Purification And Physical-chemical Properties Of Superoxide Dismutase From Horsefoot Blood

Objective: To separate and purify the SOD by using an optimal method from horsefoot flotsam blood that comes from the production of horsefoot reagent and mensurate its biological activity,analyse parts of characters of the SOD.Method: SOD of horsefoot blood was isolated and purified by biochemical methods such as calefaction, filtration, Q sepharose F. F ionexchange chromatography and Sephadex G-50 gel filtration respectively, and researched parts of characters of the SOD. The purified SOD and its relative molecular weight were identified by SDS-polyacrylamid gel electrophoresis.Results: SOD of horsefoot blood could be triumphantly separated.This enzyme's livingness was 3849.26U/mg, purification multiple was 62.18, callback rate was 60.63%, the largest ultraviolet absorption wavelength was 264nm, the stability was all right between pH5~9. It could effectively wipe off motley protein and did not influence enzyme activity by heating preservation 20 minutes under 65℃. Checking rarefied SOD by SDS-PAGE, it indicated a single band of SOD and molecular weight of 34kDa.Conclusion: The method has been proved to be effective for the purification of SOD from horsefoot blood , and it can be used to purify plentiful SOD from horsefoot flotsam blood. Objective: To evaluate the safety of Horsefootblood SOD.Methods: The possibility of determining LD50 was judged through pretest.The LD50 or maximum to drug was measured by intragastric administration of the Horsefootblood SOD with the maximum concentration and the maximum amounts in mice.Results: All mice survived,and no toxic reaction was shown in the observed period.The maximum to drug was 1159.2mg of Horsefootblood SOD given to mice per kilogram.Conclusion: No obvious toxicity reactions were observed and Horsefootblood SOD is pretty safe. Objective: To study the anti-aging effect of Superoxide Dismutase from Horsefootblood on D-galactose ( D-ga1) induced aging model mice and the possible mechanism.Method:Ninety-six kunming mice were randomly divided into normal control group,D-ga1 induced aging Model group, vitaminE(VitE) group and superoxide dismutase(SOD)groups.D-gal aging mouse model was established by cervicodorsal region subcutaneous injection with D-gal 120mg/kg once a day for 42 days. In the meantime,drugs were given by intragastric administration respectively in SOD 231.84,115.92,57.96 mg/kg and VitE 100mg/kg treatment groups once a day. The effect of SOD on behavior and form of mice was observed, after 42 days, the SOD, Glutathioneperoxidase (GSH-Px)activity, malondialdehyde(MDA)content in serum,liver,and brain tissues of mice were measured.Results:Compared with normal control group,for aging model mice,the SOD and GSH-Px activities in serum,liver and brain tissues dropped,and the MDA content was raised.Compared with model group,for the mice in SOD and VitE treatment groups,the SOD activity in serum and liver tissue enhanced,the GSH-Px activity i n serum and brain tissue enhanced;for the mice in SOD treatment group, the MDA conten in serum and brain tissue reduced ;for the mice in VitE treatment group,the MDA contenin serum,liver and brain tissues dropped.Conclusion:SOD has anti-aging effect on D-gal induced aging mice, and the action mechanism is related to the increasement of SOD and GSH-Px acfivities,the decreasement of MDA content in serum,liver and brain of D-gal aging mice.