实验方法与实验动物 EXPERIMENTAL METHOD AND ANIMAL

D-天冬氨酸对葡聚糖硫酸钠诱导结肠炎小鼠抗氧化和炎症反应的影响

  • 胡鲜 ,
  • 王晨昱 ,
  • 刘娣 ,
  • 龙静 ,
  • 范文君 ,
  • 何流琴 ,
  • 李铁军 ,
  • 姚继明
展开
  • 1. 湖南师范大学生命科学学院, 动物肠道功能与调控湖南省重点实验室, 长沙 410081;
    2. 中国科学院亚热带农业生态研究所, 畜禽养殖污染控制与资源化技术国家工程实验室, 动物营养生理与代谢过程湖南省重点实验室, 长沙 410125;
    3. 中国科学院大学, 北京 100049;
    4. 黑龙江省农业科学院, 哈尔滨 150086;
    5. 广东旺大集团股份有限公司, 广州 510663
胡鲜(1996-),女,贵州普安人,硕士研究生,从事动物营养研究。E-mail:1654027745@qq.com

收稿日期: 2021-01-29

  网络出版日期: 2021-08-11

基金资助

国家自然科学基金项目(31902168,31872371);中国科协青年托举人才项目(2019QNRC001);广西桂林技术创新引导计划(2020010901);广西重点研发计划(桂科AB19259012);长沙市科技计划项目(kq1907073)

Effects of D-Aspartic on Antioxidant and Inflammation of Dextran Sodium Sulfate Induced Colitis of Mice

  • HU Xian ,
  • WANG Chenyu ,
  • LIU Di ,
  • LONG Jing ,
  • FAN Wenjun ,
  • HE Liuqin ,
  • LI Tiejun ,
  • YAO Jiming
Expand
  • 1. Hunan Province Key Laboratory of Animal Intestinal Function and Regulation, College of Life Sciences, Hunan Normal University, Changsha 410081, China;
    2. Hunan Provincial Key Laboratory of Animal Nutrition Physiology and Metabolic Processes, National Engineering Laboratory of Poultry Breeding Pollution Control and Resource Technology, Institute of Subtropical Agricultural Ecology, Chinese Academy of Sciences, Changsha 410125, China;
    3. University of Chinese Academy of Sciences, Beijing 100049, China;
    4. Heilongjiang Academy of Agricultural Sciences, Harbin 150086, China;
    5. Guangdong Wangda Group Co., Ltd., Guangzhou 510663, China

Received date: 2021-01-29

  Online published: 2021-08-11

Supported by

 

摘要

本试验旨在探讨D-天冬氨酸(D-Asp)对葡聚糖硫酸钠(DSS)诱导的小鼠结肠炎肠道损伤的恢复效果。试验选取40只体重相近的C57BL/6J雄性小鼠,随机分为5组,即对照组、0 mmol/L D-Asp组、0.15 mmol/L D-Asp组、2.30 mmol/L D-Asp组和7.70 mmol/L D-Asp组,每组8个重复,每个重复1只,单笼饲养。试验第1~7天,对照组小鼠正常饮用蒸馏水,其他各组小鼠均饮用含3.5% DSS的蒸馏水,诱导小鼠结肠炎。试验第8天开始,对照组和0 mmol/L D-Asp组小鼠直肠灌注0.1 mL生理盐水,其他各组分别直肠灌注0.1 mL浓度为0.15、2.30和7.70 mmol/L的D-Asp溶液,每隔1 d灌注1次,共灌注5次。结果表明:1)与对照组相比,0 mmol/L D-Asp组小鼠第5天体重和平均日采食量显著降低(P<0.05),结肠长度变短并出现浸血现象。与0 mmol/L D-Asp组相比,7.70 mmol/L D-Asp组小鼠第12~14天体重和平均日采食量显著提高(P<0.05),结肠变长,绒毛高度增高及隐窝深度变浅,杯状细胞排列更规则。2)与0 mmol/L D-Asp组相比,7.70 mmol/L D-Asp组的血清白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)含量和嗜酸性粒细胞过氧化物酶(EPO)活性显著降低(P<0.05),结肠中天冬氨酸/谷氨酸载体2(SLC25A13) mRNA相对表达量显著升高(P<0.05),结肠中白细胞介素-6(IL-6)、乙酰辅酶A羧化酶(ACC)和天冬氨酸/谷氨酸载体1(SLC25A12) mRNA相对表达量显著降低(P<0.05)。由此可见,直肠灌注D-Asp可通过提高抗氧化酶的活性和相关基因的表达,抑制炎性细胞因子的分泌,改善结肠形态结构,从而促进因DSS引起的小鼠结肠炎损伤修复。

本文引用格式

胡鲜 , 王晨昱 , 刘娣 , 龙静 , 范文君 , 何流琴 , 李铁军 , 姚继明 . D-天冬氨酸对葡聚糖硫酸钠诱导结肠炎小鼠抗氧化和炎症反应的影响[J]. 动物营养学报, 2021 , 33(8) : 4708 -4718 . DOI: 10.3969/j.issn.1006-267x.2021.08.049

