Study On Anti-gout Mechanism Of Peptides Prepared From Marine Fish

Hyperuricemia and gout is one of the common diseases mainly in middle-aged men and post-menopausal women, which is due to long-term high blood uric acid levels, is During the purine metabolism of human body, hypoxanthine and xanthine generated by ATP and nucleic acid through a series of metabolic reactions, and then are hydrolysised to uric acid by xanthine oxidase(XOD). Uric acid is widely spread in human body as a kind of nature antioxidant. Many studies also showed that there are certain associations between uric acid levels and cancer, hypertension, diabetes. Gout drugs available mainly focus on anti-inflammatory and analgesic, such as Benzbromarone can promote excretion of urate. Allopurinol is the only commercially drug which was proven to inhibit the production of uric acid, but it has side effects on the liver and kidneys. So it is very necessary to search for new, safe and without side effects gout drugs.The aim of the present study was to recycle marine fish proteins by enzymatic hydrolysis technologic for functional peptides with anti-gout activity, built a efficient and accurate methods for testing XOD inhibitory activity in vitro, and investigated the effect and mechanism of anti-gout in vivo of marine fish hydrolysates.150μL 0.4mmol/L xanthine and 50μL hydrolysates was hatched at 25℃ for 3 min, and enzymatic the reaction was started with 50μL 0.05U/L XOD and ended with 80μ L 1M HCl. The components was separated by HPLC, the quantitative HPLC separations were performed at a temperature of 25℃ on a Agilent XDB C18 reversed-phase column(250×4.6 mm, 5 μm particle size). The mobile phase was 90% 15 mmol/L ammonium dihydrogen phosphate solution and 10% methanol. The flow rate was 1 mL/min and the absorbance detector was set at 290 nm.Optimum hydrolysis conditions of mulriple enzymes were obtained by single factor experiment and response surface methodology. Marine fish protein was hydrolyzed by 0.16% Neutral protease and 0.13% trypsin for 5.99 h at 55℃ and pH 7.0. The XOD inhibition activity and protein recovery of hydrolysate was 32.25% and 72.75%, respectively.The hyperuricemic animal model was induced by potassium oxonate, and it’s uric acid levels was reduced after the treatment of optimized marine fish hydrolysates for 30 days. The marine fish hydrolysates acted as XOD inhibitors in vivo, and it could drop the expression of mRNA of ADA and XOD, the content of urea nitrogen and creatinine was decreased as the same time.In summary, the marine fish hydrolysates had a good anti-gout activity, the functional polypeptide was a good source of potential drugs for hyperuricemia and gout. There were practical significance and economic value for its study and production.