Amino Acids

pp 1–11 | Cite as

Glycine supplementation to breast-fed piglets attenuates post-weaning jejunal epithelial apoptosis: a functional role of CHOP signaling

  • Xiaoxiao Fan
  • Shuai Li
  • Zhenlong WuEmail author
  • Zhaolai Dai
  • Ju Li
  • Xiaolong Wang
  • Guoyao Wu
Original Article


This study was conducted to test the hypothesis that preweaning  glycine supplementation to breast-fed piglets alleviated the post-weaning  apoptosis of jejunal epithelium through CHOP signaling. Seven-day-old sow-reared piglets were orally administrated with 0, 50, 100, or 200% of glycine intake from sow’s milk twice daily for 14 days and then were weaned at 21 days of age. Tissue samples were collected at 28 days of age for determining intestinal morphology, serum diamine oxidase (DAO) activity, abundances of proteins involved in ER stress and apoptosis. Glycine (100–200%) administration increased villus height, the ratio of villus height to crypt depth in the jejunum. Glycine supplementation (200%) enhanced average daily weight gain during the first 2 weeks post-weaning. Serum DAO activity and jejunal epithelium apoptosis were decreased, but the number of goblet cells in the jejunum was increased. Western blot analysis showed that 100–200% glycine enhanced the protein levels of occludin, claudin-1, and zonula occludens (ZO)-1 without affecting those of claudin-3, ZO-2, and ZO-3. Further studies showed that protein abundances of glucose-regulated protein 78 (BiP/GRP78) and p-IRE1α, instead of ATF6α, were reduced by glycine. Among the proteins related to apoptosis, abundances of CHOP and p53 were reduced, whereas those of Bcl-2 and Bcl-xL were enhanced in the jejunum of 100–200% glycine-supplemented piglets. Collectively, our results indicated that preweaning glycine supplementation improved the intestinal development of post-weaning piglets. The beneficial effect of glycine was associated with improved intestinal mucosal barrier and reduced apoptosis of enterocytes through CHOP signaling.


Glycine Endoplasmic reticulum stress Small intestine Apoptosis Piglets 



This work was supported by the National Natural Science Foundation of China (no. 31572412, 31572410, 31625025, 31272451 and  31272450), the Zhengzhou 1125 Talent Program, Agriculture and Food Research Initiative Competitive Grants (2014-67015-21770, 2015-67015-23276 and 2016-67015-24958) from the USDA National Institute of Food and Agriculture, and Texas A&M AgriLife Research (H-8200).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethics statement

The studies were approved by China Agricultural University Institutional Animal Science and Technology College and conducted according to the Guidelines for Experimental Animal Research of the Ministry of Science and Technology (Beijing, China).

Informed consent

All authors have read and approved the final manuscript.

Supplementary material

726_2018_2681_MOESM1_ESM.docx (13 kb)
Supplementary material 1 (DOCX 14 KB)


