Maternal 25-hydroxycholecalciferol during lactation improves intestinal calcium absorption and bone properties in sow-suckling piglet pairs

  • Lianhua Zhang
  • Jiangxu Hu
  • Miao Li
  • Qinghui Shang
  • Sujie Liu
  • Xiangshu PiaoEmail author
Original Article


Lower maternal vitamin D status during lactation is a common health problem. The objectives of this study were to investigate the effects of maternal 25-hydroxycholecalciferol (25-OH-D3) supplementation during lactation on maternal and neonatal bone health in a sow model. 32 Large White × Landrace sows were assigned randomly to one of two diets supplemented with 2000 IU/kg vitamin D3 (ND) or 50 μg/kg 25-OH-D3 (25-D). The experiment began on day 107 of gestation and continued until weaning on day 21 of lactation. Maternal 25-OH-D3 supplementation significantly decreased milk n-6:n-3 PUFA ratio, which supported bone formation of piglets. Supplementation with 25-OH-D3 altered bone turnover rate of sows and piglets, as evidenced by higher bone-specific alkaline phosphatase (BALP) concentration in serum. 25-D sows had significantly higher bone density and mechanical properties of tibias and femurs than ND sows. Calcium (Ca) absorption rate was higher in 25-D sows than ND sows, which was caused partially by the increased mRNA expressions of renal 1α-hydroxylase (CYP27B1) and duodenal vitamin D receptor (VDR), transient receptor potential vanilloid 6 (TRPV6), and calcium-binding protein D9k (CaBP-D9k). Maternal 25-OH-D3 supplementation increased tibial and femoral Ca content by up-regulating Ca-related gene expression in kidney (CYP27B1), ileum (VDR and claudin-2), and colon (VDR and CaBP-D9k), thus, activating 1,25-dihydroxyvitamin D3 [1,25-(OH)2-D3]-dependent Ca transport in piglets. In conclusion, improved milk fatty acids and higher mRNA expressions of calcitropic genes triggered by maternal 25-OH-D3 supplementation would be the potential mechanism underlying the positive effects of 25-OH-D3 on maternal and neonatal bone health.


Maternal 25-hydroxycholecalciferol Milk fatty acids Calcium absorption Bone Piglets 



This research was financially supported by the National Natural Science Foundation of China (No. 31772612). We gratefully acknowledge Haineng Bioengineering Co., Ltd. (Rizhao, China) who provides us with experimental product and scientific funding used in this study.

Author contributions

The authors’ contributions are as follows: XP and LZ designed research; LZ carried out the experiment design and wrote the draft of manuscript; LZ, JH, and ML were responsible for the data analysis; LZ, QS, and SL reviewed and revised the manuscript; XP had primary responsibility for final content. All authors read and approved the final manuscript.

Compliance with ethical standards

Conflict of interest

All authors have no conflicts of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

Supplementary material

774_2019_1020_MOESM1_ESM.docx (21 kb)
Supplementary material 1 (DOCX 21 kb)


