犬慢性肾衰竭进程中肠道菌群代谢物短链脂肪酸水平的变化及其对肾功能的影响

doi:10.11843/j.issn.0366-6964.2021.08.027

摘要/Abstract

摘要： 本研究旨在评估短链脂肪酸在慢性肾衰竭患犬和健康犬中的水平，探究短链脂肪酸变化的原因及其对肾功能的影响。选取22例轻度慢性肾衰患犬（M-CRF组）、29例重度慢性肾衰患犬（S-CRF组）和26例健康对照犬（HC组），用16S rDNA测序技术分析肠道菌群多样性，气相色谱法检测粪中短链脂肪酸浓度。通过粪菌移植和补充丁酸钠给5/6肾摘除犬，观察肠道菌群及丁酸钠对肾功能影响。结果显示：1）S-CRF组肠道菌群多样性指标观察物种数及Simpson指数低于HC组（P<0.05），PCoA分析显示，S-CRF组肠道菌群与M-CRF、HC组有差异。2）LEfSe分析显示，S-CRF组和HC组间大量差异菌群，拟杆菌科、拟杆菌属及假单胞菌科等7个菌种富集于S-CRF组，普氏杆菌科、梭菌科、普氏杆菌属及普拉梭菌属等11个菌种富集于HC组。CCA分析发现富集于S-CRF组菌种丰度与肾功能指标呈正相关。3）S-CRF组粪中乙酸、丙酸及丁酸浓度均显著低于HC组和M-CRF组，M-CRF组丁酸浓度显著低于HC组（P<0.05），且丁酸浓度与血中胱抑素C（Cys-c）、肌酐（Cr）及尿素氮（BUN）等肾功能指标呈负相关（r值分别为-0.451、-0.583和-0.514，P<0.01）。4）与慢性肾衰模型组（5/6 Nx组）比较，慢性肾衰犬给与丁酸钠8周后，血清Cr和BUN显著降低（P<0.05）；粪菌移植8周后，血清Cr和BUN显著升高（P<0.05），丁酸钠可回调血清Cr和BUN水平。综上表明，慢性肾衰竭患犬肠道菌群多样性降低，菌群结构及丰度改变，粪中短链脂肪酸浓度降低，这些变化可加剧肾功能障碍。为犬慢性肾衰竭的防治提供新的理论依据。

关键词: 慢性肾衰竭, 犬, 肠道菌群, 短链脂肪酸, 肾功能

Abstract: This study aimed to evaluate the levels of short chain fatty acids in dogs with chronic renal failure and healthy dogs, and explore the causes of changes in short chain fatty acids and its impact on renal function. Twenty-two dogs with mild chronic renal failure (M-CRF group), 29 dogs with severe chronic renal failure (S-CRF group) and 26 healthy dogs (HC group) were selected. The diversity of gut microbiota was analyzed by 16S rDNA sequencing technology and the concentration of short chain fatty acids in feces was detected by gas chromatography. The effects of gut microbiota and sodium butyrate on renal function of 5/6 nephrectomized dogs were observed by fecal bacteria transplantation and sodium butyrate supplementation. The results showed that:1) the observed species and Simpson index of gut microflora diversity in S-CRF group were lower than those in HC group (P<0.05). PCoA analysis showed that the gut microflora in S-CRF group was different from that both in M-CRF and HC group. 2) Lefse analysis showed that there were a large number of different flora between S-CRF group and HC group. Seven species (bacteroideae, Bacteroides, Pseudomonas, et al) were enriched in S-CRF group, while eleven species, such as Prevotellaceae, Clostridiaceae, Prevotella, Faecalibacterium and so on, were enriched in HC group. CCA analysis showed that the species richness in S-CRF group and HC group were positively correlated and negatively correlated with renal function indexes, respectively. 3) The fecal concentrations of acetic acid, propionic acid and butyric acid in S-CRF group were significantly lower than those in HC group and M-CRFgroup, and butyric acid concentration in M-CRF group was significantly lower than that in HC group too (P<0.05). Furthermore, butyric acid concentration was negatively correlated with serum cystatin C (Cys-C), creatinine (Cr) and urea nitrogen (BUN),and the correlation coefficients were -0.451, -0.583 and -0.514 (P<0.01), respectively. 4) Compared with chronic renal failure model group (5/6 Nx group), the serum Cr and BUN level of 5/6 nephrectomized dog fed sodium butyrate were significantly decreased after 8 weeks (P<0.05). Serum Cr and BUN level of 5/6 nephrectomized mice transplanted fecal bacteria from dogs with CRF were more higher than those in 5/6 Nx group after 8 weeks (P<0.05), while sodium butyrate could reverse these changes. In summary, chronic renal failure of dogs can lead to the decrease of gut microflora diversity, the change of flora structure and abundance, and the decrease of fecal short chain fatty acids concentration. It provides a new theoretical basis for the prevention and treatment of chronic renal failure in dogs.

Key words: chronic renal failure, dog, gut microbiota, short-chain fatty acids, renal function

中图分类号:

S856.59

刘静, 朱道仙, 卢劲晔, 张一多, 卢炜, 陆江. 犬慢性肾衰竭进程中肠道菌群代谢物短链脂肪酸水平的变化及其对肾功能的影响[J]. 畜牧兽医学报, 2021, 52(8): 2334-2343.

LIU Jing, ZHU Daoxian, LU Jinye, ZHANG Yiduo, LU Wei, LU Jiang. Alteration of Short-Chain Fatty Acids Produced by Gut Microflora in Dogs with Chronic Renal Failure and Its Effect on Renal Function[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(8): 2334-2343.

