Skip to main content

Advertisement

SpringerLink
Log in

The Generation of C-3α Epimer of 25-Hydroxyvitamin D and Its Biological Effects on Bone Mineral Density in Adult Rodents

  • Original Research
  • Published:
Calcified Tissue International Aims and scope Submit manuscript

Abstract

The source and function of C-3α epimer of 25(OH)D (C-3 epimer) is unknown. The objectives were to (1) establish if increasing doses of vitamin D (VD) results in a proportionate dose–response in C-3 epimer; and (2) determine the biological response of bone to C-3 epimer treatment. Sprague Dawley rats (12 weeks, n = 36 female, n = 36 male) were randomized to control AIN93-M diet (1 IU VD3/g diet) or an experimental diet for 8 weeks containing VD3 at 2 or 4 IU/g diet, C-3 epimer at 0.5 or 1 IU/g diet or 25(OH)D (0.5 IU/g diet). BW and food consumption were measured weekly. Blood was sampled at week 0, 4, and 8 for assessment of VD metabolites and bone metabolism biomarkers. DXA (week 0, 4, and 8) and in vivo micro CT (μCT) (week 0 and 8) were performed in vivo plus ex vivo μCT imaging and bone biomechanics. Dietary intake and anthropometry did not differ among diet groups. The dose–response of VD generated significantly elevated C-3 epimer only in females with concentrations of 4 IU VD diet group [mean 84.6 (62.5) nmol/L] exceeding control [mean 21.4 (18.5) nmol/L, p = 0.005]. Both sexes in the 25(OH)D group did not show significant increases in C-3 epimer, whereas 0.5 and 1 IU epimer groups exceeded 100 nmol/L of C-3 epimer by 8 weeks. These data suggest C-3 epimer is endogenously generated with higher intakes of VD. Endogenous and exogenous C-3 epimer accumulates in serum without impact upon bone health outcomes in a healthy young adult model over 8 weeks.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Bailey D, Veljkovic K, Yazdanpanah M, Adeli K (2013) Analytical measurement and clinical relevance of vitamin D(3) C3-epimer. Clin Biochem 46(3):190–196

    Article  CAS  PubMed  Google Scholar 

  2. Genazzani AR, Palumbo MA, de Micheroux AA, Artini PG, Criscuolo M, Ficarra G, Guo A-L, Benelli A, Bertolini A, Petraglia F (1995) Evidence for a role for the neurosteroid allopregnanolone in the modulation of reproductive function in female rats. Eur J Endocrinol 133(3):375–380

    Article  CAS  PubMed  Google Scholar 

  3. Reddy GS, Muralidharan KR, Okamura WH, Tserng KY, McLane JA (2001) Metabolism of 1alpha,25-dihydroxyvitamin D(3) and its C-3 epimer 1alpha,25-dihydroxy-3-epi-vitamin D(3) in neonatal human keratinocytes. Steroids 66(3–5):441–450

    Article  CAS  PubMed  Google Scholar 

  4. Kamao M, Hatakeyama S, Sakaki T, Sawada N, Inouye K, Kubodera N, Reddy GS, Okano T (2005) Measurement and characterization of C-3 epimerization activity toward vitamin D3. Arch Biochem Biophys 436(1):196–205

    Article  CAS  PubMed  Google Scholar 

  5. Fleet JC, Bradley J, Reddy GS, Ray R, Wood RJ (1996) 1 alpha,25-(OH)2-vitamin D3 analogs with minimal in vivo calcemic activity can stimulate significant transepithelial calcium transport and mRNA expression in vitro. Arch Biochem Biophys 329(2):228–234

    Article  CAS  PubMed  Google Scholar 

  6. Kamao M, Tatematsu S, Hatakeyama S, Sakaki T, Sawada N, Inouye K, Ozono K, Kubodera N, Reddy GS, Okano T (2004) C-3 epimerization of vitamin D3 metabolites and further metabolism of C-3 epimers: 25-hydroxyvitamin D3 is metabolized to 3-epi-25-hydroxyvitamin D3 and subsequently metabolized through C-1alpha or C-24 hydroxylation. J Biol Chem 279(16):15897–15907

    Article  CAS  PubMed  Google Scholar 

  7. Rehan VK, Torday JS, Peleg S, Gennaro L, Vouros P, Padbury J, Rao DS, Reddy GS (2002) 1Alpha,25-dihydroxy-3-epi-vitamin D3, a natural metabolite of 1alpha,25-dihydroxy vitamin D3: production and biological activity studies in pulmonary alveolar type II cells. Mol Genet Metab 76(1):46–56