  1. Amelio I, Cutruzzola F, Antonov A, Agostini M, Melino G (2014) Serine and glycine metabolism in cancer. Trends Biochem Sci 39(4):191–198CrossRefGoogle Scholar
  2. Amin K, Li J, Chao WR, Dewhirst MW, Haroon ZA (2003) Dietary glycine inhibits angiogenesis during wound healing and tumor growth. Cancer Biol Ther 2(2):173–178CrossRefGoogle Scholar
  3. Arrieta MC, Bistritz L, Meddings JB (2006) Alterations in intestinal permeability. Gut 55(10):1512–1520PubMedGoogle Scholar
  4. Bauer E, Metzler-Zebeli BU, Verstegen MW, Mosenthin R (2011) Intestinal gene expression in pigs: effects of reduced feed intake during weaning and potential impact of dietary components. Nutr Res Rev 24(2):155–175CrossRefGoogle Scholar
  5. Bertolotti A, Zhang Y, Hendershot LM, Harding HP, Ron D (2000) Dynamic interaction of BiP and ER stress transducers in the unfolded-protein response. Nat Cell Biol 2(6):326–332CrossRefGoogle Scholar
  6. Bhattacharyya S, Ghosh J, Sil PC (2012) Iron induces hepatocyte death via MAPK activation and mitochondria-dependent apoptotic pathway: beneficial role of glycine. Free Radic Res 46(10):1296–1307CrossRefGoogle Scholar
  7. Birchenough GM, Johansson ME, Gustafsson JK, Bergstrom JH, Hansson GC (2015) New developments in goblet cell mucus secretion and function. Mucosal Immunol 8(4):712–719CrossRefGoogle Scholar
  8. Campbell JM, Crenshaw JD, Polo J (2013) The biological stress of early weaned piglets. J Anim Sci Biotechno 4:19CrossRefGoogle Scholar
  9. Chen JQ, Ma XS, Yang Y, Dai ZL, Wu ZL, Wu G (2018) Glycine enhances expression of adiponectin and IL-10 in 3T3-L1 adipocytes without affecting adipogenesis and lipolysis. Amino Acids 50:629–640CrossRefGoogle Scholar
  10. Dai Z, Wu Z, Hang S, Zhu W, Wu G (2015) Amino acid metabolism in intestinal bacteria and its potential implications for mammalian reproduction. Mol Hum Reprod 21(5):389–409CrossRefGoogle Scholar
  11. de Aguiar Picanco E, Lopes-Paulo F, Marques RG, Diestel CF, Caetano CE, de Souza MV, Moscoso GM, Pazos HM (2011) L-arginine and glycine supplementation in the repair of the irradiated colonic wall of rats. Int J Colorectal Dis 26(5):561–568CrossRefGoogle Scholar
  12. Diestel CF, Marques RG, Lopes-Paulo F, Paiva D, Horst NL, Caetano CE, Portela MC (2007) Role of l-glutamine and glycine supplementation on irradiated colonic wall. Int J Colorectal Dis 22(12):1523–1529CrossRefGoogle Scholar
  13. Fuchs SA, Peeters-Scholte CM, de Barse MM, Roeleveld MW, Klomp LW, Berger R, de Koning TJ (2012) Increased concentrations of both NMDA receptor co-agonists d-serine and glycine in global ischemia: a potential novel treatment target for perinatal asphyxia. Amino Acids 43(1):355–363CrossRefGoogle Scholar
  14. Gilani S, Howarth GS, Kitessa SM, Tran CD, Forder REA, Hughes RJ (2017) New biomarkers for increased intestinal permeability induced by dextran sodium sulphate and fasting in chickens. J Anim Physiol Anim Nutr (Berl) 101(5):e237–e245CrossRefGoogle Scholar
  15. Hall JC (1998) Glycine. J Parenter Enteral Nutr 22(6):393–398CrossRefGoogle Scholar
  16. Hetz C, Chevet E, Oakes SA (2015) Proteostasis control by the unfolded protein response. Nat Cell Biol 17(7):829–838CrossRefGoogle Scholar
  17. Hotamisligil GS (2010) Endoplasmic reticulum stress and the inflammatory basis of metabolic disease. Cell 140(6):900–917CrossRefGoogle Scholar