参考文献 19

[1]	肖华平, 黄忍, 任桥. 犬猫慢性肾衰竭综述[J]. 山东畜牧兽医, 2019, 40(5):72-73.XIAO H P, HUANG R, REN Q. Review of chronic renal failure in dogs and cats[J]. Shandong Journal of Animal Science and Veterinary Medicine, 2019, 40(5):72-73.(in Chinese)
[2]	ROWLAND I, GIBSON G, HEINKEN A, et al. Gut microbiota functions:metabolism of nutrients and other food components[J]. Eur J Nutr, 2018, 57(1):1-24.
[3]	SWARTWOUT B, LUO X M. Implications of probiotics on the maternal-neonatal interface:gut microbiota, immunomodulation, and autoimmunity[J]. Front Immunol, 2018, 9:2840.
[4]	CORNEJO-PAREJA I, MUÑOZ-GARACH A, CLEMENTE-POSTIGO M, et al. Importance of gut microbiota in obesity[J]. Eur J Clin Nutr, 2019, 72(Suppl 1):26-37.
[5]	LI W Z, STIRLING K, YANG J J, et al. Gut microbiota and diabetes:from correlation to causality and mechanism[J]. World J Diabetes, 2020, 11(7):293-308.
[6]	CAPORASO J G, KUCZYNSKI J, STOMBAUGH J, et al. QIIME allows analysis of high-throughput community sequencing data[J]. Nat Methods, 2010, 7(5):335-336.
[7]	孙翠丽, 张阁, 程汝佳, 等. 16S rRNA高通量测序方法检测羊圈空气微生物群落结构及多样性[J]. 畜牧兽医学报, 2017, 48(7):1314-1322.SUN C L, ZHANG G, CHENG R J, et al. Microbial community structure and diversity of sheepfold atmosphere by 16S rRNA high-throughput sequencing[J]. Acta Veterinaria Et Zootechnica Sinica, 2017, 48(7):1314-1322. (in Chinese)
[8]	徐帅, 林奕岑, 周梦佳, 等. 基于高通量测定肉鸡回肠微生物多样性及PICRUSt基因预测分析[J]. 动物营养学报, 2016, 28(8):2581-2588.XU S, LIN Y C, ZHOU M J, et al. Analyzing of ileum microbial diversity of chickens by high-throughput sequencing and PICRUSt predicted[J]. Chinese Journal of Animal Nutrition, 2016, 28(8):2581-2588.(in Chinese)
[9]	刘珍妮, 夏霆, 孙淑文, 等. 苏北运东水网区浮游植物群落结构特征与环境因子的关系[J]. 水生态学杂志, 2019, 40(6):45-53.LIU Z N, XIA T, SUN S W, et al. Phytoplankton community structure and its relationship with environment factors in the Yundong River network of northern Jiangsu[J]. Journal of Hydroecology, 2019, 40(6):45-53.(in Chinese)
[10]	ZHAO L P, ZHANG F, DING X Y, et al. Gut bacteria selectively promoted by dietary fibers alleviate type 2 diabetes[J]. Science, 2018, 359(6380):1151-1156.
[11]	王宁, 李宁, 陈洋. 肾动脉结扎法制作慢性肾功能衰竭犬模型方法的探索[J]. 中国医科大学学报, 2006, 35(2):221.WANG N, LI N, CHEN Y. Chronic renal failure model in canines by ligation of the renal artery branches[J]. Journal of China Medical University, 2006, 35(2):221.(in Chinese)
[12]	BANERJEE S, SAR A, MISRA A, et al. Increased productivity in poultry birds by sub-lethal dose of antibiotics is arbitrated by selective enrichment of gut microbiota, particularly short-chain fatty acid producers[J]. Microbiology, 2018, 164(2):142-153.
[13]	ARUMUGAM M, RAES J, PELLETIER E, et al. Enterotypes of the human gut microbiome[J]. Nature, 2011, 473(7346):174-180.
[14]	WONG J, PICENO Y M, DESANTIS T Z, et al. Expansion of urease- and uricase-containing, indole- and p-cresol-forming and contraction of short-chain fatty acid-producing intestinal microbiota in ESRD[J]. Am J Nephrol, 2014, 39(3):230-237.
[15]	LIU H, WANG J, HE T, et al. Butyrate:a double-edged sword for health?[J]. Adv Nutr, 2018, 9(1):21-29.
[16]	SCHULTHESS J, PANDEY S, CAPITANI M, et al. The short chain fatty acid butyrate imprints an antimicrobial program in macrophages[J]. Immunity, 2019, 50(2):432-445.
[17]	LEE S U, IN H J, KWON M S, et al. β-Arrestin 2 mediates G protein-coupled receptor 43 signals to nuclear factor-κB[J]. Biol Pharm Bull, 2013, 36(11):1754-1759.
[18]	NOURLDEIN M H, EID A A. Gut microbiota and mTOR signaling:insight on a new pathophysiological interaction[J]. Microb Pathog, 2018, 118:98-104.
[19]	高冰, 于秋颖, 贾海莲, 等. 短链脂肪酸提取与检测的研究进展[J]. 包头医学院学报, 2019, 35(12):116-117.GAO B, YU Q Y, JIA H L, et al. Research progress on extraction and detection of short chain fatty acids[J]. Journal of Baotou Medical College, 2019, 35(12):116-117.(in Chinese)