Tsugeki Flavonoids And Anti-tumor Activity

Purpose:1. The study is started by extracting and isolating the chemical constituents from the EtoAc part and butanol part, and give reference for studies on the bioactive constituents of Cudrania tricuspidata.2. To optimize the extraction technology for total flavonoids and total polysaccharides from Cudrania tricuspidata, and give reference for development and utilization of Cudrania tricuspidata; To determine cycloartocarpesin in Cudrania tricuspidata from different areas by HPLC, and give new reference for quality evaluation standard.3. Through vitro Anticancer activity screening of total flavonoids and total polysaccharides of Cudrania tricuspidata. enlarge application rang, and give reference for further study of its anticancer mechanism.4. To study the purification process of total flavonoids in Cudrania tricuspidata with macroporous resin, and give science referenc for industrial production.Material and method:1. Material:Cudrania tricuspidata (Carr.)Bur is the dried root and rhizome of Moraceae plants, which is collected from Guangxi.The kind of commercial drugs is collected from Guangxi and Anguo.2. Method:(1) Isolation and Purification monomeric compound from the EtoAc part and butanol part by chromatographic methods such as silica gel, and Sephadex LH-20.(2) Optimize the extraction technology for total flavonoids and total polysaccharides from Cudrania tricuspidata by single factor test and orthogonal test; Determine cycloartocarpesin in Cudrania tricuspidata from different areas by HPLC.(3) MTT assay was employed to investigate the effects done by total flavonoids and total polysaccharides on the growth and proliferation ability of HeLa,A549,HepG2 and SGC-7901 cells.(4) Using static absorption to optimum maeroporous resin which was used for purifying total flavonoids in Cudrania tricuspidata. Using dynamic absorption to the condition of purifying total flavonoids of Cudrania tricuspidata by macroporous resin.Results:1. Seven monomers have been isolated form the EtoAc part and butanol part. They are ①Dihydromorin,②Daucosterine,③5,7,4'—trihydroxyldihydroisoflavone,④Kaempferol,⑤Quercetin,⑥β-sitoste-rol,⑦cycloartocarpin.2. By single factor test and orthogonal test, the optimum process of total flavonoids was as follows:the crude drug was extracted three times, the 60% alcohol density,12 times alcohol was added and 1.5 h every time; The optimal technology conditions of total polysaccharides were:extracting temperature was 90℃, liquid-to-solid ratio was 10:1, extracting time was 3 h. HPLC method was established. Column:Aglinet C18 (150×4.6mm,5μm), mobile phase: Acetonitrile-water as gradient eluent, flow rater:1.0 ml·min-1, temperature:25℃, detective wavelength:203 nm.3. Total flavonoids have more inhibitory effects on viability of HeLa cells,A549 sells,HepG2 cells and SGC-7901 cells than negative control group; but total polysaccharides have less inhibitory effects than total flavonoids.4. Optimum maeroporous resin was AB-8, and the optimum condition was as follow:the speed that sample flows through macroporous resin was 1.5 mL·min-1, and the total flavonoids absorbed on maeroporous resin can eluted by 75% ethanol of 5 BV.Conclusion:1. Six monomers have been isolated form the EtoAc part and butanol part. They are①Dihydromorin,②Daucosterine,③5,7,4'—trihydroxyldihydroisoflavone,④Kaempferol,⑤Quercetin,⑥β-sitoste- rol,⑦cycloartocarpin.2. These extraction technology of total flavonoids and total polysaccharides are reasonable and feasible, which can be used as experimental basis for extraction technology for total flavonoids. The research determined cycloartocarpesin in Cudrania tricuspidata by HPLC firstly, and the method is simple, sensitive, and highly reproductive, and give new reference for quality evaluation standard.3. Total flavonoids have more inhibitory effects on viability of HeLa cells,A549 sells,HepG2 cells and SGC-7901 cells than negative control group, and inhibitory effects depend on drug's concentration. But total total polysaccharides have less inhibitory effects than total flavonoids.4. Optimum maeroporous resin was AB-8, and the optimum condition was as follow:the speed that sample flows through macroporous resin was 1.5 ml·min-1, and the total flavonoids absorbed on maeroporous resin can eluted by 75% ethanol of 5 BV.