Korean J Nutr.  2009 Jun;42(4):309-317. 10.4163/kjn.2009.42.4.309.

Effects of Arginine Supplementation on Bone Mineral Density and Bone Markers in OVX Rats

• Choi MJ

• KMID: 2265638
• DOI: http://doi.org/10.4163/kjn.2009.42.4.309

Abstract

As far as we know, there were no studies of the effect of L-arginine on bone metabolism in post-menopausal women or ovariectomized rats. The primary objective of the current study was to determine whether arginine supplementation was associated with alterations in femoral and spinal bone mineral density (BMD) and bone markers in ovariectomized (Ovx) rats. Forty female Sprague-Dawley rats were divided into two groups, Ovx and sham groups, which were each randomly divided into two subgroups that were fed control and arginine supplemented diet. All rats were fed on experimental diet and deionized water ad libitum for 9 weeks. Bone formation was measured by serum osteocalcin and alkaline phosphatase (ALP) concentrations. Bone resorption was measured by deoxypyridinoline (DPD) crosslinks immunoassay and corrected for creatinine. Serum osteocalcin, growth hormone, insulin-like growth factor-1 (IGF-1), parathyroid hormone (PTH) and calcitonin were analyzed using radioimmunoassay kits. Bone mineral density (BMD) and bone mineral content (BMC) were measured using PIXImus (GE Lunar Co, Wisconsin, USA) in spine and femur. The serum and urine concentrations of Ca and P were determined. The plasma was analyzed for arginine. Diet did not affect weight gain, mean food intake, and plasma arginine concentration. Urinary Ca excretion was decreased by arginine supplementation in Ovx rats, but statistically not significant. The Ovx rats fed arginine-supplemented diet were not significantly different in ALP, osteocalcin, crosslinks value, PTH, calcitonin and IGF-1 compared to those fed control diet. The arginine-supplemented group had significantly higher serum Ca and growth hormone than control group. Spine and femur BMD were significantly increased by arginine supplementation on 5th and 9th weeks after feeding. Our findings indicate that dietary L-arginine supplementation decreased bone mineral density loss in Ovx rats. Therefore, dietary arginine supplementation may represent a potentially useful strategy for the management of osteoporosis.

Keyword

arginine; BMD; GH; IGF-1; PTH; bone markers; ovariectomized rat

MeSH Terms

Alkaline Phosphatase
Amino Acids
Animals
Arginine
Bone Density
Bone Resorption
Calcitonin
Creatinine
Diet
Eating
Female
Femur
Growth Hormone
Humans
Immunoassay
Insulin-Like Growth Factor I
Osteocalcin
Osteogenesis
Osteoporosis
Parathyroid Hormone
Plasma
Radioimmunoassay
Rats
Rats, Sprague-Dawley
Salicylamides
Spine
Water
Weight Gain
Wisconsin
Alkaline Phosphatase
Amino Acids
Arginine
Calcitonin
Creatinine
Growth Hormone
Insulin-Like Growth Factor I
Osteocalcin
Parathyroid Hormone
Salicylamides
Water

Reference

1. Greendale GA, FitzGerald G, Huang MH, Sternfeld B, Gold E, Seeman T, Sherman S, Sowers M. Dietary soy isoflavones and bone mineral density: results from the study of women's health across the nation. Am J Epidemiol. 2002. 155(8):746–754.

2. Ho SC, Chan SG, Yi Q, Wong E, Leung PC. Soy intake and the maintenance of peak bone mass in Hong Kong Chinese women. J Bone Miner Res. 2001. 16:1363–1369.

3. Setchell KD, Lydeking-Olsen E. Dietary phytoestrogens and their effect on bone: evidence from in vitro and in vivo, human observational, and dietary intervention studies. Am J Clin Nutr. 2003. 78:593S–609S.

4. Atkinson C, Compston JE, Day NE, Dowsett M, Bingham SA. The effects of phytoestrogen isoflavones on bone density in women: a double-blind, randomized, placebo-controlled trial. Am J Clin Nutr. 2004. 79:326–333.

5. Spence LA, Lipscomb ER, Cadogan J, Martin B, Wastney ME, Peacock M, Weaver CM. The effect of soy protein and soy isoflavones on calcium metabolism in postmenopausal women: a randomized crossover study. Am J Clin Nutr. 2005. 81:916–922.

6. Andreassen TT, Jorgensen PH, Flyvbjerg A, Orskov H, Oxlund H. Growth hormone stimulates bone formation and strength of cortical bone in aged rats. J Bone Miner Res. 1995. 10:1057–1067.

7. Monson JP, Arake WM, Carroll PV, Weaver JU, Rodriguez-Arnao J, Savage MO. Influence of growth hormone on accretion of bone mass. Horm Res. 2000. 58:Suppl 1. 52s–56s.

8. Sugimoto T, Kaji H, Nakaoka D, Yamauchi M, Tano S, Sugishita T, Baylink DJ, Mohan S, Chihara K. Effect of low-dose of recombinant human growth hormone on bone metabolism in elderly women with osteoporosis. Eur J Endocrinol. 2002. 147(3):339–348.

9. Choi MJ. Effects of arginine supplementation on bone mineral density in growing female rats. Korean J Nutr. 2007. 40(3):235–241.

