| At present,arsenic pollution has become one of the most serious global environmental and public health problems.Arsenic can cause many systemic toxicities,such as immunotoxicity,neurotoxicity,developmental toxicity,and liver toxicity.For arsenic toxicity,nutrition intervention is an effective prevention method.Many studies have shown that selenium can antagonize arsenic toxicity,and that has immune regulation activity.There are some advantages of organic selenium,such as easy absorption,high activity and low toxicity.Grifola frondosa is a kind of medicinal and edible fungus,and its fruiting body has good ability of selenium-enriched.Se-containing protein is the main form of Se in Se-enriched Grifola frondosa,accounting for 57.52% of organic selenium.And the research about Se-containing glycoprotein has not been reported presently.In this paper,the extraction process of water-soluble Se-containing glycoprotein was studied.The crude Se-containing glycoprotein was obtained and purified.The purified fractions were preliminarily characterized.The effects of Se-GPr11 and Se-GPr44 on the immunotoxicity of arsenic induced by NaAsO2 were investigated.It is of great significance to human health.The main researches content are given as follows:(1)Study of extraction for water-soluble glycoprotein from Se-enriched Grifola frondosa.The optimum extraction conditions were as follows: the extraction temperature was 42.3 ?,the extraction time was 7.8 h,the ratio of material to liquid was 1:23.9(g/m L).And the yield was 4.36% under these conditions.(2)Purification and preliminary characterization of water-soluble selenium-containing glycoprotein from Se-enriched Grifola frondosa.4 components were obtained by purification of Se-GPr through DEAE-52 and Sephacryl S-400.The selenium contents were 20.25,10.63,94.01 and 855.27 ?g/g,respectively,and more than non-selenium contents.The results of infrared analysis showed that Se-GPr and its purified components showed similar characteristic absorption peaks compared with the corresponding GPr and their purified components.Se-GPr22 and SeGPr44 were in the range of 1220~1020cm-1 for the CH stretching vibration and 537~606 cm-1 C=O out-of-plane bending vibration was significantly weakened,may be related to the substitution of selenium on sulfur.The amino acid composition of Se-GPr was relatively homogeneous,and Se-GPr11 and Se-GPr44 were in good agreement with the ideal protein conditions.The monosaccharide composition showed in follow: Se-GPr11 consists of arabinose,xylose,mannose,glucose and galactose;Se-GPr22 consists of arabinose,mannose,glucose and galactose;Se-GPr33 and Se-GP44 are composed of mannose,glucose and galactose.Each of the purified selenium glycoproteins is a single component.The molecular weight of Se-GPr11 is 14.32 kDa,Se-GPr22 are composed of two subunits with 20.57 and 31.12 kDa.Se-GPr33 is composed of 15.08,20.57 and 32.78 kDa,and Se-GPr44 consists of three subunits with 16.73,32.78 and 42.46 kDa.(3)Intervention effect and molecular mechanism of water-soluble Se-containing glycoprotein on arsenic-induced immune toxicity.The study found,Se-GPr11 and Se-GPr44 had some protective effects on arsenic-induced immunotoxicity,and prior intervention for 24 h was the best way.Se-GPr11 and Se-GPr44 can antagonize the toxic effects of arsenic by promoting the activity of SOD and inhibiting the production of MDA and ROS.At the same time,Se-GPr11 and Se-GPr44 could down-regulate the expression of p-ERK,p-JNK and p-P38 and contribute to the secretion of IL-2 and INF-? by acting on MAPK channels,to proctect Macrophage RAW264.7 from the immunotoxicity of arsenic induced by NaAsO2. |
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