Osteoprotective effects of kefir fortified with omega-3 and vitamin C in ovariectomized rats
Abstract
Abstract: Nutritional interventions can be valuable for the prevention of postmenopausal osteoporosis. This study aimed to investigate the effects of kefir fortified with omega-3 and vitamin C on the bone and uterus parameters of ovariectomized rats. Seventy-seven female Sprague–Dawley rats were ovariectomized or sham-operated. The ovariectomized rats were assigned to six groups and received 1 ml/day of distilled water (OVX group), milk, kefir, kefir fortified with omega-3 fatty acids (kefir+ω3), kefir fortified with vitamin C (kefir+vit-C) or kefir fortified with omega-3 and vitamin C (kefir+ω3+vit-C) for 12 weeks. The sham group also received 1ml/day of distilled water. Subsequently, bone mineral content (BMC) and bone mineral density (BMD) of various bones were assessed. Femurs and uteri were harvested for bone ash analysis and histopathological examinations, respectively. Sera were analyzed for carboxy-terminal cross-linked telopeptide of type 1 collagen, procollagen type 1 amino-terminal propeptide, calcium, phosphorous, tumor necrosis factor-α (TNF-α) and total antioxidant capacity levels. Ovariectomy resulted in significant reduction in bone density (P<0.05). Kefir+ω3+vit-C significantly improved BMC of lumbar spine (0.699±0.027 g compared with 0.580±0.018 in the OVX group), and kefir, kefir+vit-C and kefir+ω3+vit-C significantly increased BMD of tibia (0.118±0.003 g/cm2, 0.119±0.001 and 0.120±0.004 compared with 0.102±0.005 in the OVX group). Moreover, ovariectomy markedly elevated TNF-α level, which was significantly reversed by kefir+ω3+vit-C. Significant atrophy of the uterus was observed following ovariectomy, although the uterus parameters did not change by any of the interventions. In conclusion, kefir fortified with omega-3 fatty acids and vitamin C may have protective effects against bone loss through suppressing inflammation.
References
1 . An overview and management of osteoporosis. Eur J Rheumatol. 2017;4(1):46–56.
2 . The pathophysiology and treatment of osteoporosis. Clin Ther. 2015;37(8):1837–50.
3 . Animal models for osteoporosis. Eur J Pharmacol. 2015;759:287–94.
4 . Effects of a Lactobacillus casei 393 fermented milk product on bone metabolism in ovariectomised rats. Int Dairy J. 2009;19(11):690–5.
5 . The microbiota and health promoting characteristics of the fermented beverage kefir. Front Microbiol. 2016;7:647.
6 . Milk kefir: nutritional, microbiological and health benefits. Nutr Res Rev. 2017;30(1):82–96.
7 Kefir improves bone mass and microarchitecture in an ovariectomized rat model of postmenopausal osteoporosis. Osteoporos Int. 2015;26(2):589–99.
8 Kefir peptides prevent estrogen deficiency-induced bone loss and modulate the structure of the gut microbiota in ovariectomized mice. Nutrients. 2020;12(11):3432.
9 . Short-term effects of kefir-fermented milk consumption on bone mineral density and bone metabolism in a randomized clinical trial of osteoporotic patients. PLoS One. 2015;10(12):e0144231.
10 Low-dose omega-3 fatty acid and vitamin D for anthropometric, biochemical blood indices and respiratory function. Does it work? Int J Vitam Nutr Res. 2020;90(1–2):67–83.
11 . A systematic review of omega-3 fatty acids and osteoporosis. Br J Nutr. 2012;107(Suppl 2):S253–60.
12 . Fish oil suppresses bone resorption by inhibiting osteoclastogenesis through decreased expression of M-CSF, PU.1, MITF and RANK in ovariectomized rats. Mol Med Rep. 2013;7(6):1896–903.
13 . Dietary fish oil results in a greater bone mass and bone formation indices in aged ovariectomized rats. J Bone Miner Metab. 2008;26(3):241–7.
14 . Dietary n-3 fatty acids decrease osteoclastogenesis and loss of bone mass in ovariectomized mice. J Bone Miner Res. 2003;18(7):1206–16.
15 . Dietary intake of fish, n-3 polyunsaturated fatty acids and risk of hip fracture: A systematic review and meta-analysis on observational studies. Crit Rev Food Sci Nutr. 2019;59(8):1320–33.
16 . Is there a role for vitamin C in preventing osteoporosis and fractures? A review of the potential underlying mechanisms and current epidemiological evidence. Nutr Res Rev. 2014;27(2):268–83.
17 Vitamin C improves liver and renal functions in hypothyroid rats by reducing tissue oxidative injury. Int J Vitam Nutr Res. 2020;90(1–2):84–94.
