration of macrophages in the trunk of transgenic zebrafish juveniles.19 compounds were screened for pro-angiogenic activity through the zebrafish ISV injury model,the results showed that compounds 5,7,28,30,48,and 50 had strong activity and could effectively promote the generation of ISV in zebrafish embryos.Conclusion 54 compounds were isolated from Bidens parviflora Willd.and 51 of them were identified.Compounds 1-7 were new compounds,compounds 8 were new natural products,compounds 12-14,20-22,37,48 were isolated from thise genus for the first time.Compounds 25-27,29-32,35,40-44,49-50 were isolated from this plant for the first time.Among the 7 new compounds,compound 1 is a nerone glycoside,compounds 2?3 are flavanone glycosides,compound 3 is an isonicotinic acid glucoside,and compounds 5-7 are polyacetylglycosides.The zebrafish inflammatory model was used to screen the isolated 21 compounds for anti-inflammatory activity.The results showed that caffeoylquinic acid compounds 18 and 20 had better activity,and the activity of compounds 18 and 20was significantly higher than that of other compoundrs at different concentrations.The screening results of 19 monomer compounds by zebrafish intersegmental vessel(ISV)injury model showed that chalcone glycosides 5,7 and polyacetylene glycosides 28,30,48,50 can significantly promote angiogenesis,and with the increase of drugs concentration,the length of intersegmental vessel increases significantly,which can effectively reduce the inhibition effect of PTK787.At the same time.