Preliminary Study Of Structure Characterization And Hepatoprotective Effect Of The Dicliptera Chinensis Polysaccharides P1D,P2D And P4D

Dicliptera chinensis was dry whole herb of Dicliptera chinensis(L.)Ness.Mainly produced in Fujian, Taiwan, Guangdong, Guangxi and other places.Sweet-bitter flavor,slightly cold in nature.The main effect is slightly cold liver heat, cooling fluid,dehumidification fire pain, typhus fever. Mainly used for the treatment of hematuria,fever, cough, lung heat cough, combined with membrane inflammation, herpes zoster,jaundice, liver heat headache, unknown swollen poison.2004, Guangdong province Food and Drug Administration for the first time the Dicliptera chinensis sincorporated in the Guangdong Province, Chinese herbal medicine standards, the official Dicliptera chinensis included in the local traditional Chinese medicine standards.Previous work showed that the effective part of Dicliptera chinensis polysaccharide(DCP) was the Mw<6000 Da group. Therefore this paper focuses on the isolation, purification, structure characterization and hepatoprotective effect of the Mw<6000 Da group of DCP.The chief contents and results are showed as follows:1.Isolation and Purification of the effective part of DCPThe effective part of DCP was purified by Diethylaminoethyl DEAE-52 cellulose ion-exchange column chromatography, Sephadex G-50 column chromatography.Purity identify and molecular weight of each sub-components was determinate by gel permeation chromatography(GPC).2. For P1 D,P2D and P4 D of DCP preliminary structural analysisBy infrared spectroscopy(IR), gas chromatography(GC) on P1 D,P2D and P4 D of DCP preliminary structure analysis.3. The protective effect of P1 D,P2D and P4 D of DCP on liver cell line L-02 injury induced by carbon tetrachloride(CCl4).Through detecting L-02 hepatocyte viability, the culture supernatant ALT and AST activity, MDA content and SOD in liver cells and other indicators to evaluate the protective effect of P1 D,P2D and P4 D of DCP on liver cell line L-02 injury induced by CCl4.The results are as follows:1.By DEAE-52,G-50 and GPC, finally obtained three homogeneous polysaccharides P1 D, P2 D and P4 D their molecular weights were 12943 Da, 7261 Da,15259Da.2. Infrared spectroscopy and gas chromatography showed that P1 D was a polysaccharide, and containing β-pyranose ring containing rhamnose, D-arabinose,D-mannose,D-glucose,D-galactose,kinds of monosaccharide molar ratio of1:0.82:0.27:0.33:0.99. P2 D was a polysaccharide, containing rhamnose, D-arabinose,D-glucose, D-galactose, four kinds of monosaccharide molar ratio of 1:3.20:0.99:1.32.P4 D was a polysaccharide, containing rhamnose, D-arabinose, D-mannose, D-glucose,D-galactose, five kinds of monosaccharide molar ratio of 1:0.59:0.55:0.44:0.93.3. Experimental Results showed that P1 D,P2D and P4 D of DCP could significantly increase carbon tetrachloride(CCl4) injury in L-02 liver cell viability,reduce ALT, AST levels, reduce MDA content and increase SOD activity, with obvious effect against CCl4 the role of L-02 induced liver injury. P1 D, P2 D and P4 D dose-response relationship existed, that the higher the concentration of the polysaccharide, the better. Wherein P4 D best effect, P1 D action followed, P2 D worst effects.