| Xanthine Oxidase (EC.1.2.3.2) is a key enzyme, which catalyze the xanthine andhypoxanthine to uric acid in the metabolism of nucleic acid. During these reactions,the superoxide anion radicals and peroxide are formed. The high concentration of uricacid in body would cause hyperuricemia, and the accumulation of peroxidase wouldinduce tumer, senescent and cardiovascular disease. Therefore, the study of XanthineOxidase inhibitors has garnered widespread attention, recently. However, most ofInhibitors of Xanthine Oxidase have limit application with the sever side effects. So,it’s a new way to extract the components of high inhibitory activity against XathineOxidase from natural algaes.In this reasearch, we made a deep reasearch on the Xanthine Oxidase of UivaPertusa and Gracilaria verrucosa. We confirmed the best optism extraction conditionusing the orthogonal test; with kinetic analysis method, we ensured the inhibitionmechnism, inhibition type and inhibition constant. This study confirmed the freeradical scavenging activity and analysed the chemicals in water solulble extractsfrom two seaweeds. Also, we determined the influence of water solulble extractsfrom Gracilaria verrucosa on mice with hyperuricimia caused by oteracil potassium.The concerte contents and results were as follows:This study optimized the extraction conditions of enthanol extractions of twoseaweeds with orthogonal test and single factor. We also determined the IC50of aceticether and water-soluble components from Uiva Pertusa and Gracilariaverrucosa.The optimum extracting conditions of Uiva Pertusa were extracting time3h, extracting temperature55℃, ethanol concentration70%and the ratio of materialto liquid1:16.Equally, the optimum extracting conditions of Gracilaria verrucosa wereextracting time4.5h, extracting temperature65℃, ethanol concentration90%and theratio of material to liquid1:8. The ethyl acetate and water soluble components wereseparated from the ethanol extract with solvent extracted by ethyl acetate. The resultsdemonstrated that both of the water soluble components had a remarkblely inhibited activity on Xanthine Oxidase. And the IC50of water soluble and ethyl acetatecomponents from Uiva Pertusa and Gracilaria verrucosa were0.58,0.32mg/mL and7.74,0.72mg/mL, respectively. The inhibition kenitic analyzed by Lineweaer Burkplots revealed that the water-soluble extracts from Uiva Pertusa was a uncompetitiveinhibition, and the KISwas0.40. Similarly, the water soluble extracts from Gracilariaverrucosa was a competitive inhibiton, and the KIwas0.84.In this study, we also determined the type of associated inhibion of twowater-soluble components. Cotoxicity coefficient of allppurine and water solublecomponents from Uiva Pertusa was16.2; the typpe was the weak interaction.Cotoxicity coefficient of allppurine and water soluble components from Gracilariaverrucosa was14.6, the type was weak antagonism.The experiment determined the scavenging rate on·OH and O2of thewater soluble copmponents from the two seaweeds. The results were as follows, bothof the water soluble components revealed high scavenging activity on·OH, except onO2.Also, we investigated the chemicals from the water soluble components. Theresults showed that both of them conclude the components of carbohydrate, proteinand alkaloid.In this reasearch, we measured the influence of water-soluble components onmice with hyperuricmia from the two seaweeds. The rsults shows that the mice inhigh dose have a notable effect on lowing the serum uric acid and the weight of micewere steady in the process of this experiment.As normal seaweed along the coast, Uiva Pertusa and Gracilaria verrucosa wereabundant on nutrition and had high inhibited capability on XO, which also conductedas a medicine plant with no side effect. As a result, Uiva Pertusa and Gracilariaverrucosa will have a good prospest for development; It may be become a rawprocessed material on anti hyperuricemia. |
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