Chemical Constituents Of Physalis Calyx Seu Fructus And Their Biological Activities

Physalis Calyx seu Fructus(PCF)is a traditional Chinese medicine commonly used in the treatment of respiratory diseases in China.It contains diverse chemical components including steroids,flavonoids,terpenoids,alkaloids,phenylpropanoids,sugars,phenolic acids and so on.At present,the chemical compositions and mechanism of PCF in the treatment of respiratory tract inflammation are still unclear.In order to reveal the material basis of the treatment of respiratory diseases,this paper systematically studied the chemical components of PCF.A total of 36 compounds were isolated from the petroleum ether soluble part of PCF by C18 reversed-phase silica gel,silica gel,Sephadex LH-20,MCI gel as well as semi-preparative HPLC.The structures of these compounds determined by means of NMR,MS,UV data analysis and comparison with literatures data.The anti-inflammatory and antioxidant activities of the isolated compounds were evaluated by Nitric oxide(NO)production assay,Dual luciferase reporter gene assay,NAD(P)H:quinone reductase(QR)assay,and Glutathione(GSH)assay.A total of 36 compounds were isolated and purified from the petroleum ether soluble part of PCF,including 2 new compounds and 34 known compounds.Two new compounds is steroids,identified as 25-hydroxy-ergosta-4,24(28)-die-3-one(1)and(24?)-24-hydroxy-ergosta-4,25-die-3-one(2).Thirty-four known compounds including fifteen steroids,physalin L(3),isophysalin B(4),5?-ethoxy-6?-hydroxy-5,6-dihydrophysalin B(5),4,7-didehydrophysalin B(6),4,7-didehydroneophysalin B(7),ergosta-5,25-diene-3?,24?-diol(8),(22E)-5?,8?-epidioxyergosta-6,22-dien-3?-ol(9),gramisterol(10),(3?)-3-hydroxy-26,27-dinorcholest-5-en-24-one(11),26,27-dinorcholest-4-ene-3,24-dione(12),(3?,22E)-3-hydroxy-26,27-dinorcholesta-5,22-dien-24-one(13),physalindicanol B(14),?-sitosterol(15),gitogenin(16),3?-hydroxy-(22E,24R)-ergosta-5,8,22-trien-7-one(17);four alkaloids,aurantiamide(18),isoechinulin A(19),N-benzoyl-L-phenylalaninol(20),aurantiamide acetate(21);four terpenoids,4a?-decahydro-8a?-methyl-4-methylene-6?-(1-methylethenyl)-1?,3?-naphthalenediol(22),capsidiol(23),(+)-anhydro-?-rotunol(24),pubinernoid A(25);five flavonoids,rhamnazin(26),luteolin(27),chrysoriol(28),apigenin(29),eriodictyol(30);six aliphatic compounds,dibutyl phthalate(31),bis(2-ethylhexyl)phthalate(32),1-O-(9Z,12Z-octadecadienoyl)glycerol(33),methyl(10E,12Z)-9-hydroxy-octadecadienoate(34),n-hexadecane acid(35),undecanoic acid(36).Compounds 31 and 32 may be components from organic solvent.Oxidative stress and inflammatory response are important pathogenic factors of respiratory diseases.We evaluated the anti-inflammatory and antioxidant effects of isolated compounds.The results showed that compounds 5-6,19,22-23 and 27 show strong inhibitory activity of NO production;compounds 2-3,8-9,13,24 and 33 show moderate inhibitory activity of NO production.Compounds 6 and 22 relieve LPS-induced cellular inflammation by inhibiting transcription of NF-?B.Compounds 4-7,9,22 and 26 show strong QR induced activity;compound 3 show moderate QR induced activity.Compounds 6,9,22 and 26 significantly induced the transcription of Nrf2.Compounds 6,9,22 and 26 can significantly enhance the level of reduced glutathione in cells in a dose-dependent mannerTwo new steroids and thirty-four known ingredients have been purified from PCF.Importantly,compounds 5-6,19,22-23 and 27 demonstrated potential inhibitory effect against inflammatory response and compounds 4-7,9,22 and 26 were found to be the potential anti-oxidant ingredients of this medicinal plant.These enriched the chemical and biological diversity of PCF.The ethnomedical use of PCF for the treatment of respiratory diseases might be attributed to the inhibitory effects against oxidative stress and inflammatory response by active compound.