Horm Metab Res 2016; 48(07): 446-451
DOI: 10.1055/s-0042-104060
Endocrine Care

The Effects of Vitamin D-K-Calcium Co-Supplementation on Endocrine, Inflammation, and Oxidative Stress Biomarkers in Vitamin D-Deficient Women with Polycystic Ovary Syndrome: A Randomized, Double-Blind, Placebo-Controlled Trial

M. Razavi
1   Department of Gynecology and Obstetrics, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran
M. Jamilian
2   Endocrinology and Metabolism Research Center, Arak University of Medical Sciences, Arak, Iran
3   Department of Gynecology and Obstetrics, School of Medicine, Arak University of Medical Sciences, Arak, Iran
M. Karamali
4   Department of Gynecology and Obstetrics, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
F. Bahmani
5   Research Center for Biochemistry and Nutrition in Metabolic Diseases, Kashan University of Medical Sciences, Kashan, Iran
E. Aghadavod
5   Research Center for Biochemistry and Nutrition in Metabolic Diseases, Kashan University of Medical Sciences, Kashan, Iran
Z. Asemi
5   Research Center for Biochemistry and Nutrition in Metabolic Diseases, Kashan University of Medical Sciences, Kashan, Iran
› Author Affiliations


The current study was conducted to assess the effects of vitamin D-K-calcium co-supplementation on endocrine, inflammation, and oxidative stress biomarkers in vitamin D-deficient women with polycystic ovary syndrome (PCOS). This randomized double-blind, placebo-controlled trial was performed on 60 vitamin D-deficient women diagnosed with PCOS aged 18–40 years old. Participants were randomly allocated into 2 groups to intake either 200 IU vitamin D, 90 μg vitamin K plus, 500 mg calcium supplements (n=30), or placebo (n=30) twice a day for 8 weeks. Endocrine, inflammation, and oxidative stress biomarkers were quantified at the beginning and the end of the study. After 8 weeks of intervention, compared with the placebo, vitamin D-K-calcium co-supplementation resulted in a significant reduction in serum-free testosterone (− 2.1±1.6 vs.+0.1±1.0 pg/ml, p<0.001) and dehydroepiandrosterone sulfate (DHEAS) levels (− 0.8±1.0 vs.−0.1±0.5 μg/ml, p=0.006). In addition, a significant increase in plasma total antioxidant capacity (TAC) (+ 75.7±126.1 vs.−80.4±242.8 mmol/l, p=0.005) and a significant difference in plasma malondialdehyde (MDA) concentrations (+ 0.03±0.6 vs.+1.4±2.4 μmol/l, p=0.005) was observed following the supplementation with vitamin D-K-calcium compared with the placebo. A trend toward a greater decrease in luteinizing hormone was observed in vitamin D-K-calcium co-supplement group compared to placebo group (− 7.0 vs.−1.2 IU/l, p=0.09). We did not find any significant effect of vitamin D-K-calcium co-supplementation on prolactin, follicle-stimulating hormone, 17-OH progesterone, inflammatory markers, and glutathione levels. Overall, vitamin D-K-calcium co-supplementation for 8 weeks among vitamin D-deficient women with PCOS had beneficial effects on serum DHEAS, free testosterone, plasma TAC, and MDA levels.

