Study On The Chemical Constituents And Biological Activities Of The Water Fraction From The Fibrous Roots Of Anemarrhena Asphodeloides Bge.

Objective:By separating the chemical components of the water-soluble parts of the fibrous roots of Anemarrhena asphodeloides Bge.,and some of the compounds were subjected to preliminary activity studies.We hope to provide a theoretical basis for the development and utilization of the fibrous roots of A.asphodeloides Bge..Method:The dried fibrous roots of A.asphodeloides Bge.were pulverized and extracted with 70%ethanol.Separation and purification were carried out by various column chromatographic（CC）separation techniques（silica gel,Sephadex LH-20,Rp-18 reverse phase,High-pressure semi-preparative liquid chromatography,etc.）.The structure of the isolated compounds was identified by modern spectroscopy.To investigate the protective effect of some compounds isolated from the fibrous roots of A.asphodeloides Bge.on SH-SY5Y cells.A preliminary study on the anti-hepatocarcinoma mechanism was tested.Results:Seventeen compounds were isolated from the fibrous roots of A.asphodeloides Bge.,and the chemical structures were identified.The results were followed:4-[Formyl-5-（methoxymethyl）-1H-pyrrol-1-yl]butanoate（1）、perlolyrine（2）、7-Hydroxy-3-（4-methoxybenzyl）-chroman（3）、Methyl 4-hydroxybenzoate（4）、4′-hydroxyphenyl ethyl vanillate（5）、1,3,6,7-tetrahydroxy-9H-xanthen-9-one（6）、Methyl 2-（2,4-dihydroxy-3-（4-hydroxybenzoyl）-6-methoxyphenyl）acetate（7）、6’,7’-dihydroxy-2-methoxy-3’,4’-dihydro-2’H-spiro[cyclohexane-1,1’-naphthalene]-2,5-dien-4-one（8）、Michehedyoside D（9）、Demethyleugenolα-D-glucopyranoside（10）、4′,6-dihydroxy-4-methoxybenzophenone-2-O-（2′′）,3-C-（1′′）-1′′-desoxy-α-L-fructofura noside（11）、syringaresinol-4’-O-β-D-glucoside（12）、（2,6-dihydroxy-3-（（（1S,4S,6R）-4-hydroxy-3,7-dioxabicyclo[4.1.0]heptan-4-yl）methyl）-4-methoxyphenyl）（4-hydroxyp henyl）methanone（13）、（S）-5-（2-hydroxy-5-（3-（4-hydroxyphenyl）propyl）-4-methoxyphenyl）pyrrolidin-2-one（14）、（4-hydroxyphenyl）（2,3,4-trihydroxyphenyl）methanone（15）、（S）-5-（2,4-dihydroxy-3-（4-hydroxybenzoyl）-6-methoxyphenyl）pyrrolidin-2-one（16）and 2′,3′-dihydroxyl-osthole（17）.Effects of neuroprotection of compounds 7,13 and 14 were performed.The results of the MTT assay showed that compounds 7 and 13 showed protective effects on H₂O₂-injured SH-SY5Y cells at 25μM,and the effect also showed a decreasing trend with increasing concentration.We investigated the effects of compounds 7,11,13,14,15 and 16 on cell viability of Hep G2 and Hep 3B cells.All tested compounds showed a significant role in inhibiting the growth of Hep G2 and Hep 3B cells,and they were significantly higher than the 5-FU group.Among them,compound 16 had the most significant inhibitory effect on Hep G2 cells,and its IC₅₀ value was 0.15μM.Therefore,we further investigated the mechanism of compound 16 on apoptosis of Hep G2 cells.Compound 16 could promote apoptosis of Hep G2 cells in a dose-dependent manner.Compound 16up-regulated the expression of Cleaved Caspase-3,Cleaved PARP and Bax in a dose-dependent manner,and down-regulated the expression of Bcl-2.It also could down-regulate the expression of total cellular p65 and nuclear p65.Conclusion:We searched by sci-finder to find that compounds 7,13,14,16 were new compounds,and compounds 1,2,4,6,8,9,10,12 and 17 were found firstly from the fibrous roots of A.asphodeloides Bge..The results of the MTT assay showed that compounds 7 and 13 showed protective effects on H₂O₂-injured SH-SY5Y cells at 25μM,and the effect showed a decreasing trend with increasing concentration.All tested compounds showed inhibition effect on the growth of Hep G2 and Hep 3B cells.Compound 16 may promote the occurrence of apoptosis by limiting cytoplasmic nuclear translocation of p65.