  18. Hou Y, Wu G (2018) L-Glutamate nutrition and metabolism in swine. Amino Acids 50(11):1497–1510CrossRefGoogle Scholar
  19. Hou Y, Yao K, Yin Y, Wu G (2016) Endogenous synthesis of amino acids limits growth, lactation, and reproduction in animals. Adv Nutr 7(2):331–342CrossRefGoogle Scholar
  20. Hu CH, Xiao K, Luan ZS, Song J (2013) Early weaning increases intestinal permeability, alters expression of cytokine and tight junction proteins, and activates mitogen-activated protein kinases in pigs. J Anim Sci 91(3):1094–1101CrossRefGoogle Scholar
  21. Iurlaro R, Munoz-Pinedo C (2016) Cell death induced by endoplasmic reticulum stress. FEBS J 283(14):2640–2652CrossRefGoogle Scholar
  22. Jacob T, Ascher E, Hingorani A, Kallakuri S (2003) Glycine prevents the induction of apoptosis attributed to mesenteric ischemia/reperfusion injury in a rat model. Surgery 134(3):457–466CrossRefGoogle Scholar
  23. Jacobi SK, Odle J (2012) Nutritional factors influencing intestinal health of the neonate. Adv Nutr 3(5):687–696CrossRefGoogle Scholar
  24. Kim SW, Wu G (2004) Dietary arginine supplementation enhances the growth of milk-fed young pigs. J Nutr 134(3):625–630CrossRefGoogle Scholar
  25. Kusche J, van Trotha U, Muhlberger G, Lorenz W (1974) The clinical-chemical application of the NADH test for the determination of diamine oxidase activity in human pregnancy. Agents Actions 4(3):188–189CrossRefGoogle Scholar
  26. Li P, Wu G (2018) Roles of dietary glycine, proline, and hydroxyproline in collagen synthesis and animal growth. Amino Acids 50:29–38CrossRefGoogle Scholar
  27. Li W, Sun K, Ji Y, Wu Z, Wang W, Dai Z, Wu G (2016) Glycine regulates expression and distribution of claudin-7 and ZO-3 Proteins in intestinal porcine epithelial cells. J Nutr 146(5):964–969CrossRefGoogle Scholar
  28. Lin WC, Chuang YC, Chang YS, Lai MD, Teng YN, Su IJ, Wang CC, Lee KH, Hung JH (2012) Endoplasmic reticulum stress stimulates p53 expression through NF-kappaB activation. PLoS One 7(7):e39120CrossRefGoogle Scholar
  29. Lu Y, Zhang J, Ma B, Li K, Li X, Bai H, Yang Q, Zhu X, Ben J, Chen Q (2012) Glycine attenuates cerebral ischemia/reperfusion injury by inhibiting neuronal apoptosis in mice. Neurochem Int 61(5):649–658CrossRefGoogle Scholar
  30. Marchiando AM, Graham WV, Turner JR (2010) Epithelial barriers in homeostasis and disease. Annu Rev Pathol 5:119–144CrossRefGoogle Scholar
  31. Melendez-Hevia E, De Paz-Lugo P, Cornish-Bowden A, Cardenas ML (2009) A weak link in metabolism: the metabolic capacity for glycine biosynthesis does not satisfy the need for collagen synthesis. J Biosci 34(6):853–872CrossRefGoogle Scholar
  32. Miyoshi J, Miyamoto H, Goji T, Taniguchi T, Tomonari T, Sogabe M, Kimura T, Kitamura S, Okamoto K, Fujino Y, Muguruma N, Okahisa T, Takayama T (2015) Serum diamine oxidase activity as a predictor of gastrointestinal toxicity and malnutrition due to anticancer drugs. J Gastroen Hepatol 30(11):1582–1590CrossRefGoogle Scholar
  33. Moens E, Veldhoen M (2012) Epithelial barrier biology: good fences make good neighbours. Immunology 135(1):1–8CrossRefGoogle Scholar
  34. Odenwald MA, Turner JR (2013) Intestinal permeability defects: is it time to treat? Clin Gastroenterol H 11(9):1075–1083CrossRefGoogle Scholar
  35. Ospina-Rojas IC, Murakami AE, Oliveira CA, Guerra AF (2013) Supplemental glycine and threonine effects on performance, intestinal mucosa development, and nutrient utilization of growing broiler chickens. Poult Sci 92(10):2724–2731CrossRefGoogle Scholar
