| Inonotus obliquus(Pers.:Fr.) Pilat is medicinal fungi of polypores,distributed in the northern hemisphere45°～50°region, Resource is rich in heilongjiang and jilin provinces in our country.Some people used it to treat malignant tumor in the folk. Currently triterpenoids are the main chemical components of Inonotus obliquus, in addition to including steroids, polysaccharides, lignin and small molecule aromatic compounds.Have a lot of pharmacological experiment proves that the Inonotus obliquus have anti-tumor, anti-virus, anti-inflammatory, reduce blood fat, reducing blood sugar and protecting liver and so on a wide range of pharmacological effects.At present, the chemical composition of the systemic study of the Inonotus obliquus is not deep enough, and the activity evaluation of the tiny amounts of chemicals is not comprehensive, in addition, the pharmacological effects of monomer compound more confined to in vitro experiment.Therefore, we studied the chemical composition of the Inonotus obliquus.The sclerotia of Inonotus obliquus.was extracted with95%EtOH,and the extracts was se parated by petroleum ether, ethyl acetate and n-butanol successively to4parts. The extracts was f uther isolated by silica gel, ODS, sephadex LH-20and HPLC chromatograph to obtain the compou nds. The14compounds were isolated,their structures were identified by physicochemical properties and spectroscopy(1H-NMR,13C-NMR, DEPT, HSQC and HMBC).Their names were respectively lanosterol(1),inotodiol(2),3β-hydroxy-lanosta-8,24-diene-21-al(3),inonotsuoxide A(4),inonotsuoxide B(5), trametenolic acid(6),lanosta-8,23E-diene-3β,22R,25-triol(7),betulinic acid(8),3β,28-dihydroxylup-20(29)-ene(9),olelanolic acid(10),4-(3,4-dihydroxyphenyl)-(E)-3-buten-2-one(11),(22E,24S)-5a,8a-epidioxy-24-methyl-cholesta-6,9(11),22-trien-3β-ol(12),8-epidioxy-5α,8α-ergosta-6,22E-dien-3β-ol(13),3β-hydroxylano sta-7,9(11),24-trien-21-oic acid(14),In the14compounds have8tetracyclic triterpene,3pentacyclic triterpene,2steroid compounds, a phenolic compounds, among them10,12and14of compounds isolated from this genus for the first time.The in vitro activity of the separation of Compounds1-10,12-14has carried on the preli minary evaluation using HT-29colon adenocarcinoma cells and HeLa human cervical carcinoma cells.The results showed that compound5showed moderate cytotoxic activity for HT-29cell lin es,the IC50value was22.27μM.compound2,4,8,12,13showed weaker activity,the IC50values were50.31,31.99,59.72,30.76,58.23μM.Compounds4,5on Hela cell lines showed moderate cytotoxic acti vity,the IC50value were12.15,14.22μM,while compound1,2,8,12showed weaker activity,the IC50values were62.29,34.11,30.30,35.82μM.The remaining compounds showed no cytotoxic activity.the results show that compound2,4,5,8,12and13are the active ingredient of Inonotus obliquus.In vivo experiments,to investigate the effects of Compound1,2,6from Inonotus obliquus on oxidative damage induced by CCl4in mice,The activities of superoxide dismutase(SOD), catalas e(CAT), glutathione peroxidase(GSH-PX) and the content of malondialdehyde(MDA) and reductive glutathione(GSH) in serum and liver homogenate were determined.Results showed that the three of the test compound has good antioxidant activity and can protect mice against oxidative stress inju ry induced by CCl4.Among them the activity of compound2is the strongest.In this paper, on the basis of chemical composition and activity in vivo and in vitro stud ies, the experiment clarifies the triterpenoids are the main effective substance of Inonotus obliquu. |
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