Study On The Chemical Components Of Dioscorea Bulbifera And Its In Vitro Anti-lung Cancer Cell A549 And PC-9 Activitie

Objective:"Huangyaozi" is used as medicine with the dried tubers of Dioscorea Bulbifera L..It is bitter,cold and poisonous,and it belongs to the lung and liver meridians.It can be used to resolve phlegm,disperse knots,eliminate galls,clear heat and detoxify.The main components are diterpene lactones,flavonoids and steroids,bibenzyl and phenanthrene quinones.The modern pharmacological research shows that Huangyaozi has anti-tumor,antithyroid disease,antioxidant properties.The purpose of this paper is to separate and purify Dioscorea Bulbifera L.by means of various chromatographic separation methods.and to study the activity of the obtained compounds in human lung adenocarcinoma cells.The optimum extraction process of Dioscorea Bulbifera L.was optimized by taking the content of Diosbulbin B,catechin and Diosgenin as evaluation indexes.Methods:1.Dioscorea Bulbifera L.extract was obtained by reflux extraction with 80%ethanol.The compounds were separated by liquid-liquid extraction,silica gel column chromatography,rapid purification preparative liquid chromatography,semi-preparative high performance liquid chromatography and Sephadex LH-20 gel column,analyzed by MS,13C-NMR and 1H-NMR Identify compound structures.2.CCK-8 method was used to detect the inhibitory effect of the isolated monomer compounds on lung cancer cells A549 and PC-9.3.The effects of solid-liquid ratio,extraction time and ethanol volume fraction on the extraction rate of diosgenin,catechin and xanthin B in Fructus Dioscorea were compared,and the best extraction process of Fructus Dioscorea was optimized.Results:1.Five compounds(compounds 1-5)were isolated from Fructus Dioscorea.They were identified as Diosgenin(1),Diosbulbin B(2),3 αHydroxyl-13 β-Furan-11-one-ape-8-ene-(20,6)-lactone(3),3,5dimethoxyphenol(4),catechin(5).Compound 4 was isolated from Dioscorea Bulbifera L.for the first time.2.CCK-8 detection showed that compared with other compounds,catechin、Diosbulbin B and diosgenin showed strong inhibitory effects on lung cancer cells A549 and PC-9,and their IC50 were 111.3 μmol/L、60.54μmol/L、176.7 μmol/L、142.1 μmol/L and 89.24 μmol/L、170.4 μmol/L.3.Response surface methodology combined with G1 entropy weight method optimized the optimum extraction conditions of Dioscorea Bulbifera L.as follows:extraction time 1.87 h,ethanol volume fraction 47.48%,solid-liquid ratio 1:29.95.Conclusion:Seven compounds were isolated from Dioscorea Bulbifera L.by various chromatographic methods,among which 3,5-dimethoxyphenol was isolated from Dioscorea Bulbifera L.for the first time.In the proliferation inhibition experiments of lung cancer cells A549 and PC-9,Diosbulbin B,catechin and diosgenin showed strong proliferation inhibition effect on lung cancer cells A549 and PC-9.The optimized process method is simple and predictable,the results are reliable,and the evaluation indexes are controllable,which can provide a reference for standardizing the extraction process and quality control of Dioscorea Bulbifera L..