Abstract

This experiment was conducted to study the repairing effects of D-aspartic acid (D-Asp) on dextran sodium sulfate (DSS) induced colonic intestinal injury of mice. A total of 40 C57BL/6J male mice with similar body weight were randomly divided into 5 groups, which were the control group, 0 mmol/L D-Asp group, 0.15 mmol/L D-Asp group, 2.30 mmol/L D-Asp group and 7.70 mmol/L D-Asp group, each group contained 8 replicates and each replicate contained 1 mouse, the mice were reared in a single cage. During days 1 to 7, the mice in the control group drank sterile water, while the others in other groups drank sterile water containing 3.5% DSS, to induce colitis of mice. From day 8, the control group and 0 mmol/L D-Asp group were rectal perfused with 0.1 mL normal saline, and the mice in the other groups were rectal perfused with 0.1 mL D-Asp at the concentrations of 0.15, 2.30 and 7.70 mmol/L, respectively, and perfused once every 1 d for 5 times. The results showed as follows:1) compared with the control group, the body weight on day 5 and average daily feed intake of 0 mmol/L D-Asp group were significantly decreased (P<0.05), and the colon length was shortened and the phenomenon of blood infiltration occurred. Compared with the 0 mmol/L D-Asp group, the body weight on days 12 to 14 and average daily feed intake of 7.70 mmol/L D-Asp group were significantly increased (P<0.05), the colon became longer, the villi height and crypt depth became shallower, and the arrangement of goblet cells was more regular. 2) Compared with the 0 mmol/L D-Asp group, the serum interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) contents and eosinophil peroxidase (EPO) activity of 7.70 mmol/L D-Asp group were significantly decreased (P<0.05), the mRNA relative expression level of aspartate/glutamate carrier 2 (SLC25A13) in colon was significantly increased (P<0.05), and the mRNA relative expression levels of interleukin-6 (IL-6), acetyl CoA carboxylase (ACC) and aspartate/glutamate carrier 1 (SLC25A12) in colon were significantly decreased (P<0.05). In conclusion, rectal perfused D-Asp can inhibit the secretion of inflammatory cytokines and improve the morphological structure of colon by increasing the activity of antioxidant enzymes and the expression of related genes, thereby promoting the repair of mouse colitis damage caused by DSS.