  36. Petrat F, Drowatzky J, Boengler K, Finckh B, Schmitz KJ, Schulz R, de Groot H (2011) Protection from glycine at low doses in ischemia-reperfusion injury of the rat small intestine. Eur Surg Res 46(4):180–187CrossRefGoogle Scholar
  37. Powell S, Bidner TD, Payne RL, Southern LL (2011) Growth performance of 20- to 50-kg pigs fed low-crude-protein diets supplemented with histidine, cystine, glycine, glutamic acid, or arginine. J Anim Sci 89(11):3643–3650CrossRefGoogle Scholar
  38. Rhoads JM, Wu GY (2009) Glutamine, arginine, and leucine signaling in the intestine. Amino Acids 37(1):111–122CrossRefGoogle Scholar
  39. Ron D, Walter P (2007) Signal integration in the endoplasmic reticulum unfolded protein response. Nat Rev Mol Cell Biol 8(7):519–529CrossRefGoogle Scholar
  40. Sheng YH, Triyana S, Wang R, Das I, Gerloff K, Florin TH, Sutton P, McGuckin MA (2013) MUC1 and MUC13 differentially regulate epithelial inflammation in response to inflammatory and infectious stimuli. Mucosal Immunol 6(3):557–568CrossRefGoogle Scholar
  41. Stoffels B, Turler A, Schmidt J, Nazir A, Tsukamoto T, Moore BA, Schnurr C, Kalff JC, Bauer AJ (2011) Anti-inflammatory role of glycine in reducing rodent postoperative inflammatory ileus. Neurogastroenterol Motil 23(1):76–78CrossRefGoogle Scholar
  42. Sun Y, Wu Z, Li W, Zhang C, Sun K, Ji Y, Wang B, Jiao N, He B, Wang W, Dai Z, Wu G (2015) Dietary l-leucine supplementation enhances intestinal development in suckling piglets. Amino Acids 47(8):1517–1525CrossRefGoogle Scholar
  43. Sun KJ, Wu ZL, Ji Y, Wu G (2016) Glycine regulates protein turnover by activating protein kinase B/Mammalian target of rapamycin and by inhibiting MuRF1 and atrogin-1 gene expression in C2C12 myoblasts. J Nut 146:2461–2467CrossRefGoogle Scholar
  44. Szegezdi E, Logue SE, Gorman AM, Samali A (2006) Mediators of endoplasmic reticulum stress-induced apoptosis. EMBO Rep 7(9):880–885CrossRefGoogle Scholar
  45. Tossou MC, Liu H, Bai M, Chen S, Cai Y, Duraipandiyan V, Liu H, Adebowale TO, Al-Dhabi NA, Long L, Tarique H, Oso AO, Liu G, Yin Y (2016) Effect of high dietary tryptophan on intestinal morphology and tight junction protein of weaned pig. Biomed Res Int 2016:2912418CrossRefGoogle Scholar
  46. Turner JR (2009) Intestinal mucosal barrier function in health and disease. Nat Rev Immunol 9(11):799–809CrossRefGoogle Scholar
  47. Wang J, Wu Z, Li D, Li N, Dindot SV, Satterfield MC, Bazer FW, Wu G (2012) Nutrition, epigenetics, and metabolic syndrome. Antioxid Redox Signal 17(2):282–301CrossRefGoogle Scholar
  48. Wang W, Wu Z, Dai Z, Yang Y, Wang J, Wu G (2013) Glycine metabolism in animals and humans: implications for nutrition and health. Amino Acids 45(3):463–477CrossRefGoogle Scholar
  49. Wang W, Dai Z, Wu Z, Lin G, Jia S, Hu S, Dahanayaka S, Wu G (2014a) Glycine is a nutritionally essential amino acid for maximal growth of milk-fed young pigs. Amino Acids 46(8):2037–2045CrossRefGoogle Scholar
  50. Wang W, Wu Z, Lin G, Hu S, Wang B, Dai Z, Wu G (2014b) Glycine stimulates protein synthesis and inhibits oxidative stress in pig small intestinal epithelial cells. J Nutr 144(10):1540–1548CrossRefGoogle Scholar