  1. 1.
    Park JW, Lee JH, Kim SW, Han JS, Kang KS, Kim SJ, Park TS (2018) Muscle differentiation induced up-regulation of calcium-related gene expression in quail myoblasts. Asian-Australas J Anim Sci 31:1507–1515CrossRefGoogle Scholar
  2. 2.
    Brun LR, Lombarte M, Roma S, Perez F, Millán JL, Rigalli A (2018) Increased calcium uptake and improved trabecular bone properties in intestinal alkaline phosphatase knockout mice. J Bone Miner Metab 36:661–667CrossRefGoogle Scholar
  3. 3.
    Kovacs CS (2016) Maternal mineral and bone metabolism during pregnancy, lactation, and post-weaning recovery. Physiol Rev 96:449–547CrossRefGoogle Scholar
  4. 4.
    Halloran BP, Barthell E, DeLuca HF (1979) Vitamin D metabolism during pregnancy and lactation in the rat. Proc Natl Acad Sci 76:5549–5553CrossRefGoogle Scholar
  5. 5.
    Cross NA, Hillman LS, Allen SH, Krause GF, Vieira NE (1995) Calcium homeostasis and bone metabolism during pregnancy, lactation, and postweaning: a longitudinal study. Am J Clin Nutr 61:514–523CrossRefGoogle Scholar
  6. 6.
    Costa ML, Krupa FG, Rehder PM, Sousa MH, Costa-Paiva L, Cecatti JG (2012) Forearm bone mineral density changes during postpartum and the effects of breastfeeding, amenorrhea, body mass index and contraceptive use. Osteoporos Int 23:1691–1698CrossRefGoogle Scholar
  7. 7.
    Kalkwarf HJ, Specker BL, Bianchi DC, Ranz J, Ho M (1997) The effect of calcium supplementation on bone density during lactation and after weaning. N Engl J Med 21:523–528CrossRefGoogle Scholar
  8. 8.
    Choi SW, Kweon SS, Choi JS, Rhee JA, Lee YH, Nam HS, Jeong SK, Park KS, Ryu SY, Song HR, Shin MH (2016) The association between vitamin D and parathyroid hormone and bone mineral density: the Dong-gu Study. J Bone Miner Metab 34:555–563CrossRefGoogle Scholar
  9. 9.
    Jakobsen J, Maribo H, Bysted A, Sommer HM, Hels O (2007) 25-hydroxyvitamin D3 affects vitamin D status similar to vitamin D3 in pigs–but the meat produced has a lower content of vitamin D. Br J Nutr 98:908–913CrossRefGoogle Scholar
  10. 10.
    Kimball S, Gel-H Fuleihan, Vieth R (2008) Vitamin D: a growing perspective. Crit Rev Clin Lab Sci 45:339–414CrossRefGoogle Scholar
  11. 11.
    Pilz S, Zittermann A, Obeid R, Hahn A, Pludowski P, Trummer C, Lerchbaum E, Pérez-López FR, Karras SN, März W (2018) The role of vitamin D in fertility and during pregnancy and lactation: a review of clinical data. Int J Environ Res Public Health 15:2241CrossRefGoogle Scholar
  12. 12.
    Zinser GM, Welsh J (2004) Accelerated mammary gland development during pregnancy and delayed postlactational involution in vitamin D3 receptor null mice. Mol Endocrinol 18:2208–2223CrossRefGoogle Scholar
  13. 13.
    Pludowski P, Holick MF, Pilz S, Wagner CL, Hollis BW, Grant WB, Shoenfeld Y, Lerchbaum E, Llewellyn DJ, Kienreich K, Soni M (2013) Vitamin D effects on musculoskeletal health, immunity, autoimmunity, cardiovascular disease, cancer, fertility, pregnancy, dementia and mortality-a review of recent evidence. Autoimmun Rev 12:976–989CrossRefGoogle Scholar
  14. 14.
    Cashman KD, Seamans KM, Lucey AJ, Stöcklin E, Weber P, Kiely M, Hill TR (2012) Relative effectiveness of oral 25-hydroxyvitamin D3 and vitamin D3 in raising wintertime serum 25-hydroxyvitamin D in older adults. Am J Clin Nutr 95:1350–1356CrossRefGoogle Scholar
  15. 15.