参考文献

[1] ZHANG Q,XU N N,HU X J,et al.Anti-colitic effects of physalin B on dextran sodium sulfate-induced balb/c mice by suppressing multiple inflammatory signaling pathways[J].Journal of Ethnopharmacology,2020,259:112956.
[2] SHENG K L,ZHANG G H,SUN M,et al.Grape seed proanthocyanidin extract ameliorates dextran sulfate sodium-induced colitis through intestinal barrier improvement,oxidative stress reduction,and inflammatory cytokines and gut microbiota modulation[J].Food & Function,2020,11(9):7817-7829.  
[3] 韩伟,刘宇琼,孙雷.丹参、黄芪注射液治疗溃疡性结肠炎的临床研究[J].长春中医学院学报,2005,21(4):7. HAN W,LIU Y Q,SUN L.Clinical study of Danshen and Huangqi injection in the treatment of ulcerative colitis[J].Journal of Changchun College of Traditional Chinese Medicine,2005,21(4):7.(in Chinese)
[4] 蔡彬,周敏红,李春伟.氨基酸在炎症性肠病治疗中的研究进展[J].中国医药导报,2020,17(9):37-40. CAI B,ZHOU M H,LI C W.Research progress of amino acids in the treatment of inflammatory bowel disease[J].China Medical Herald,2020,17(9):37-40.(in Chinese)
[5] 夏嗣廷,胡睿智,贺建华,等.D-天冬氨酸生理功能及其在畜禽生产中的应用[J].动物营养学报,2019,31(10):4481-4487. XIA S T,HU R Z,HE J H,et al.Physiological function of D-aspartic acid and its application in livestock and poultry production[J].Chinese Journal of Animal Nutrition,2019,31(10):4481-4487.(in Chinese)
[6] FENG Z Q Q,XU B.Inspiration from the mirror:D-amino acid containing peptides in biomedical approaches[J].Biomolecular Concepts,2016,7(3):179-187.  
[7] SASABE J,SUZUKI M.Emerging role of D-amino acid metabolism in the innate defense[J].Frontiers in Microbiology,2018,9:933.
[8] WU G Y.Intestinal mucosal amino acid catabolism[J].The Journal of Nutrition,1998,128(8):1249-1252.  
[9] ZHANG H W,HUA R,ZHANG B X,et al.Serine alleviates dextran sulfate sodium-induced colitis and regulates the gut microbiota in mice[J].Frontiers in Microbiology,2018,9:3062.
[10] BOCCELLA S,VACCA V,ERRICO F,et al.D-aspartate modulates nociceptive-specific neuron activity and pain threshold in inflammatory and neuropathic pain condition in mice[J].BioMed Research International,2015,2015:905906.
[11] D'ANIELLO A.D-aspartic acid:an endogenous amino acid with an important neuroendocrine role[J].Brain Research Reviews,2007,53(2):215-234.  
[12] ERWAN E,CHOWDHURY V S,NAGASAWA M,et al.Oral administration of D-aspartate,but not L-aspartate,depresses rectal temperature and alters plasma metabolites in chicks[J].Life Sciences,2014,109(1):65-71.  
[13] 段杰林.酸性氨基酸缓解过氧化氢介导仔猪肠道氧化损伤机制研究[D].硕士学位论文.北京:中国科学院大学,2016. DUAN J L.Study on the mechanism of acidic amino acids alleviating hydrogen peroxide-mediated intestinal oxidative damage in piglets[D].Master's Thesis.Beijing:University of Chinese Academy of Sciences,2016.(in Chinese)
[14] YOUSEF M,PICHYANGKURA R,SOODVILAI S,et al.Chitosan oligosaccharide as potential therapy of inflammatory bowel disease:therapeutic efficacy and possible mechanisms of action[J].Pharmacological Research,2012,66(1):66-79.  
[15] CHAO L,ZHENG P Y,XIA L,et al.Calycosin attenuates dextran sulfate sodium (DSS)-induced experimental colitis[J].Journal of Basic Medical Sciences,2017,20(9):1056-1062.
[16] LIN X C,YI Z Y,DIAO J X,et al.Shaoyao decoction ameliorates colitis-associated colorectal cancer by downregulating proinflammatory cytokines and promoting epithelial-mesenchymal transition[J].Journal of Translational Medicine,2014,12:105.
[17] POPIVANOVA B K,KITAMURA K,WU Y,et al.Blocking TNF-alpha in mice reduces colorectal carcinogenesis associated with chronic colitis[J].Journal of Clinical Investigation,2008,118(2):560-570.
[18] ERWAN E,TOMONAGA S,YOSHIDA J,et al.Central administration of L- and D-aspartate attenuates stress behaviors by social isolation and crf in neonatal chicks[J].Amino Acids,2012,43(5):1969-1976.  
[19] LI Y Y,HAN H,YIN J,et al.D- and L-aspartate regulates growth performance,inflammation and intestinal microbial community in young pigs[J].Food & Function,2019,10(2):1028-1037.  
[20] CABISCOL E,TAMARIT J,ROS J.Oxidative stress in bacteria and protein damage by reactive oxygen species[J].International Microbiology,2000,3(1):3-8.
[21] TANG W J,WU J,JIN S S,et al.Glutamate and aspartate alleviate testicular/epididymal oxidative stress by supporting antioxidant enzymes and immune defense systems in boars[J].Science China Life Sciences,2020,63(1):116-124.  
[22] BARBATO V,TALEVI R,BRAUN S,et al.Supplementation of sperm media with zinc,D-aspartate and co-enzyme q10 protects bull sperm against exogenous oxidative stress and improves their ability to support embryo development[J].Zygote,2017,25(2):168-175.  
[23] 郑贵花,夏广军,尹宝珍.乙酰辅酶A羧化酶在动物中的研究进展[J].现代农业研究2019(10):109-112,120. ZHENG G H,XIA G J,YIN B Z.Research progress of acetyl-coenzyme A carboxylase in animals[J].Modern Agricultural Research,2019(10):109-112,120.(in Chinese)
[24] 谢宝娟,向霖丽,许文娟,等.乙酰辅酶A羧化酶激动剂柠檬酸钠对慢性哮喘小鼠气道炎症和气道重塑的作用[J].武汉大学学报(医学版),2021,42(2):267-271. XIE B J,XIANG L L,XU W J.et al.Effects of acetyl-coenzyme A carboxylase agonist sodium citrate on airway inflammation and airway remodeling in mice with chronic asthma[J].Journal of Wuhan University (Medical Edition),2021,42(2):267-271.(in Chinese)
[25] 黄伟聪.髓过氧化物酶、氧化应激与多囊卵巢综合征相关性研究进展[J].现代医药生物,2020,36(5):82-85. HUANG W C.Research progress on the correlation between myeloperoxidase,oxidative stress and polycystic ovary syndrome[J].Modern Medical Biology,2020,36(5):82-85.(in Chinese)
[26] 刘顺爱,杨忠贞.嗜酸性粒细胞与组织免疫损伤[J].国外医学(免疫学分册),1993,16(5):235-239. LIU S A,YANG Z Z.Eosinophils and tissue immune damage[J].Foreign Medicine (Immunology),1993,16(5):235-239.(in Chinese)
[27] AMOEDO N D,PUNZI G,OBRE E,et al.Agc1/2,the mitochondrial aspartate-glutamate carriers[J].Biochimica et Biophysica Acta:Molecular Cell Research,2016,1863(10):2394-2412.  
[28] PUERTAS-FRIAS G,DEL ARCO A,PARDO B,et al.Mitochondrial movement in aralar/Slc25a12/AGC1 deficient cortical neurons[J].Neurochemistry International,2019,131:104541.
[29] PALAZZO E,LUONGO L,GUIDA F,et al.D-aspartate drinking solution alleviates pain and cognitive impairment in neuropathic mice[J].Amino Acids,2016,48(7):1553-1567.  
文章导航

/