18 . The roles and mechanisms of actions of vitamin C in bone: new developments. J Bone Miner Res. 2015;30(11):1945–55.
19 Effects of ovariectomy and ascorbic acid supplement on oxidative stress parameters and bone mineral density in rats. The Libyan J Med. 2011;6:5965.
20 . Vitamin C reverses bone loss in an osteopenic rat model of osteoporosis. Int J Vitam Nutr Res. 2018;88(1–2):58–64.
21 Vitamin C prevents hypogonadal bone loss. PLoS One. 2012;7(10):e47058.
22 . Protection of sodium arsenite-induced ovarian toxicity by coadministration of L-ascorbate (vitamin C) in mature wistar strain rat. Arch Environ Contam Toxicol. 2001;41(1):83–9.
23 . Supplementation with vitamins E plus C or soy isoflavones in ovariectomized rats: effect on the activities of Na+, K+-ATPase and cholinesterases. Metab Brain Dis. 2007;22(2):156–71.
24 . Pros and cons of fatty acids in bone biology. Prog Lipid Res. 2015;58:121–45.
25 , Omega 3 fatty acids reduce bone resorption while promoting bone generation in rat apical periodontitis. J Endod. 2017;43(6):970–6.
26 Effect of oral administration of kefir on serum proinflammatory cytokines on 5-FU induced oral mucositis in patients with colorectal cancer. Invest New Drugs. 2008;26(6):567–72.
27 Kefir reduces insulin resistance and inflammatory cytokine expression in an animal model of metabolic syndrome. Food Funct. 2016;7(8):3390–401.
28 . Protective actions of soy isoflavones and n-3 PUFAs on bone mass in ovariectomized rats. J Nutr Biochem. 2005;16(8):479–88.
29 .
Bone marrow and stem cell recruitment . In: Anderson JJBGarner SCKlemmer PJ, editors. Diet, nutrients, and bone health. Boca Raton: CRC Press; 2012. p. 28.30 . The potential of probiotics as a therapy for osteoporosis. Microbiol Spectr. 2017;5(4).
31 . Supportive role of probiotic strains in protecting rats from ovariectomy-induced cortical bone loss. Probiotics Antimicrob Proteins. 2019;11(4):1145–54.
32 . An investigation of the association between omega 3 FA and bone mineral density among older adults: results from the National Health and Nutrition Examination Survey years 2005–2008. Osteoporos Int. 2014;25(3):1033–41.
33 . Detrimental effect of eicosapentaenoic acid supplementation on bone following ovariectomy in rats. Prostaglandins Leukot Essent Fatty Acids. 2006;75(6):419–27.
34 . A new insight to bone turnover: role of omega-3 polyunsaturated fatty acids. Sci World J. 2013;2013:589641.
35 Favorable effect of dietary vitamin C on bone mineral density in postmenopausal women (KNHANES IV, 2009): discrepancies regarding skeletal sites, age, and vitamin D status. Osteoporos Int. 2015;26(9):2329–37.
36 . Consumption of different sources of omega-3 polyunsaturated fatty acids by growing female rats affects long bone mass and microarchitecture. Bone. 2011;49(3):455–62.
37 Tumor necrosis factor alpha suppresses the mesenchymal stem cell osteogenesis promoter miR-21 in estrogen deficiency-induced osteoporosis. J Bone Miner Res. 2013;28(3):559–73.
38 Ascorbate-dependent decrease of the mucosal immune inflammatory response to gliadin in coeliac disease patients. Allergol Immunopathol (Madr). 2012;40(1):3–8.
39 . The role of bone turnover markers in monitoring treatment in postmenopausal osteoporosis. Clin Biochem. 2012;45(12):907–19.
40 . The effect of n-3 fatty acids on bone biomarkers in Iranian postmenopausal osteoporotic women: a randomized clinical trial. Age. 2010;32(2):179.
41 . Effect of supplementation of vitamin C and E on oxidative stress in osteoporosis. Indian J Clin Biochem. 2007;22(2):101–5.
42 Anthocyanin-rich blackcurrant extract attenuates ovariectomy-induced bone loss in mice. J Med Food. 2016;19(4):390–7.
43 . Bovine lactoferrin improves bone status of ovariectomized mice via immune function modulation. bone. 2011;48(5):1028–35.
44 Oral administration of alpha-lipoic acid did not affect lipid peroxidation and antioxidant biomarkers in rheumatoid arthritis patients. Int J Vitam Nutr Res. 2019;89(1–2):13–21.
45 . In vitro measurements and interpretation of total antioxidant capacity. Biochim Biophys Acta. 2014;1840(2):931–4.
46 . N-3 poly-unsaturated fatty acids shift estrogen signaling to inhibit human breast cancer cell growth. PLoS One. 2012;7(12):e52838.