    National Research Council (2012) Nutrient requirements of swine. National Academy Press, Washington, DCGoogle Scholar
  16. 16.
    Xu Y, Zeng Z, Xu X, Tian Q, Ma X, Long S, Piao M, Cheng Z, Piao X (2017) Effects of the standardized ileal digestible valine:lysine ratio on performance, milk composition and plasma indices of lactating sows. Anim Sci J 88:1082–1092CrossRefGoogle Scholar
  17. 17.
    Gomes FP, Shaw PN, Whitfield K, Hewavitharana AK (2015) Simultaneous quantitative analysis of eight vitamin D analogues in milk using liquid chromatography-tandem mass spectrometry. Anal Chim Acta 891:211–220CrossRefGoogle Scholar
  18. 18.
    Jang YD, Lindemann MD, Agudelo-Trujillo JH, Escobar CS, Kerr BJ, Inocencio N, Cromwell GL (2014) Comparison of direct and indirect estimates of apparent total tract digestibility in swine with effort to reduce variation by pooling of multiple day fecal samples. J Anim Sci 92:4566–4576CrossRefGoogle Scholar
  19. 19.
    Association of Official Analytical Chemists (2006) Official methods of analysis. AOAC Int, Gaithersgurg, VAGoogle Scholar
  20. 20.
    Keenan MJ, Hegsted M, Jones KL, Delany JP, Kime JC, Melancon LE, Tulley RT, Hong KD (1997) Comparison of bone density measurement techniques: DXA and Archimedes’ principle. J Bone Miner Res 12:1903–1907CrossRefGoogle Scholar
  21. 21.
    Flohr JR, Tokach MD, Dritz SS, DeRouchey JM, Goodband RD, Nelssen JL, Bergstrom JR (2014) An evaluation of the effects of added vitamin D3 in maternal diets on sow and pig performance. J Anim Sci 92:594–603CrossRefGoogle Scholar
  22. 22.
    Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408CrossRefGoogle Scholar
  23. 23.
    Swindle MM, Makin A, Herron AJ, Clubb FJ Jr, Frazier KS (2012) Swine as models in biomedical research and toxicology testing. Vet Pathol 49:344–356CrossRefGoogle Scholar
  24. 24.
    Lauridsen C, Halekoh U, Larsen T, Jensen SK (2010) Reproductive performance and bone status markers of gilts and lactating sows supplemented with two different forms of vitamin D. J Anim Sci 88:202–213CrossRefGoogle Scholar
  25. 25.
    Goff JP, Horst RL, Littledike ET (1984) Effect of sow vitamin D status at parturition on the vitamin D status of neonatal piglets. J Nutr 114:163–169CrossRefGoogle Scholar
  26. 26.
    Hollis BW, Wagner CL (2004) Assessment of dietary vitamin D requirements during pregnancy and lactation. Am J Clin Nutr 79:717–726Google Scholar
  27. 27.
    Specker BL, Tsang RC, Hollis BW (1985) Effect of race and diet on human-milk vitamin D and 25-hydroxyvitamin D. Am J Dis Child 139:1134–1137Google Scholar
  28. 28.
    Weber GM, Witschi AK, Wenk C, Martens H (2014) Effects of dietary 25-hydroxycholecalciferol and cholecalciferol on blood vitamin D and mineral status, bone turnover, milk composition, and reproductive performance of sows. J Anim Sci 92:899–909CrossRefGoogle Scholar
  29. 29.
    Li Y, Seifert MF, Ney DM, Grahn M, Grant AL, Allen KG, Watkins BA (1999) Dietary conjugated linoleic acids alter serum IGF-I and IGF binding protein concentrations and reduce bone formation in rats fed (n-6) or (n-3) fatty acids. J Bone Miner Res 14:1153–1162CrossRefGoogle Scholar
  30. 30.
    Korotkova M, Ohlsson C, Hanson LA, Strandvik B (2004) Dietary n-6:n-3 fatty acid ratio in the perinatal period affects bone parameters in adult female rats. Br J Nutr 92:643–648CrossRefGoogle Scholar
