Bio-Assay Guided Isolation Of The Anti-Depressive Constituents Of Polygala Japonica Houtt.& Studies On The Active Constituents Of Patrinia Scabra

Polygala japonica HOUTT. (Polygalae), widely distributed in China, was used as anti-depressant and memory-and-learning-function-improving agent in folk medicine. This dissertation describes the chemical and pharmacology study of this plant, especially the bio-assay guided isolation of anti-depressive compounds. Some of pure compounds from this plant had been tested for the activity of neurons regeneration, anti-inflammatory and anti-tumor in vitro. The method of the quality control of the material had also been established.The EtOH extract of P. japonica, n-Butol fraction and total saponin showed sound anti-depressive activity on Forced Swimming Test (FST) and Tail Suspension Test on mouse. 15 compounds were obtained from n-Butol extract, the active part, by a variety of chromatography methods, and their structures were established on the basis of spectral data including UV, IR, EI-MS, ESI-MS, 1D-NMR, 2D-NMR, physical and chemical properties, as follows: PJ-29 : 3,5-dihydroxy-7,4'-dimethoxyflavone-3-O-β-D-apiofuranosyl(1→2)-β-D-galactopyranoside (polygalin B); PJ-30 : 3,5,3'-trihydroxy-7,4'-dimethoxyflavone-S-O-β-D-apiofuranosyl(1→2) -β-D-galactopyranoside (polygalin C); PJ-31: polygalasaponin D; PJ-32 : polygalasaponinⅡ; PJ-33: polygalasaponin Ⅲ; PJ-34: polygalasaponin Ⅶ; PJ-35: polygalasaponin E; PJ-36: polygalasaponin F; PJ-37: polygalasaponin G; PJ-38: polygalasaponin H; PJ-39: polygalasaponin XXI; PJ-40: polygalasaponin Ⅴ; PJ-41: polygalasaponin J; PJ-42: polygalasaponin X. Among them, PJ-35 and PJ-38 demonstrated significant anti-depressive activity which reduced the immobility status by more than 50% on forced swimming mouse. It was found that the crude saponins of Polygala japonica and PJ-38 reduced the immobility status of forced-swimming rats in a dose-dependent manner.The chemical investigation of other extracts of this plant resulted in the isolation of 32 compounds and 28 structures of them were determinated as follows also counted on the spectral analysis and physic-chemical properties: PJ-1 : 1,7-dihydroxy-3,4-di- methoxyxanthone; PJ-2: kaempferol-7,4'-dimethyl ether; PJ-3:physcion; PJ-4: castaprenol-12; PJ-5: P-sitosterol; PJ-6: stigmasterol; PJ-7: stigmasta-7,22-di-ene-3-one; PJ-8: n-dotriacontanol; PJ-9: rc-hexadecanol; PJ-10 arachidic acid; PJ-11: erogosterol; PJ-12: l,6,8-trihydroxy-2,3-methylenedioxo-5-geranylxanthone; PJ-13: 5,3'-dihydroxy-7.4'-dimethoxyflavonol; PJ-14: rhamnetin; PJ-15: kaempferol; PJ-16: quercetin; PJ-17: daucosterol; PJ-18: p-hydroxybenzoic acid; PJ-19: coumaric acid; PJ-20: l-0-J3-D-glucopranosyl-(2S,3S,4R,llE)-2-[(2'R)-2'-hydroxypalmitoylamino]-l 1 -octadecene-1,3,4-triol (polygalacerebroside); PJ-21: 3,5-dihydroxy-7,4'-dimethoxyflavone-3-0-/?-D-galactopyranoside (polygalin A); PJ-22: rharnnocitrin3-0-y#-D-galactopyranoside; PJ-23: 3,5,3'-trihydoxy-7,4'-dimethoxy-flavone-3-0-/?-D-galactopyranoside; PJ-24: astragalin; PJ-25: 2p\27-bihydroxy-23-carboxyoleanolic acid 3-O-p-D-glucopyranoside; PJ-26: rhamnetin 3-0-/?-D-galactopyranoside; PJ-27: quercetin 3-0-/?-D-glucopyranoside; PJ-28: rhamnetin 3-O-/?-D-glucopyranosideoAmong all the compounds mentioned above, PJ-12, PJ-20, PJ-21, PJ-29, PJ-30, PJ-35, PJ-36, PJ-37, PJ-38 and PJ-41 were new compounds, andPJ-1, PJ-2, PJ-3, PJ-4, PJ-6, PJ-7, PJ-8, PJ-9, PJ-10, PJ-11, PJ-13, PJ-14, PJ-18, PJ-19, PJ-23, PJ-25, PJ-26, PJ-27 were isolated from this plant for the first time.In analysis of some selected saponins' function, PJ-39 and PJ-40 showed the reverse effects against myelin's neurite outgrowth inhibition power at nanomolar concentrations in vitro, and PJ-38 displayed significant anti-inflammatory activity in vivo with asprin as control.The anti-tumor activity of the total extract of P. japonica, five crude extracts of this plant and some pure compounds were also evaluated by MTT assay, a short term in vitro test. The results indicated that compound PJ-12 showed cytotoxic effects against 5 human cancer cell lines, such as K562^ A549, PC-3M> HCT-8, SHG44.Compounds PJ-35, 38 and 40 showed significant bio-activities in the pharmacology studies, which could be used as marker substance for quality control of P. japonica. So we developed a simple, precise and reproducible HPLC-UV method for the detection and quantification of these active compounds in medicinal material...