Publication History

Received: 26 November 2015

Accepted: 24 February 2016

Article published online:
06 April 2016

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

  • References

  • 1 Trivax B, Azziz R. Diagnosis of polycystic ovary syndrome. Clin Obstet Gynecol 2007; 50: 168-177
  • 2 Rosenfield RL. The Polycystic Ovary Morphology-Polycystic Ovary Syndrome Spectrum. J Pediatr Adolesc Gynecol 2015; 28: 412-419
  • 3 Azziz R, Sanchez LA, Knochenhauer ES, Moran C, Lazenby J, Stephens KC, Taylor K, Boots LR. Androgen excess in women: experience with over 1000 consecutive patients. J Clin Endocrinol Metab 2004; 89: 453-462
  • 4 Escobar-Morreale HF, Luque-Ramirez M, Gonzalez F. Circulating inflammatory markers in polycystic ovary syndrome: a systematic review and metaanalysis. Fertil Steril 2011; 95: 1048-1058 e1–2
  • 5 Showell MG, Brown J, Clarke J, Hart RJ. Antioxidants for female subfertility. Cochrane Database Syst Rev 2013; 8: CD007807
  • 6 Zheng SH, Li XL. Visceral adiposity index as a predictor of clinical severity and therapeutic outcome of PCOS. Gynecol Endocrinol 2015; 1-7
  • 7 Hyderali BN, Mala K. Oxidative stress and cardiovascular complications in polycystic ovarian syndrome. Eur J Obstet Gynecol Reprod Biol 2015; 191: 15-22
  • 8 Chakraborty P, Ghosh S, Goswami SK, Kabir SN, Chakravarty B, Jana K. Altered trace mineral milieu might play an aetiological role in the pathogenesis of polycystic ovary syndrome. Biol Trace Elem Res 2013; 152: 9-15
  • 9 Kim JJ, Choi YM, Chae SJ, Hwang KR, Yoon SH, Kim MJ, Kim SM, Ku SY, Kim SH, Kim JG. Vitamin D deficiency in women with polycystic ovary syndrome. Clin Exp Reprod Med 2014; 41: 80-85
  • 10 Pal L, Berry A, Coraluzzi L, Kustan E, Danton C, Shaw J, Taylor H. Therapeutic implications of vitamin D and calcium in overweight women with polycystic ovary syndrome. Gynecol Endocrinol 2012; 28: 965-968
  • 11 Foroozanfard F, Jamilian M, Bahmani F, Talaee R, Talaee N, Hashemi T, Nasri K, Asemi Z, Esmaillzadeh A. Calcium plus vitamin D supplementation influences biomarkers of inflammation and oxidative stress in overweight and vitamin D-deficient women with polycystic ovary syndrome: a randomized double-blind placebo-controlled clinical trial. Clin Endocrinol (Oxf) 2015; 83: 888-894
  • 12 Mazzanti L, Battino M, Nanetti L, Raffaelli F, Alidori A, Sforza G, Carle F, Quagliarini V, Cester N, Vignini A. Effect of 1-year dietary supplementation with vitaminized olive oil on markers of bone turnover and oxidative stress in healthy post-menopausal women. Endocrine 2015; 50: 326-334
  • 13 Tehrani HG, Mostajeran F, Shahsavari S. The effect of calcium and vitamin D supplementation on menstrual cycle, body mass index and hyperandrogenism state of women with poly cystic ovarian syndrome. J Res Med Sci 2014; 19: 875-880
  • 14 Firouzabadi R, Aflatoonian A, Modarresi S, Sekhavat L, MohammadTaheri S. Therapeutic effects of calcium & vitamin D supplementation in women with PCOS. Complement Ther Clin Pract 2012; 18: 85-88
  • 15 Lerchbaum E, Rabe T. Vitamin D and female fertility. Curr Opin Obstet Gynecol 2014; 26: 145-150
  • 16 Ullah G, Jung P, Machaca K. Modeling Ca2+signaling differentiation during oocyte maturation. Cell Calcium 2007; 42: 556-564
  • 17 Juanola-Falgarona M, Salas-Salvado J, Estruch R, Portillo MP, Casas R, Miranda J, Martinez-Gonzalez MA, Bullo M. Association between dietary phylloquinone intake and peripheral metabolic risk markers related to insulin resistance and diabetes in elderly subjects at high cardiovascular risk. Cardiovasc Diabetol 2013; 12: 7
  • 18 Rotterdam ESHRE/ASRM-Sponsored PCOS Consensus Workshop Group. Revised 2003 consensus on diagnostic criteria and long-term health risks related to polycystic ovary syndrome. Fertil Steril 2004; 81: 19-25
  • 19 Huang A, Brennan K, Azziz R. Prevalence of hyperandrogenemia in the polycystic ovary syndrome diagnosed by the National Institutes of Health 1990 criteria. Fertil Steril 2010; 93: 1938-1941
  • 20 Irani M, Seifer DB, Grazi RV, Julka N, Bhatt D, Kalgi B, Irani S, Tal O, Lambert-Messerlian G, Tal R. Vitamin D Supplementation Decreases TGF-beta1 Bioavailability in PCOS: A Randomized Placebo-Controlled Trial. J Clin Endocrinol Metab 2015; 100: 4307-4314
  • 21 Jamil AS, Alalaf SK, Al-Tawil NG, Al-Shawaf T. A case-control observational study of insulin resistance and metabolic syndrome among the four phenotypes of polycystic ovary syndrome based on Rotterdam criteria. Reprod Health 2015; 12: 7
  • 22 Jamilian M, Foroozanfard F, Bahmani F, Talaee R, Monavari M, Asemi Z. Effects of Zinc Supplementation on Endocrine Outcomes in Women with Polycystic Ovary Syndrome: a Randomized, Double-Blind, Placebo-Controlled Trial. Biol Trace Elem Res. 2015 Aug 28 [Epub ahead of print]
