Studies On The Chemical Constituents Of Bupleurum Chinese DC.and Ziziphus Jujuba Mill.and The Exploration Of The Active Ingredients Of Xiao Chai Hu Decoction For Application To Depression Based On Network Pharmacology

Xiaochaihutang(XCHT),a famous Chinese herbal formula,has been used clinically in depressive disorders in China.It consists of seven crude drugs:Bupleurum chinese D C.;Scutellaria baicalensis Georgi.;Panax ginseng C.A.Meyer.;Glycyrrhiza uraiensis Fisch.;Pinellia ternata(Thunb.)Breit.;Zingiber officinal Rosc.and Ziziphus jujuba Mill.However,the actual bioactive components of XCHT have not been elucidated.Base on the previous studies,we have investigated the chemical constituents of the 75%ethanol extract of the roots of B.chinense.and Ziziphus jujuba Mill.At the same time,the network pharmacology approach has been used to to determine active gredients and potential targets of XCHT for treatment of depression.Bupleurum chinense D C.,customarily called "Bei-Chai-Hu" in China and included in the Chinese pharmacopoeia,is distributed mainly in Hebei,Liaoning,Jilin and Heilongjiang provinces of China.Its roots have been used as a Chinese herbal medicine for the treatment of fever,influenza,malaria,distending pain in the chest and menstrual disorders for approximately 2000 years.By means of various chromatographic methods such as silica gel.sephadex LH-20,ODS column chromatography and semi-preparative HPLC,70 compounds were isolated from the 75%EtOH extract of Bupleurum chinense D C.Their structures were established by direct interpretation of their spectral data,mainly HR-ESI-MS,1D-NMR and 2D-NMR,CD,as well as by comparison with literature data.Therefore,the compounds were identified as 3?,16?,23,28-tetrahydroxy-11?-methoxy-ole-an-12-ene 3-O-?-D-fucopyranoside(1).3?,16?,28-trihydroxyl-11?-methoy-olean-12-ene 3-O-?-D-fucopyra noside(2),3?,16?-dihydroxy-23-O-acetyl-13?,28-epoxy-olean-11-ene 3-O-?-D-fucopyra noside(3),3?,23,28-trihydroxy-11,13(18)-diene-16-one 3-O-?-D-glucopyranosyl-(1?3)-?-D-fucopyranoside(4),16?,22?,23?,28?-tetrahydroxy-11?-methoxyl-olean-12-en-3-one(5),16?,23?,28?trihydroxyoleana-11,13(18)-diene-3-one(6),3?,23?,28?-trihydroxyoleana-11,13(18)-diene16-one(7)saikolignanoside A(8),saikogenin F(9),saikogenin G(10),saikogenin D(11),saikogenin A(12),prosaikogenin G(13),prosaikogenin F(14),prosaikogenin A(15),prosaikogenin D(16),pleurosaponin F(17),saikosaponin-a(18),saikosaponin-d(19),saikosaponin-e(20),2"-O-acetylsaikosaponin-a(21),3"-O-acetylsaikosaponin-a(22),6"-O-acetylsaikosaponin-a(23),23-O-acetylsaikosaponin-a(24),23-hydroxy-13?,28?epoxy-olean-11-ene-16-one 3-O-?-D-glucopyranosyl-(1?3)-?-D-fucopyranoside(25),6"-Oacetylsaikosaponin-d(26),saikosaponin-c(27),saikosaponin-b1(28),saikosaponin-b2(29).2"-O-acetylsaikosaponin-b2(30).6"-O-acetylsaikosaponin-b2(31),saikosaponin-b3(32),saikosaponin-b4(33),6"-O-acetylsaikosaponin-b4(34),bupleuroside ?(35),saikosaponin-t(36),11?-methoxy-saikosaponin-f(37),saikosaponin-f(38),((?)8R)-9'-methoxyl-donydrodiconiferylalcohol 4-O-?-D-glucopyra noside(39),acutissimalignan B(40).salicifoline(41),pluviatolide(42),guamarolin(43),(+)-pinoresinol(44),(+)-nortrachelogenin(45).2-(3"-methoxy-4"-hydroxybenzyl)-3-(3'-methoxy-4'-hydroxyl-benzyl)-?-butyrolact-one(46),?-spinasterol(47),?-spinasterol-?-D-glucoside(48),oleanolic acid(49),?-Amyrin(50),ursolic acid(51),ursodeoxycholic acid(52),eugenin(53),2,5-dimethyl-7-hydroxycheomone(54),saikochrome-A(55),1-(2,4-dihydroxyphenyl)-3-hydroxy-3-(4-hydroxyphenyl)-1propanone(56),isorhamnetin(57),luteolin(58),quercetin(59),rutin(60),vitexin-2-O-rhamnoside(61),kaempferol-3-?