10. Reeves PG, Nielsen FH, Fahey GC. AIN-93 purified diets for laboratory rodents. J Nutr. 1993. 123:1939–1951.

11. Xing S, Cekan SZ, Dicafalusy U. Validation of radioimunoassay for estadiol-17 isotope dilution-mass spectrometry and a test of radiochemical purity. Clin Chem Acta. 1983. 135:189–201.

12. Guarnero P, Grimaux M, Seguin P, Delmas P. Characterization of immunoreactive forms of human osteocalcin generated in vivo and in vitro. J Bone Min Res. 1994. 9:692–698.

13. Nanda N, Joshi H, Subbarao SK, Sharma VP. Two-site immunoradiometricassay (IRMA): detection, efficiency, and procedural modifications. J Am Mosq Control Assoc. 1994. 10:225–227.

14. Guarnero P, Grimaux M, Seguin P, Delmas P. Characterization of immunoreactive forms of human osteocalcin generated in vivo and in vitro. J Bone Min Res. 1994. 9:692–698.

15. Luiking Yvette C., Deutz Nicolaas E.P. Biomarkers of arginine and lysine excess. J Nutr. 2007. 137:1662S–1666S.

16. Daly JM, Reynolds J, Thom A, Kinsley L, Dietrick-Gallagher M, Shou J, Ruggieri B. Immune and metabolic effects of arginine in the surgical patient. Ann Surg. 1988. 208:512–523.

17. Chyun JH, Griminger P. Improvement of nitrogen retention by arginine and glycine supplementation and its relation to collagen synthesis in traumatized mature and aged rats. J Nutr. 1984. 114:1697–1704.

18. Kohli R, Meininger CJ, Haynes TE, Yan W, Self JT, Wu G. Dietary L-arginine supplementation enhances endothelial nitric oxide synthesis in streptozotocin-induced diabetic rats. J Nutr. 2004. 134(3):600–608.

19. Mituka BM, Rawnsley HN. Clinical biochemical and hematological reference value in normal experimental animals and normal humans. 1987. 2nd edition. New York: Masson;160.

20. Choi MJ. Effects of soy protein on bone mineral content and bone mineral density in growing male rats. Korean J Nutr. 2002. 35(4):409–413.

21. Choi MJ, Jo HJ. Effects of soy protein and isoflavones on bone mineral density in growing female rats. Korean J Nutr. 2003. 36(4):359–367.

22. Choi MJ, Jung SH. The effect of dietary source and sulfur amino acid content on bone metabolism in growing rats. Korean J Nutr. 2004. 37(2):100–107.

23. Choi MJ. Effects of soy and isoflavones on bone markers and hormones in growing male rats. Korean J Nutr. 2003. 36(5):452–458.

24. Choi MJ, Kim JH, Chang KJ. The effect of dietary taurine supplementation on plasma and liver lipid concentrations and free amino acid concentrations in rats fed a high-cholesterol diet. Adv Exp Med Biol. 2006. 583:235–242.

25. Lee YM, Choi MJ, Chang KJ. The effect of dietary taurine supplementation on plasma and urinary free amino acid concentrations in diabetic rats. Adv Exp Med Biol. 2003. 526:75–82.

26. Choi MJ. Effects of soy and isoflavones on bone markers and hormones in growing male rats. Korean J Nutr. 2003. 36(5):452–458.

27. Kalu DN, Masoro EJ, Yu BP, Hardin RR, Hollis BW. Modulation of age related hyperparathyroidism and senile bone loss in fischer rats by soy protein and food restriction. Endocrinology. 1998. 122:1847–1854.

28. Choi MJ, Cho HJ. Effects of soy and isoflavones on bone metabolism in growing female rats. Korean J Nutr. 2003. 36(6):549–558.

29. Bravenboer N, Hoizmann PJ, Matten JC, Stuurman LM, Roos JC, Lips P. Effect of long-term growth hormone treatment on bone mass and bone metabolism in growth hormone-deficient men. J Bone Miner Res. 2005. 20(10):1778–1784.

30. Dean HJ, Kellett JG, Bala RM. The effect of growth hormone treatment on somatomedin levels in growth hormone deficient children. J Clin Endocrinol Metab. 1982. 55:1167–1173.

31. Lely AJ. Growth hormone and aging. GH & IGF Res. 1999. 9:117–119.

32. Lewinson D, Shenzer P, Hochberg Z. Growth hormone involvement in the regulation of tartrate resistant acid phosphatase positive cells that are active in cartilage and bone resorption. Calcif Tissue Int. 1993. 52:216–221.

33. Choi MJ. Effects of arginine supplementation on bone markers and hormones in growing female rats. Korean J Nutr. 2007. 40(4):320–326.

34. Barret-Connor E, Goodman-Gruen D. Gender differences in insulin-like growth factor and bone mineral density association in old age: the Rancho Bernardo Study. J Bone Miner Res. 1998. 13(8):1343–1349.

35. Sugimoto T, Nishiyama K, Kuribayashi F, Chihara K. Serum levels of insulin-like growth factor (IGF) I, IGF binding protein (IGFBP)-2 and IGFBP-3 in osteoporotic patients with and without spinal fractures. J Bone Miner Res. 1997. 12:1272–1279.

36. Krassas GE, Papadopoulou PH, Koliakos G. Growth hormone, insulin growth factor-I, and IGF binding protein-3 axis relationship with bone mineral density among healthy men. Arch Androl. 2003. 49:191–199.