| Objective: 1.Extraction and spectral analysis of dictyophora polysaccharide:Qualitative and quantitative analysis of dictyophora sporocarp and volva of types and monosaccharide, the two kinds of polysaccharides of serum;2.Effect of dictyophora polysaccharide on Na As O2 by L-02 liver cells intervention:To study the effects of different concentrations of Na As O2 and dictyophora from gene level spectrum effect of polysaccharide on the expression of L-02 in liver cells of all genes by gene chip technology;3.Dictyophora polysaccharide induced liver injury intervention on arsenic:To observe the expression of IKBKG,DUSP6 in liver tissue of rats with arsenic poisoning and treated by dictyophora polysaccharide.And to provide the experimental basis for Dictyophora polysaccharide bioavailability and study the mechanism of arsenic induced liver injury intervention.Methods: 1.Using water extraction and alcohol precipitation of polysaccharides were extracted from sporocarp and volva of sulfuric PMP after derivatization by high performance liquid chromatography analysis of composition and content of monosaccharide;Respectively 100mg/kg dictyophora sporocarp polysaccharide, 100mg/kg dictyophora volva polysaccharide and equal volume of distilled water to rats by intragastric administration, serum was collected after 1h gastric lavage, the determination of the two kinds of polysaccharides in the serum samples of rats in vivo after three chloroacetic acid precipitation;2.Application of Affymetrix Prime View fully human gene expression microarray screening, different concentrations of Na As O2(50, 100, 150 μmol/L) group and polysaccharide(1.6, 8, 40mg/L) intervention differences between Na As O2 exposure L-02 liver cells after gene expression(fold difference is greater than or equal to 5).3.RT-PCR and immunohistochemical analysis were detected in normal group, Na As O2 of different concentration(25, 50, 100mg/kg) group and polysaccharide(25, 50, 100mg/kgl) the relative protein expression and intervention of IKBKG and DUSP6 induced by Na As O2 in rats after liver gene expression of m RNA.Results:1.The detection results of high performance liquid chromatography:dictyphora polysacchride sporocarp and volva respectively containing monosaccharideare:D-mannose,D-glucose,D-galactose,D-xylose,L-fucose,themolarratiois:1.53:52.30:2.78:1.00:1.78;D-mannose,ribose, L-rhamnose,galacturonicacid.D-glucose,D-galactose,D-xylose,L-fucose,13.07:1.00:1.28:189.62:62.74:8.55:31.85.dictyophorasporocarp and volva polysaccharide metabolite in rats in arsenic poisoning peak time and monosaccharide standardconsistency;2.Dictyophora polysaccharide in vitro intervention results:Gene chip analysis results:Compared with the negative control group, and arsenic in the group about 281 screened differentially expressed genes, in the known functional genes, expression is up-regulated in 74, and 107 were down regulated in;dictyophora polysaccharide and gene related difference was 234, among which 6 up-regulated genes, 150 genes were down regulated.With the increased doses of arsenic and the new differentially expressed genes has 346, among them the upregulation of 113, down 91, compared with the arsenic group, with the increase of dictyophora polysaccharide dose and the new differentially expressed genes has 364, which by 107, down 118. 3.Dictyophora polysaccharide in vivo. results: the effect of Na As O2 and the intervention effect of dictyophora polysaccharide on the levels of IKBKG, DUSP6 m RNA expression in rat liver:compared with the control group, the IKBKG m RNA relative expression quantity increased with the doses of arsenic decreased, 50mg/kg, 100mg/kg in arsenic groups obviously difference, the difference was statistically significant(P<0.05);Compared with medium arsenic group,after the treament of different concentration of dictyophora polysaccharide,IKBKG m RNA relative expression of dictyophora polysaccharide increased with the dose increased, which had significant differences between the 50mg/kg, 100mg/kg polysaccharide antagonist group(P<0.05);Compared with the medium arsenic group,IKBKG m RNA group of dictyophora alone relative expression was statistically significant differences(P<0.05).Compared with the control group, the DUSP6 m RNA relative expression quantity increased with higher doses of arsenic and has a trend of decrease, the difference has statistical significance of each arsenic group(P<0.05);Compared with medium arsenic group, DUSP6 m RNA relative expression of dictyophora polysaccharide with the increase of dose increased, which had significant differences between the 25mg/kg, 100mg/kg polysaccharide antagonist group(P<0.05);Compared with the medium arsenic group, DUSP6 m RNA group of dictyophora alone relative expression was statistically significant differences(P<0.05).IKBKG, DUSP6 protein was expressed in different concentration of Na As O2 groups,Compared with the normal group, the difference was statistically significant, the degree of expression and concentration, and increased with the Na As O2 concentration increased with increasing trend.After dictyophora polysaccharide antagonizing, compared with the 50mg/kg Na As O2 group,the expression of IKBKG, DUSP6 protein with the increase of dictyophora polysaccharide has a trend of decline.Conclusion:1.Dictyophora volva polysaccharide containing monosaccharide species and content than dictyophora sporocarp polysaccharide,dictyophora polysaccharide through the mouth into the rats divided into monosaccharides, and have a certain of arsenic poisoning in rats.2.Thegene chip results suggest possible and anti-inflammatory intervention mechanism reaction, Dictyophora polysaccharide on liver injury induced by arsenic enhancement of defense responses, regulation of metabolism.3.The possible mechanism of liver injury and IKBKG, DUSP6 expression of m RNA relative expression quantity and protein are up-regulated induced Na As O2 damage, in addition to the role of dictyophora polysaccharide intervention can improve the Na As O2. |
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