Intestinal Flora in Uricase-Deficient Rats
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摘要:
目的 观察雄性尿酸酶缺失大鼠（Kunming-DY，即KDY大鼠）肠道和新鲜粪便的菌群变化。 方法 与同性别同日龄野生型SD大鼠对照，选取45日龄雄性尿酸酶缺失大鼠称体质量，乌拉坦麻醉后解剖取血制备血清，将小肠依次分成18段，结肠分成2段（每段5 cm）。用磷钨酸法检测血清尿酸含量，用DNA提取试剂盒提取各肠段和新鲜粪便DNA，采用特异性16s rDNA引物扩增后进行高通量测序，将测序结果与数据库比对在门、纲、目、科和属水平鉴定菌群。 结果 45 d日龄时，KDY大鼠体质量下降（P < 0.05），但血尿酸水平明显升高（ P < 0.05），肠道菌群多样性呈降低趋势（ P < 0.05），但粪便菌群多样无明显变化（ P > 0.05）。KDY大鼠肠道和粪便中乳酸杆菌属丰度明显增加（ P < 0.05）。 结论 相比野生型SD大鼠，KDY大鼠肠道菌群多样性下降，但肠道和粪便中的乳酸杆菌属丰度明显增加。 -
关键词:
- 尿酸酶缺失 /
- Kunming-DY大鼠 /
- 高尿酸血症 /
- 肠道菌群 /
- 乳酸杆菌属
Abstract:Objective To investigate the intestinal flora in male uricase-deficient rats （Kunming-DY, KDY rats） compared with wild-type SD rats of the same sex and age. Methods 45-day-old male uricase-deficient rats were weighed, and their blood was collected to obtain serum after they were anesthetized with uratan. Their small intestine was divided into 18 segments, and the colon was divided into 2 segments （5 cm each）. The serum uric acid （SUA） was detected by phosphotungstic acid method. DNA in intestinal segments and fresh feces was extracted using DNA extraction kit. High-throughput sequencing was performed after amplification with specific 16S rDNA primers. Results At 45 days of age, the body mass of KDY rats decreased （P < 0.05）, but the SUA level increased significantly （ P < 0.05）. The intestinal microbiota diversity in KDY rats decreased （ P < 0.05）, but their fecal microbiota diversity did not change significantly （ P > 0.05）. The abundance of Lactobacillus in intestinal tract and feces of KDY rats was significantly increased （P < 0.05）. Conclusion Compared with wild-type SD rats, the intestinal microbiota diversity of KDY rats decreased, but the abundance of Lactobacillus in intestinal tract and feces increased significantly. -
Key words:
- Uricase deficiency /
- Kunming-DY rats /
- Hyperuricemia /
- Intestinal flora /
- Lactobacillus
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图 1 KDY大鼠的血尿酸水平和体重变化[（ $\bar x \pm s $），n = 6]
A：45 d龄大鼠血清尿酸水平；B：45 d龄大鼠体重（$\bar x \pm s $，n = 6）。WT：野生型SD大鼠；Uox-/-：KDY大鼠。*P < 0.05 （双尾）。
Figure 1. Changes of blood uric acid level and body weight in KDY rats[（ $\bar x \pm s $），n = 6]
图 2 KDY大鼠各肠段菌群丰度变化[（ $ \bar x $），n = 6]
A：同龄大鼠门水平；B：同龄大鼠纲水平；C：同龄大鼠目水平；D：同龄大鼠科水平；E：同龄大鼠属水平（均数，n = 3）。WT：野生型SD大鼠；Uox-/-：KDY大鼠。其中1～18为小肠，19～20为结肠。
Figure 2. Variation of flora abundance in each intestinal segment of KDY rats[（ $\bar x $），n = 6]
图 3 KDY大鼠各节段菌群属水平多样性变化
与野生型大鼠（WT）相比，KDY大鼠（Uox-/-）各节段菌群属水平多样性变化（$\bar x \pm s $，n = 3）。其中1～18为小肠，19～20为结肠。独立分组t检验，*P < 0.05 （双尾）。
Figure 3. Variation of microflora at genus level in each segment of KDY rats
图 4 KDY大鼠新鲜粪便菌群丰度变化