  51. Wawryk-Gawda E, Chylinska-Wrzos P, Lis-Sochocka M, Chlapek K, Bulak K, Jedrych M, Jodlowska-Jedrych B (2014) P53 protein in proliferation, repair and apoptosis of cells. Protoplasma 251(3):525–533CrossRefGoogle Scholar
  52. Weinberg JM, Bienholz A, Venkatachalam MA (2016) The role of glycine in regulated cell death. Cell Mol Life Sci 73(11–12):2285–2308CrossRefGoogle Scholar
  53. Wijtten PJ, van der Meulen J, Verstegen MW (2011) Intestinal barrier function and absorption in pigs after weaning: a review. Br J Nutr 105(7):967–981CrossRefGoogle Scholar
  54. Wu G (2009) Amino acids: metabolism, functions, and nutrition. Amino Acids 37(1):1–17CrossRefGoogle Scholar
  55. Wu G (2010) Functional amino acids in growth, reproduction, and health. Adv Nutr 1(1):31–37CrossRefGoogle Scholar
  56. Wu G (2013) Functional amino acids in nutrition and health. Amino Acids 45(3):407–411CrossRefGoogle Scholar
  57. Wu G (2018) Principles of animal nutrition. CRC Press, Poca RatonGoogle Scholar
  58. Wu G, Meier SA, Knabe DA (1996) Dietary glutamine supplementation prevents jejunal atrophy in weaned pigs. J Nutr 126(10):2578–2584CrossRefGoogle Scholar
  59. Wu G, Knabe DA, Kim SW (2004) Arginine nutrition in neonatal pigs. J Nutr 134(10 Suppl):2783S–2790SCrossRefGoogle Scholar
  60. Wu G, Bazer FW, Dai Z, Li D, Wang J, Wu Z (2014) Amino acid nutrition in animals: protein synthesis and beyond. Annu Rev Anim Biosci 2:387–417CrossRefGoogle Scholar
  61. Wu T, Lv Y, Li X, Zhao D, Yi D, Wang L, Li P, Chen H, Hou Y, Gong J, Wu G (2018) Establishment of a recombinant Escherichia coli-induced piglet diarrhea model. Front Biosci (Landmark Ed) 23:1517–1534CrossRefGoogle Scholar
  62. Yang H, Xiong X, Wang X, Tan B, Li T, Yin Y (2016) Effects of weaning on intestinal upper villus epithelial cells of piglets. PLoS One 11(3):e0150216CrossRefGoogle Scholar
  63. Yi H, Jiang D, Zhang L, Xiong H, Han F, Wang Y (2016) Developmental expression of STATs, nuclear factor-kappaB and inflammatory genes in the jejunum of piglets during weaning. Int Immunopharmacol 36:199–204CrossRefGoogle Scholar
  64. Yi D, Hou Y, Xiao H, Wang L, Zhang Y, Chen H, Wu T, Ding B, Hu CA, Wu G (2017) N-Acetylcysteine improves intestinal function in lipopolysaccharides-challenged piglets through multiple signaling pathways. Amino Acids 49(12):1915–1929CrossRefGoogle Scholar
  65. Yi D, Li BC, Hou YQ, Wang L, Zhao D, Chen HB, Wu T, Zhou Y, Ding BY, Wu G (2018) Dietary supplementation with an amino acid blend enhances intestinal function in piglets. Amino Acids 50:1089–1100CrossRefGoogle Scholar
  66. Zhong Z, Wheeler MD, Li X, Froh M, Schemmer P, Yin M, Bunzendaul H, Bradford B, Lemasters JJ (2003) L-Glycine: a novel antiinflammatory, immunomodulatory, and cytoprotective agent. Curr Opin Clin Nutr Metab Care 6(2):229–240CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  • Xiaoxiao Fan
    • 1
    • 2
  • Shuai Li
    • 2
  • Zhenlong Wu
    • 1
    • 2
    Email author
  • Zhaolai Dai
    • 3
  • Ju Li
    • 2
  • Xiaolong Wang
    • 3
  • Guoyao Wu
    • 2
    • 4
  1. 1.Beijing Advanced Innovation Center for Food Nutrition and Human HealthChina Agricultural UniversityBeijingChina
  2. 2.Department of Animal Nutrition and Feed Science, State Key Laboratory of Animal NutritionChina Agricultural UniversityBeijingChina
  3. 3.Henan Yinfa Animal Husbandry CoXinzhengChina
  4. 4.Department of Animal ScienceTexas A&M UniversityCollege StationUSA

Personalised recommendations