果糖联合酵母诱导大鼠高尿酸血症模型及其尿酸代谢机制研究

发布时间：2018-07-11 15:05

本文选题：高尿酸血症模型 + 酵母　；参考：《青岛大学》2017年硕士论文

【摘要】：目的:随着近年来高尿酸血症患病率的持续升高,高尿酸血症动物模型的建立更加引起了重视,但其造模方法尚未有统一定论,现有的动物模型仍然存在一定的问题。故本研究拟采用酵母和果糖联合的新型造模方法诱导大鼠建立高尿酸血症模型,观察其对大鼠持续性高尿酸血症形成的影响;通过对尿酸代谢途径的研究,初步探讨果糖联合酵母维持大鼠高尿酸水平的机制。方法:选用雄性Wistar大鼠50只,按体质量随机分为五组,每组10只,分别为对照组、酵母组、果糖组、酵母氧嗪酸钾组(高尿酸血症传统模型组)、酵母果糖组。对照组与果糖组给予普通大鼠颗粒饲料喂养,其他三组给予质量分数0.2酵母饲料喂养;果糖组与酵母果糖组大鼠给予10%果糖饮水,其他三组给予自来水;酵母氧嗪酸钾组予以500mg/(kg·d)氧嗪酸钾灌胃。分别在第2、4、6、8周末剪尾取血,测定各组血清尿酸(SUA)、血清尿素氮(BUN)、血清肌酐(SCr)水平。在第8周末时,收集24h尿液,测定尿中尿酸(UUA)、肌酐(UCr),计算尿酸清除率(CUA)和肌酐清除率(CCR)。干预8周后,处死大鼠,检测血清及肝匀浆中腺苷脱氨酶(ADA)和黄嘌呤氧化酶(XOD)活性,并观察肾脏组织学病理切片。采用Western blotting检测大鼠肾脏有机阴离子转运子1(OAT1)、肾脏尿酸盐转运体(RST)、葡萄糖转运蛋白9(GLUT9)的蛋白表达水平。结果:实验第2、4、6、8周末,酵母果糖组大鼠血清SUA水平分别达到345.9μmol/L,403.7μmol/L,447.2μmol/L和451.4μmol/L;与对照组相比分别升高了66.5%、33.8%、37.5%和69.8%(P值均0.05);而酵母氧嗪酸钾组(传统模型组)大鼠SUA水平与对照组相比也显著升高(P值均0.05),分别在实验第2、4、6、8周末升高了28.8%、54.4%、39.4%和75.1%,显示造模成功。在第2周末,酵母果糖组大鼠SUA水平与酵母组、果糖组、酵母氧嗪酸钾组相比显著升高(P值均0.05),分别升高了31.7%、52.9%、29.3%,显示酵母果糖组SUA升高最快;在第8周末,酵母果糖组与酵母氧嗪酸钾组大鼠SUA水平分别是对照组的2.24倍、2.17倍,显示果糖具有氧嗪酸钾类似的促进血清尿酸升高的作用。与嘌呤代谢相关的XOD和ADA活性分析显示,酵母果糖组大鼠血清和肝匀浆中XOD活性分别达到46.3U/L和10.4U/L,明显高于空白对照组(38.2U/L和7.7U/L,P值均0.05);而ADA活力也分别达到26.8U/L和15.3U/L,显著高于对照组(19.5U/L和11.1U/L,P值均0.05)。与对照组相比,酵母果糖组大鼠24h尿量明显增加,CUA明显降低(P值均0.05)。肾脏病理组织学分析显示,在第8周,酵母果糖组大鼠肾小管管腔间质偶见结晶物沉积,无明显纤维化,肾小管、肾小球组织结构无明显异常;酵母氧嗪酸钾组大鼠肾脏偶见间质片灶状单核淋巴细胞浸润,无明显纤维化改变。Western blotting结果显示,与对照组相比,酵母果糖组大鼠肾脏OAT1水平明显下降,RST水平明显升高(P值均0.05)。结论:果糖联合酵母可快速诱发高尿酸血症的出现,引起肾脏炎症反应较小,稳定性与持续性较好,可作为开展嘌呤代谢研究的良好模型;同时,提示摄入高嘌呤食物时应避免果糖的过量摄入。
[Abstract]:Objective: with the increasing prevalence of hyperuricemia in recent years, the establishment of hyperuricemia animal model has become more and more important, but its modeling method has not yet been unified, the existing animal models still have some problems. Therefore, a new model method of combination of yeast and fructose is used to induce the establishment of high uric acid in rats. The effects of hyperuricemia on the formation of persistent hyperuricemia in rats were observed. Through the study of the metabolic pathway of uric acid, the mechanism of sustaining the high uric acid level of rats with fructose and yeast was preliminarily studied. Methods: 50 male Wistar rats were randomly divided into five groups according to the body mass, 10 in each group, which were the control group, the yeast group, the fructose group, and the fermentation. The control group and fructose group were fed with normal rat pellet feed and the other three groups were fed with 0.2 yeast feed, the fructose group and the yeast fructose group were given 10% fructose drinking water, the other three groups were given tap water, and the yeast oxazine potassium group was given 500mg/ (kg. D). The serum uric acid (SUA), serum urea nitrogen (BUN) and serum creatinine (SCr) levels were measured at the end of the 2,4,6,8 week, and the urine acid (UUA) and creatinine (UCr) in the urine were collected at the eighth weekend, and the urine acid removal rate (CUA) and creatinine clearance rate (CCR) were calculated. After 8 weeks, the rats were killed and the