    Article  CAS  PubMed  Google Scholar 

  8. Rhieu SY, Annalora AJ, Wang G, Flarakos CC, Gathungu RM, Vouros P, Sigueiro R, Mourino A, Schuster I, Palmore GT, Reddy GS (2013) Metabolic stability of 3-Epi-1alpha,25-dihydroxyvitamin D3 over 1 alpha 25-dihydroxyvitamin D3: metabolism and molecular docking studies using rat CYP24A1. J Cell Biochem 114(10):2293–2305

    Article  CAS  PubMed  Google Scholar 

  9. Gallo S, Comeau K, Vanstone C, Agellon S, Sharma A, Jones G, L’Abbe M, Khamessan A, Rodd C, Weiler H (2013) Effect of different dosages of oral vitamin D supplementation on vitamin D status in healthy, breastfed infants: a randomized trial. JAMA 309(17):1785–1792

    Article  CAS  PubMed  Google Scholar 

  10. Anderson PH, Sawyer RK, Moore AJ, May BK, O’Loughlin PD, Morris HA (2008) Vitamin D depletion induces RANKL-mediated osteoclastogenesis and bone loss in a rodent model. J Bone Miner Res 23(11):1789–1797

    Article  CAS  PubMed  Google Scholar 

  11. Lee AM, Anderson PH, Sawyer RK, Moore AJ, Forwood MR, Steck R, Morris HA, O’Loughlin PD (2010) Discordant effects of vitamin D deficiency in trabecular and cortical bone architecture and strength in growing rodents. J Steroid Biochem Mol Biol 121(1–2):284–287

    Article  CAS  PubMed  Google Scholar 

  12. Fleet JC, Gliniak C, Zhang Z, Xue Y, Smith KB, McCreedy R, Adedokun SA (2008) Serum metabolite profiles and target tissue gene expression define the effect of cholecalciferol intake on calcium metabolism in rats and mice. J Nutr 138(6):1114–1120

    PubMed Central  CAS  PubMed  Google Scholar 

  13. Pye KM, Wakefield AP, Aukema HM, House JD, Ogborn MR, Weiler HA (2009) A high mixed protein diet reduces body fat without altering the mechanical properties of bone in female rats. J Nutr 139(11):2099–2105

    Article  CAS  PubMed  Google Scholar 

  14. Reeves PG, Nielsen FH, Fahey GC Jr (1993) AIN-93 purified diets for laboratory rodents: final report of the American Institute of Nutrition ad hoc writing committee on the reformulation of the AIN-76A rodent diet. J Nutr 123(11):1939–1951

    CAS  PubMed  Google Scholar 

  15. Brouwers JE, Van Rietbergen B, Huiskes R (2007) No effects of in vivo micro-CT radiation on structural parameters and bone marrow cells in proximal tibia of wistar rats detected after eight weekly scans. J Orthop Res 25(10):1325–1332

    Article  PubMed  Google Scholar 

  16. Meganck JA, Kozloff KM, Thornton MM, Broski SM, Goldstein SA (2009) Beam hardening artifacts in micro-computed tomography scanning can be reduced by X-ray beam filtration and the resulting images can be used to accurately measure BMD. Bone 45(6):1104–1116

    Article  PubMed Central  PubMed  Google Scholar 

  17. Crenshaw TD, Peo ER, Lewis AJ, Moser BD (1981) Bone strength as a trait for assessing mineralization in swine: a critical review of techniques involved. J Anim Sci 53(3):827–835

    Google Scholar 

  18. Brown AJ, Ritter CS, Weiskopf AS, Vouros P, Sasso GJ, Uskokovic MR, Wang G, Reddy GS (2005) Isolation and identification of 1alpha-hydroxy-3-epi-vitamin D3, a potent suppressor of parathyroid hormone secretion. J Cell Biochem 96(3):569–578

    Article  CAS  PubMed  Google Scholar 

  19. Lensmeyer G, Poquette M, Wiebe D, Binkley N (2012) The C-3 epimer of 25-hydroxyvitamin D(3) is present in adult serum. J Clin Endocrinol Metabol 97(1):163–168

    Article  CAS  Google Scholar 

  20. Schleicher RL, Encisco SE, Chaudhary-Webb M, Paliakov E, McCoy LF, Pfeiffer CM (2011) Isotope dilution ultra performance liquid chromatography-tandem mass spectrometry method for simultaneous measurement of 25-hydroxyvitamin D2, 25-hydroxyvitamin D3 and 3-epi-25-hydroxyvitamin D3 in human serum. Clin Chim Acta 412(17–18):1594–1599

    Article  CAS  PubMed  Google Scholar 

  21. Yazdanpanah M, Bailey D, Walsh W, Wan B, Adeli K (2013) Analytical measurement of serum 25-OH-vitamin D(3), 25-OH-vitamin D(2) and their C3-epimers by LC-MS/MS in infant and pediatric specimens. Clin Biochem 46(13–14):1264–1271

