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目的:探讨纯化重组猪尿酸氧化酶制剂对实验鸡血尿酸浓度的影响及抗原性。方法:将实验鸡随机分为两组,每组10只。实验组注射重组猪尿酸氧化酶制剂,对照组注射生理盐水,共3次。注射时间分别是实验的d 1,d 7和d 35。注射前及注射后7,24,48,72 h采集血标本,用酶比色法测定实验鸡血清尿酸浓度,间接ELISA测定实验鸡抗重组猪尿酸氧化酶抗体。结果:第1次注射后7,24 h,实验组鸡的血尿酸浓度较注射前分别下降63.38%和28.34%,差异极显著(P<0.01)。第2次注射后7 h尿酸浓度下降52.11%,差异极显著(P<0.01)。第3次注射后血尿酸浓度无显著性下降。对照组注射前后血尿酸浓度比较无统计学差异。3次注射后7 h都能测到尿酸酶活性,酶活性范围1.83～12.75 IU.L-1。第1,2次注射后抗体测定未达到阳性,第3次注射后7 d抗体阳性数为6/10。结论:静脉注射经纯化处理的重组猪尿酸氧化酶能降低实验鸡血清尿酸浓度。该酶可刺激实验鸡产生特异性抗体,该抗体有干扰尿酸氧化酶分解实验鸡血尿酸的作用,但未引起明显过敏反应。
Objective: To investigate the effect of purified recombinant human uroacitase on the concentration of uric acid in experimental chickens and its antigenicity. Methods: The experimental chickens were randomly divided into two groups of 10 rats. The experimental group was injected with recombinant uricase preparations, while the control group was injected with normal saline for 3 times. Injection times were experimentally d 1, d 7, and d 35, respectively. Serum samples were collected before injection and at 7, 24, 48 and 72 h after injection. Serum uric acid concentrations were determined by enzymatic colorimetric assay. Anti-recombinant anti-ubiquitin antibody was tested by indirect ELISA. Results: At 7 and 24 h after the first injection, the serum uric acid concentrations in the experimental group decreased by 63.38% and 28.34%, respectively, before injection (P <0.01). Uric acid concentration decreased 52.11% 7h after the second injection, the difference was significant (P <0.01). Serum uric acid concentration after the third injection no significant decline. The control group before and after injection of serum uric acid concentration was no significant difference. Uricase activity was measured at 7 h after 3 injections, and the enzyme activity ranged from 1.83 to 12.75 IU.L-1. After the first and second injections, the antibody test did not reach a positive level. The positive number of antibodies on the 7th day after injection was 6/10. Conclusion: Intravenous injection of recombinant porcine uroase oxidase can reduce serum uric acid concentration in experimental chicken. The enzyme can stimulate the experimental chicken produce specific antibodies, the antibody interfere with uricase oxidase decomposition of chicken uric acid, but did not cause significant allergic reactions.