-D-(6-O-trans-p-coumaroyl)glucopyranoside(62),isorhamnetin3-O-rutinoside(63),kaempferol-3-O-?-D-glucopyranoside(64),?-viniferin(65),methylpyroglutamate(66),vanillic acid(67),isoacteoside(68),martynoside(69),apigenin-6,8-diC-?-D-glucopyranoside(70).Among them,compounds 1-8,39 were new compounds and compound 25 was isolated from the natural product for the first time.In general,compounds 3,17-19,21-23,30,42,46 showed more potential cytotoxic activities against all the selected cell lines than the positive control 5-fluorouracil.Based on the primary screening results,the preliminary structure-activity relationship(SAR)studies were also discussed.The SAR results suggest that the 13,28-epoxy bridge,the orientation of the hydroxyl group and the type of the sugar units are important requirements for cytotoxicity and selectivity.Meanwhile,compounds 5-7,11-12 showed significant neuroprotective effects against H2O2-induced SH-SY5Y cell death.The antioxidant and cytotoxic effects of isolated lignans were evaluated in vitro.All compounds exhibited moderate ABTS free radical scavenging properties.Ziziphus jujuba Mill.,a thorny Rhamnaceous plant,has also been used as a traditional Chinese medicine(TCM)for the treatment of anorexia,fatigue,and loose stools in deficiency syndromes of the spleen.By means of various chromatographic methods and semi-preparative HPLC,25 compounds were isolated from the 75%EtOH extract of Ziziphus jujuba Mill..Their structures were established by direct interpretation of their spectral data,mainly HR-ESI-MS,1D-NMR and 2D-NMR,CD,as well as by comparison with literature data.Therefore.the compounds were identified as(7R,8R,7'R,8'S)-guaiacylglycerol-?-O-4'dihydrodisinapyl ether(71),(7S,8R,7'R,8'S)-guaiacylglycerol-?-O-4'-dihydrodisinapyl ether(72),(7S,8S,7'R,8'S)-guaiacylglycerol-?-O-4'-dihydrodisinapyl ether(73),7R,8R-guaiacylglycerol 8-(4-hydroxymethyl-2-methoxyphenyl)ether(74),7R,8S-guaiacylglycerol 8-(4hydroxymethyl-2-methoxyphenyl)ether(75),(7',8'-threo,7R,8S)-dihydroxydehydrodiconiferyl alcohol(76),7R,8S-dehydrodiconiferyl alcohol(77),7S,8R-dehydrodiconiferyl alcohol(78),7',8'-threo,7S,8S-1-[4-[(2-hydroxy-2-(4-hydroxy-3-methoxyphenyl)-1-(hydroxy-methyl)ethoxy]-3-methoxyphenyl]-1,2,3-propanetriol(79),leptolepisol Z(80),erythro-2,3-bis(4-hydroxy-3-methoxy-phenyl)-3-methoxypropanl(81),(1S,3S)-1-methyl-1,2,3,4-tetrahydro-?-carboline-3-carboxylic acid(82),(1R,3S)-1-methyl-1,2,3,4-tetrahydro-?-carboline3-carboxylic acid(83),grasshopper ketone(84),dihydrophaseic acid(85),methyl(3R,5R)-5methoxy-3-phenylisoxazolidine-5-carboxylate(86),3',4',5'-trimethoxycinnamyl alcohol(87),2-{4-[(1E)-3-Hydroxyprop-1-en-1-yl]-2,6-dimethoxyphenoxy}propane-1,3-diol(88),ferulic acid(89),vomifoliol(90),(p-hydroxyphenyl)glycol(91),2"-glucosyl-8-C-glucosyl-4'-Omethylapigenin(92),6"'-feruloylspinosin(93),zizybeoside ?(94),(-)-lyoniresinol(95).Among them,compounds 75,80 are new compounds.In order to further investigation of the antidepression effects of Ziziphus jujuba Mill..The neuroprotective effects of compounds 71-95 against corticosterone-induced neuronal cell damage in human neuroblastoma SH-SY5Y cells were evaluated.Compounds 71,78-80.92 showed significant neuroprotectiv e effects against corticosterone-induced SH-SY5Y cell death.In this work,network pharmacology was employed to identify key compounds and potential targets,and to uncover the molecular mechanism of XCHT on depression.The present work provides an alternative strategy for the deep investigation and understanding of the chemical and pharmacological basis of TCM,which will promote drug discovery from herbal medicines.