  31. 31.
    Watkins BA, Li Y, Allen KG, Hoffmann WE, Seifert MF (2000) Dietary ratio of (n-6)/(n-3) polyunsaturated fatty acids alters the fatty acid composition of bone compartments and biomarkers of bone formation in rats. J Nutr 130:2274–2284CrossRefGoogle Scholar
  32. 32.
    Zinser GM, Welsh J (2004) Accelerated mammary gland development during pregnancy and delayed postlactational involution in vitamin D3 receptor null mice. Mol Endocrinol 18:2208–2223CrossRefGoogle Scholar
  33. 33.
    Allen JC (1984) Effect of vitamin D deficiency on mouse mammary gland and milk. J Nutr 114:42–49CrossRefGoogle Scholar
  34. 34.
    Kang EJ, Lee JE, An SM, Lee JH, Kwon HS, Kim BC, Kim SJ, Kim JM, Hwang DY, Jung YJ, Yang SY, Kim SC, An BS (2015) The effects of vitamin D3 on lipogenesis in the liver and adipose tissue of pregnant rats. Int J Mol Med 36:1151–1158CrossRefGoogle Scholar
  35. 35.
    Ji L, Gupta M, Feldman BJ (2016) Vitamin D regulates fatty acid composition in subcutaneous adipose tissue through Elovl3. Endocrinology 157:91–97CrossRefGoogle Scholar
  36. 36.
    Ram VS, Parthiban Sudhakar U, Mithradas N, Prabhakar R (2015) Bone biomarkers in periodontal disease: a review article. J Clin Diagn Res 9:ZE07–10Google Scholar
  37. 37.
    Harada K, Itoh H, Kawazoe Y, Miyazaki S, Doi K, Kubo T, Akagawa Y, Shiba T (2013) Polyphosphate-mediated inhibition of tartrate-resistant acid phosphatase and suppression of bone resorption of osteoclasts. PLoS One 8:e78612CrossRefGoogle Scholar
  38. 38.
    Bogazzi F, Rossi G, Lombardi M, Tomisti L, Sardella C, Manetti L, Curzio O, Marcocci C, Grasso L, Gasperi M, Martino E (2011) Vitamin D status may contribute to serum insulin-like growth factor I concentrations in healthy subjects. J Endocrinol Invest 34:e200–e203CrossRefGoogle Scholar
  39. 39.
    Ohlsson C, Bengtsson BA, Isaksson OG, Andreassen TT, Slootweg MC (1998) Growth hormone and bone. Endocr Rev 19:55–79Google Scholar
  40. 40.
    Haussler MR, Whitfield GK, Kaneko I, Haussler CA, Hsieh D, Hsieh JC, Jurutka PW (2013) Molecular mechanisms of vitamin D action. Calcif Tissue Int 92:77–98CrossRefGoogle Scholar
  41. 41.
    Christakos S, Dhawan P, Porta A, Mady LJ, Seth T (2011) Vitamin D and intestinal calcium absorption. Mol Cell Endocrinol 347:25–29CrossRefGoogle Scholar
  42. 42.
    Bronner F, Pansu D (1999) Nutritional aspects of calcium absorption. J Nutr 129:9–12CrossRefGoogle Scholar
  43. 43.
    Mahan DC, Vallet JL (1997) Vitamin and mineral transfer during fetal development and the early postnatal period in pigs. J Anim Sci 75:2731–2738CrossRefGoogle Scholar
  44. 44.
    Alexander RT, Rievaj J, Dimke H (2014) Paracellular calcium transport across renal and intestinal epithelia. Biochem Cell Biol 92:467–480CrossRefGoogle Scholar
  45. 45.
    Pointillart A, Coxam V, Sève B, Colin C, Lacroix CH, Guéguen L (2000) Availability of calcium from skim milk, calcium sulfate and calcium carbonate for bone mineralization in pigs. Reprod Nutr Dev 40:49–61CrossRefGoogle Scholar

Copyright information

© Springer Japan KK, part of Springer Nature 2019

Authors and Affiliations

  1. 1.State Key Laboratory of Animal Nutrition, College of Animal Science and TechnologyChina Agricultural UniversityBeijingChina

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