Study On The Hypoglycemic Material From The Leaves And Twigs Of Taxus Chinensis Var.Mairei

The main purpose of this paper is to screen the hypoglycemic activity fraction from the leaves and twigs of Taxus chinensis var.mairei(Lemee et LevL)Cheng et L.K.Fu.Using HepG2 cells and type 2 diabetic rats as the models,Antidiabetic effects of various extracts from the leaves and twigs of Taxus chiensis var.mairei was investigated in vivo and in vitro.Meanwhile,the n-butanol extract and crude polysaccharides from the leaves and twigs of Taxus chinensis var.mairei was preliminarily investigated.This study could screen preliminarily the hypoglycemic activity fraction and will be helpful for elucidating the fundamental substance of the hypoglycemic activity and the further development.The main research contents and results are as follows:1.The study of chemical constituents from the n-butanol extract of the leaves and twigs of Taxus chinensis var.maireiFifteen compounds were isolated from the n-butanol extract ofthe leaves and twigs of Taxus chinensis var.mairei by using the modern chromatography techniques.And on the basis of the spectroscopic analysis and chemical methods,structures of eight compounds were determined asfollows:Sciadopitysin(1),Berberine(2),(-)-Epicatechin(3),Rutin(4),Myo-inositol(5),E-cinnamic acid(6),Inosine(7),Quercetin-3-O-?-L-arabinopyranosyl(1"'?6")-?-D-glucopyranoside(8),Kaempferol--3-O-?-L-arabinopyranosyl(1'"?6")-?-D-glucopyranoside(9),Kaempferol-3-O-?-L-rhamnopyranosyl-(1'"?6")-?-D-glucopyranoside(10),?-sitosterol-3-O-?-D-glucopyranoside(11).(2),(6),(7),(8),(9),(10)were isolated fromthe leaves and twigs of Taxus chinensis var.mairei for the first time.2.The preliminary study of crude polysaccharides from the leaves and twigs of Taxus chinensis var.maireiCrude polysaccharides were obtained by hot water extraction and ethanol precipitation from the leaves and twigs of Taxus chinensis var.mairei.Phenol-sulfuric acid method is used to determine the sugar content of the crude polysaccharides 30.51%and the refined polysaccharides 62.33%.The Bradford method is used to determine the protein content of the crude polysaccharides 4.52%and the refined polysaccharides 1.78%.Molecular weight determination of polysaccharide of Taxus chinensis var.mairei by gel permeation chromatography column and HPLC-ELSD,with 4 mol·L-1 three trifluoroacetic acid will polysaccharide hydrolysis into simple sugars,determination of the monosaccharide composition of the amino column and HPLC-ELSD.The relative molecular weight of Taxus chinensis var.mairei polysaccharide was in the range of 27.5~115 kDa,composed of galacturonic acid,rhamnose and,arabinose and glucosexylose.The ratio of 5 kinds of monosaccharide was about 2.18:1 3.98:6.85:8.64.3.Component Analysis of leaves oil from Taxus Chinensis var.maireiUsing the yield of leaves oil from Taxus Chinensis var.mairei as the index,investigated the effect of the extraction pressure,extraction temperature,extraction time on the extracting-rate of leaves oil.The optimal parameters of the supercritical CO2 extraction of the oil extracted from leaves of Taxus Chinensis var.mairei were determined.The extracted leaves oil was derivatized with boron trifluoride-methanol complex.33 kinds of fatty acids were identified by GC-MS in oil extracted.The kinds of fatty acids in leaves oil from Taxus Chinensis var.mairei from 3 habitats is compared.4.The effect of different concentrations of each fraction from Taxus chinensis vsr.mairei to the glucose consumption of normol HepG2 cellsThis thesis is to investigate the hypoglycemic activity of different fractions from the leaves and twigs of Taxus chinensis var.mairei using HepG2 cells and insulin resistant HepG2 cells in vitro.The ethanol extract,crude polysaccharides,ethyl acetate extract,and n-butanol extract can increase HepG2 cells and insulin resistant HepG2 cells glucose up take in different extent.0.05 mg·mL-1 and 0.01 mg·mL-1 of samples have the best hypoglycemic effect.5.The effect of different fractions from Taxus chinensis var.mairei on type 2 diabeticThe type 2 diabetic rats are adopted to validate the hypoglycemic activity of the leaves and twigs of Taxus chinensis var.mairei.The ethanol extract and crude polysaccharides show the ability of resisting the diabetes mellitus induced weight-loss,lowering FBC,adjusting the confused blood fats,protecting the viscera and improving glucose tolerance in diabetic rats.