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Objective: Preparation of Limulus reagent waste generated plasma as raw material, the best way to extract the superoxide dismutase (SOD), purified to electrophoretic homogeneity horseshoe crab blood SOD, and to analyze and identify some of the physical and chemical properties. Methods: heat denaturation, ultrafiltration, Q Sepharose FF ion exchange column chromatography and purified by Sephadex G-50 gel filtration and other biotechnology separation SOD, and some of its properties analyzed and identified. SDS-polyacrylamide gel electrophoresis to determine its relative molecular weight and its purity. Results: separated successfully from horseshoe crab blood SOD, enzyme specific activity 3849.26U/mg, a purification factor of 62.18, the recovery rate was 60.63%, the maximum UV absorption wavelength of 264nm buffer pH in the range of 5 to 9 good enzyme stability, heated within 65 ℃ 20min, can effectively remove the contaminating proteins without affecting enzyme activity. Horseshoe crab blood SOD purified by SDS-PAGE analysis shows a single band of molecular weight of about 34kDa. Conclusion: This method can be effectively isolated from horseshoe crab blood purified SOD of SOD can be used to extract from horseshoe crab blood of large quantities of waste. Purpose: extract of horseshoe crab blood SOD mice administered Acute Toxicity Test, to make a preliminary evaluation of its safety and to provide a basis for clinical medication safety. Methods: by preliminary experiments to determine the possibility of determination of median lethal dose (LD50), which determined the LD50 or the maximum dose. Results: Pre-experiment ig all animal health deposit, no any toxicity, maximum dose trial. The maximum dose of horseshoe crab blood SOD 1159.2mg/kg. Conclusion: The horseshoe crab blood SOD no apparent toxicity, indicating that the higher the safety of this product. Objective: To study horseshoe crab blood superoxide dismutase antioxidant effect and its mechanism of D-galactose model mice. Methods: Kunming mice 96, were randomly divided into blank control group, aging model group, the vitamin E group, horseshoe crab blood SOD, in the low-dose group, 16 in each group. In addition to the blank control group, each daily injections with D-galactose the 120mg/kg neck back, continuous administration 42d, horseshoe crab blood SOD in the low-dose group were administered daily administration 231.84mg/kg , 115.92mg/kg, 57.96mg/kg, vitamin E group ig 100mg/kg. Observed the mice in each group behavior, morphology changes, and 42d after using the kit to detect serum, liver, brain, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) activity and the content of malondialdehyde (MDA). The result: the detection of biochemical indicators in mice, the aging model group compared with the control group, serum, liver, brain SOD, GSH-Px activity decreased MDA content increased. Compared with the aging model group, the horseshoe crab blood SOD and vitamin E administered mice treated with serum, liver SOD activity and serum, brain GSH-PX vitality significantly increased SOD activity in the brain and liver GSH-PX vitality No significant changes; SOD serum levels of horseshoe crab blood and brain MDA levels were significantly reduced, no significant difference in MDA content in the liver; vitamin E serum, liver, brain MDA levels were significantly decreased. Conclusion: The horseshoe crab blood SOD aging model mice induced by D-galactose good resistance to oxidation, can significantly improve the mice SOD, GSH-PX levels and MDA.
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