与野生型（WT）大鼠相比，KDY（Uox-/-）大鼠新鲜粪便菌群丰度变化（均数，n = 6）。A：大鼠新鲜粪便菌群丰度在门水平变化；B：大鼠新鲜粪便菌群丰度在纲水平变化；C：大鼠新鲜粪便菌群丰度在目水平变化；D：大鼠新鲜粪便菌群丰度在科水平变化；E：大鼠新鲜粪便菌群丰度在属水平变化。*P < 0.05 （双尾）。
Figure 4. Abundance changes of fresh fecal flora in KDY rats
图 5 KDY大鼠新鲜粪便不同水平的种类差异
与野生型大鼠（WT）相比，KDY大鼠（Uox-/-）新鲜粪便不同水平的种类差异（$\bar x \pm s $，n = 6）。
Figure 5. Species differences of fresh feces in KDY rats at different levels
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[1] 林淑芃. 《中国高尿酸血症与痛风诊疗指南(2019)》解读[J]. 临床内科杂志,2020,37(6):460-462. doi: 10.3969/j.issn.1001-9057.2020.06.022 [2] 张楠,胡欣瑜,董鲜祥,等. 高尿酸血症动物模型的研究进展[J]. 昆明医科大学学报,2019,40(6):129-134. doi: 10.3969/j.issn.1003-4706.2019.06.027 [3] Lu J,Hou X,Yuan X,et al. Knockout of the urate oxidase gene provides a stable mouse model of hyperuricemia associated with metabolic disorders[J]. Kidney Int,2018,93(1):69-80. doi: 10.1016/j.kint.2017.04.031 [4] Yu Y,Zhang N,Dong X,et al. Uricase-deficient rat is generated with CRISPR/Cas9 technique[J]. Peer J,2020,8:e8971. doi: 10.7717/peerj.8971 [5] Sun X,Wen J,Guan B,et al. Folic acid and zinc improve hyperuricemia by altering the gut microbiota of rats with high-purine diet-induced hyperuricemia[J]. Front Microbiol,2022,13:907952. doi: 10.3389/fmicb.2022.907952 [6] 朱发伟,楼招欢. 桑抹茶对高尿酸血症模型大鼠血尿酸水平及肠道菌群的影响[J]. 中国现代应用药学,2017,34(8):1084-1088. doi: 10.13748/j.cnki.issn1007-7693.2017.08.003 [7] 殷娜,李小丝,刘维超,等. 健脾渗湿方对尿酸酶基因缺失高尿酸血症模式动物肾损伤及肠道微生态干预的研究[J]. 云南中医学院学报,2021,44(4):9-16. doi: 10.19288/j.cnki.issn.1000-2723.2021.04.002 [8] 董鲜祥,范楠,云宇,等. 排毒养颜胶囊对尿酸酶缺失大鼠的降血尿酸作用[J]. 云南中医学院学报,2020,43(3):1-4. doi: 10.19288/j.cnki.issn.1000-2723.2020.03.001 [9] 綦雅琳,秦婉,杲银芳,等. 75%摄食量对尿酸酶缺失大鼠的降尿酸作用[J]. 云南中医学院学报,2021,44(5):7-12. [10] 秦婉,綦雅琳,杲银芳,等. 聚乙二醇4000对Kunming-DY大鼠的降尿酸作用[J]. 昆明医科大学学报,2022,43(8):1-6. [11] Yin N,Li X,Liu W,et al. Jian Pi Shen Shi formula alleviates hyperuricemia and related renal fibrosis in uricase-deficient rats via suppression of the collagen-binding pathway[J]. Int J Rheum Dis,2022,25(12):1395-1407. [12] Gao Y,Yu Y,Qin W,et al. Uricase-deficient rats with similarly stable serum uric acid to human's are sensitive model animals for studying hyperuricemia[J]. PLoS One,2022,17(3):e0264696. doi: 10.1371/journal.pone.0264696 [13] Fan N,Li L,Xia H,et al. Aboriginal Bacterial Flora in the Uricase-Deficient Rat Gut is Not the Main Factor Affecting Serum Uric Acid[J]. Evid Based Complement Alternat Med,2021,2021:5587642. [14] 赵晖,陈军梅,盛冬云. 四妙散加味对高尿酸血症大鼠尿酸代谢及肠道菌群的影响[J]. 湖北中医药大学学报,2022,24(1):26-30. doi: 10.3969/j.issn.1008-987x.2022.01.06 [15] Fan N,Yu Y,Li L,et al. Uricase deficiency causes mild and multiple organ injuries in rats[J]. PLoS One,2021,16(8):e0256594. doi: 10.1371/journal.pone.0256594 -
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