    Article  CAS  PubMed  Google Scholar 

  22. Engelman CD, Bo R, Zuelsdorff M, Steltenpohl H, Kirby T, Nieto FJ (2014) Epidemiologic study of the C-3 epimer of 25-hydroxyvitamin D(3) in a population-based sample. Clin Nutr 33(3):421–425

    Article  CAS  PubMed  Google Scholar 

  23. Lensmeyer G, Poquette M, Wiebe D, Binkley N (2011) The C-3 epimer of 25-hydroxyvitamin D3 is present in adult serum. J Clin Endocrinol Metabol 97(1):163–168

    Article  Google Scholar 

  24. Witschi AK, Liesegang A, Gebert S, Weber GM, Wenk C (2011) Effect of source and quantity of dietary vitamin D in maternal and creep diets on bone metabolism and growth in piglets. J Anim Sci 89(6):1844–1852

    Article  CAS  PubMed  Google Scholar 

  25. Rhieu SY, Annalora AJ, Wang G, Flarakos CC, Gathungu RM, Vouros P, Sigueiro R, Mourino A, Schuster I, Palmore GT, Reddy GS (2013) Metabolic stability of 3-epi-1alpha,25-dihydroxyvitamin D3 over 1 alpha 25-dihydroxyvitamin D3: metabolism and molecular docking studies using rat CYP24A1. J Cell Biochem 114(10):2293–2305

    Article  CAS  PubMed  Google Scholar 

  26. Masuda S, Kamao M, Schroeder NJ, Makin HL, Jones G, Kremer R, Rhim J, Okano T (2000) Characterization of 3-epi-1alpha,25-dihydroxyvitamin D3 involved in 1alpha,25-dihydroxyvitamin D3 metabolic pathway in cultured cell lines. Biol Pharm Bull 23(2):133–139

    Article  CAS  PubMed  Google Scholar 

  27. Gallo S, Comeau K, Agellon S, Vanstone C, Sharma A, Jones G, L’Abbe M, Khamessan A, Weiler H, Rodd C (2014) Methodological issues in assessing plasma 25-hydroxyvitamin D concentration in newborn infants. Bone 61:186–190

    Article  CAS  PubMed  Google Scholar 

  28. Olfert ED, Cross BM, McWilliam AA (1993) Guide to the care and use of experimental animals, vol 2. Canadian Council on Animal Care, Ottawa

    Google Scholar 

Download references

Acknowledgments

This work was supported by a Collaborative Research and Development Grant from Dairy Farmers of Canada and the Natural Sciences and Engineering Research Council of Canada. Infrastructure support from the Canadian Foundation for Innovation and Canada Research Chairs programs awarded to HW is acknowledged. Vitamin D metabolites were measured by Warnex, Laval (Bioanalytical Services, Laval, QC). C-3 epimer and 25(OH)D was purchased from Isosciences (Isosciences, LCC. Pennsylvania, USA) for diet production by Harlan (Harlan Laboratories, Madison, WI). This study was reviewed and approved by the Macdonald Campus Facility Animal Care Committee and in accordance with the Canadian Council on Animal Care [28].

Conflict of Interest

Dr. Hope A. Weiler has received research Grants from a Collaborative Research and Development Grant from Dairy Farmers of Canada and the Natural Sciences and Engineering Research Council of Canada. Christina Bianchini, Paula Lavery, and Sherry Agellon declare they have no conflict of interest.

Human and Animal Rights and Informed Consent

This article does not contain any studies with human participants performed by any of the authors. All procedures performed in studies involving animals were reviewed and approved by the Macdonald Campus Facility Animal Care Committee and in accordance with the Canadian Council on Animal Care the ethical standards.

Author information

Authors and Affiliations

  1. Dietetics and Human Nutrition, McGill University, Macdonald-Stewart Building, Macdonald Campus, 21111 Lakeshore Road, Ste-Anne-de-Bellevue, Montréal, QC, H9X 3V9, Canada

    Christina Bianchini, Paula Lavery, Sherry Agellon & Hope A. Weiler

Authors
  1. Christina Bianchini
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Paula Lavery
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sherry Agellon
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hope A. Weiler
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hope A. Weiler.

Electronic Supplementary Material

Below is the link to the electronic supplementary material.

Supplementary material 1 (PDF 361 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bianchini, C., Lavery, P., Agellon, S. et al. The Generation of C-3α Epimer of 25-Hydroxyvitamin D and Its Biological Effects on Bone Mineral Density in Adult Rodents. Calcif Tissue Int 96, 453–464 (2015). https://doi.org/10.1007/s00223-015-9973-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00223-015-9973-9

Keywords

Search

Navigation