  • 23 Tatsch E, Bochi GV, Pereira Rda S, Kober H, Agertt VA, de Campos MM, Gomes P, Duarte MM, Moresco RN. A simple and inexpensive automated technique for measurement of serum nitrite/nitrate. Clin Biochem 2011; 44: 348-350
  • 24 Benzie IF, Strain JJ. The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: the FRAP assay. Anal Biochem 1996; 239: 70-76
  • 25 Beutler E, Gelbart T. Plasma glutathione in health and in patients with malignant disease. J Lab Clin Med 1985; 105: 581-584
  • 26 Janero DR. Malondialdehyde and thiobarbituric acid-reactivity as diagnostic indices of lipid peroxidation and peroxidative tissue injury. Free Radic Biol Med 1990; 9: 515-540
  • 27 Razavi M, Jamilian M, Kashan ZF, Heidar Z, Mohseni M, Ghandi Y, Bagherian T, Asemi Z. Selenium Supplementation and the Effects on Reproductive Outcomes, Biomarkers of Inflammation, and Oxidative Stress in Women with Polycystic Ovary Syndrome. Horm Metab Res. 2015 Aug. 12 [Epub ahead of print]
  • 28 Asemi Z, Esmaillzadeh A. DASH diet, insulin resistance, and serum hs-CRP in polycystic ovary syndrome: a randomized controlled clinical trial. Horm Metab Res 2015; 47: 232-238
  • 29 Shah S, Wilson DM, Bachrach LK. Large Doses of Vitamin D Fail to Increase 25-Hydroxyvitamin D Levels or to Alter Cardiovascular Risk Factors in Obese Adolescents: A Pilot Study. J Adolesc Health 2015; 57: 19-23
  • 30 Sinha-Hikim I, Duran P, Shen R, Lee M, Friedman TC, Davidson MB. Effect of long term vitamin D supplementation on biomarkers of inflammation in Latino and African-American subjects with pre-diabetes and hypovitaminosis D. Horm Metab Res 2015; 47: 280-283
  • 31 Kim M, Kim H, Sohn C. Relationship between vitamin K status, bone mineral density, and hs-CRP in young Korean women. Nutr Res Pract 2010; 4: 507-514
  • 32 Mustafar RB, Mohd R, Miswan NA, Bain A, Cader R, Gafor AH, Mohammad M, Shah SA, Kamaruddin NA, Kong NC. The effects of calcitriol with calcium carbonate supplementation on inflammatory biomarkers in chronic kidney disease patients’ with low vitamin D. Cent Eur J Immunol 2014; 39: 236-242
  • 33 Brandi L. 1alpha(OH)D3 One-alpha-hydroxy-cholecalciferol–an active vitamin D analog. Clinical studies on prophylaxis and treatment of secondary hyperparathyroidism in uremic patients on chronic dialysis. Dan Med Bull 2008; 55: 186-210
  • 34 Ohsaki Y, Shirakawa H, Hiwatashi K, Furukawa Y, Mizutani T, Komai M. Vitamin K suppresses lipopolysaccharide-induced inflammation in the rat. Biosci Biotechnol Biochem 2006; 70: 926-932
  • 35 Ohsaki Y, Shirakawa H, Miura A, Giriwono PE, Sato S, Ohashi A, Iribe M, Goto T, Komai M. Vitamin K suppresses the lipopolysaccharide-induced expression of inflammatory cytokines in cultured macrophage-like cells via the inhibition of the activation of nuclear factor kappaB through the repression of IKKalpha/beta phosphorylation. J Nutr Biochem 2010; 21: 1120-1126
  • 36 Asemi Z, Karamali M, Esmaillzadeh A. Effects of calcium-vitamin D co-supplementation on glycaemic control, inflammation and oxidative stress in gestational diabetes: a randomised placebo-controlled trial. Diabetologia 2014; 57: 1798-1806
  • 37 Karamali M, Beihaghi E, Mohammadi AA, Asemi Z. Effects of High-Dose Vitamin D Supplementation on Metabolic Status and Pregnancy Outcomes in Pregnant Women at Risk for Pre-Eclampsia. Horm Metab Res 2015; 47: 867-872
  • 38 de Medeiros Cavalcante IG, Silva AS, Costa MJ, Persuhn DC, Issa CI, de Luna Freire TL, da Conceicao Rodrigues Goncalves M. Effect of vitamin D3 supplementation and influence of BsmI polymorphism of the VDR gene of the inflammatory profile and oxidative stress in elderly women with vitamin D insufficiency: Vitamin D3 megadose reduces inflammatory markers. Exp Gerontol 2015; 66: 10-16
  • 39 Samimi M, Kashi M, Foroozanfard F, Karamali M, Bahmani F, Asemi Z, Hamidian Y, Talari HR, Esmaillzadeh A. The effects of vitamin D plus calcium supplementation on metabolic profiles, biomarkers of inflammation, oxidative stress and pregnancy outcomes in pregnant women at risk for pre-eclampsia. J Hum Nutr Diet. 2015 Oct 14 [Epub ahead of print]
  • 40 Jain SK, Micinski D. Vitamin D upregulates glutamate cysteine ligase and glutathione reductase, and GSH formation, and decreases ROS and MCP-1 and IL-8 secretion in high-glucose exposed U937 monocytes. Biochem Biophys Res Commun 2013; 437: 7-11
  • 41 Ermak G, Davies KJ. Calcium and oxidative stress: from cell signaling to cell death. Mol Immunol 2002; 38: 713-721
  • 42 Tasatargil A, Cadir B, Dalaklioglu S, Yurdakonar E, Caglar S, Turkay C. Effects of vitamin K1 supplementation on vascular responsiveness and oxidative stress in a rat femoral osteotomy model. Cell Biochem Funct 2007; 25: 485-490