serum and liver were detected. The activity of adenosine deaminase (ADA) and xanthine oxidase (XOD) in the homogenate and the histological pathological sections of the kidney were observed. Western blotting was used to detect the organic anion transporter 1 (OAT1), renal urate transporter (RST) and glucose transporter 9 (GLUT9) protein expression level in rats. Results: experimental 2,4,6,8 weekend, yeast fructose group The serum SUA levels of rats were 345.9, 403.7, 447.2, and 451.4 mol/L, respectively, and increased by 66.5%, 33.8%, 37.5% and 69.8%, respectively, compared with the control group (P value was 0.05), while the SUA level of the yeast oxazine group (the traditional model group) was significantly higher than that of the control group (P value was 0.05), respectively, and increased at the end of the experiment at the end of the experiment. 28.8%, 54.4%, 39.4% and 75.1% were successful. At the end of the second week, the SUA level in the yeast fructose group was significantly higher than that in the yeast group, the fructose group and the yeast oxazine group (P value 0.05), which increased by 31.7%, 52.9%, 29.3% respectively, indicating that the yeast fructose group SUA increased the fastest; in eighth weekend, the yeast fructose group and the yeast oxazine potassium group rat SU The level of A was 2.24 times and 2.17 times that of the control group, which showed that fructose had a similar effect on the increase of serum uric acid. The activity of XOD and ADA related to purine metabolism showed that the activity of XOD in the serum and liver homogenate of the yeast fructose group was 46.3U/L and 10.4U/L respectively, which was significantly higher than that of the blank control group (38.2U/L and 7.7U/L, P value). The activity of ADA also reached 26.8U/L and 15.3U/L, which was significantly higher than that of the control group (19.5U/L and 11.1U/L, P value was 0.05). Compared with the control group, the 24h urine volume increased significantly in the yeast fructose group and the CUA decreased significantly (P value was 0.05). The renal histopathology analysis showed that in the eighth weeks, the renal tubule interstitium in the yeast fructose group was crystallized. There was no obvious fibrosis, renal tubules and glomerular structure without obvious abnormalities. The kidney of the group of yeast oxazine group showed interstitial mononuclear lymphocyte infiltration and no obvious fibrosis changes.Western blotting results showed that the OAT1 level of the kidney of the yeast fructose group decreased significantly and the RST level was significantly higher than that of the control group. (P value is 0.05). Conclusion: fructose combined yeast can quickly induce hyperuricemia, resulting in less renal inflammation and better stability and sustainability. It can be used as a good model for the study of purine metabolism. At the same time, it is suggested that the excessive intake of fructose should be avoided when eating high purine food.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R589.7;R-332

【参考文献】