Extra virgin olive oil in maternal diet increases osteogenic genes expression, but high amounts have deleterious effects on bones in mice offspring at adolescence

Document Type : Original Article


1 Department of Cellular Molecular Nutrition, School of Nutritional Sciences and Dietetics, Tehran University of Medical Sciences, Tehran, Iran

2 Department of Biochemistry and Nutrition, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran

3 Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran

4 Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran

5 Department of Statistics, School of Health, Tehran University of Medical Sciences, Tehran, Iran

6 Department of Sport Physiology, School of Physical Education and Sport Sciences, University of Tehran, Tehran, Iran

7 Department of Nutrition, School of Health, Colorectal Research Center, Rasoul Akram Hospital, Iran University of Medical Sciences, Tehran, Iran


Objective(s): Maternal high-fat diet has been shown to have deleterious effects on the offspring bones. However, there is no study to assess the effects of type and amount of maternal dietary oil in an isocaloric diet, with focus on extra virgin olive oil (EVOO). The objective of the current study was to test the hypothesis that type of maternal dietary oil has more effects than its amount in an isocaloric diet during gestation and lactation on bone genes expression in offspring in adolescence.
Materials and Methods: Virgin female C57BL/6 mice were impregnated and fed either the AIN 93G diet (received 16% of calories as soybean oil, as a control diet, or EVOO) or a high fat AIN 93G diet (received 45% of calories as soybean oil or EVOO) from the time of vaginal plug confirmation until offspring’s weaning.
Results: After adjusting for the amount of oils, osteoprotegerin/ receptor activator of nuclear factor NF-κB ligand (OPG/RANK-L) and OPG expressions were 6.1- and 2.8-folds higher in offspring born to EVOO compared with soybean oil-fed mothers. OPG, beta-catenin, and OPG/RANK-L expression were 88%, 94%, and 70% lower in offspring born to the 45% oil-fed mothers compared with the 16% group. In contrast, peroxisome proliferator-activated receptor gamma-2 (PPARγ2) gene expression was higher in the 45% oil group, adjusted for the types of oil.
Conclusion: Maternal EVOO consumption, but not soybean oil increased osteoblastic gene expression, and high amounts of both oils decreased osteoblastic and increased adipogenic genes expression in adolescent offspring.


1. Kanis JA, Burlet N, Cooper C, Rizzoli R, Reginster Y. European guidance for the diagnosis and management of osteoporosis in postmenopausal women. Osteoporos Int 2008; 19:399–428.
2. Goodfellow L, Earl S, Cooper C, Harvey NC. Maternal diet, behaviour and offspring keletal health. Int J Environ Res Public Health 2010; 7:1760–1772.
3. Gluckman PD, Hanson MA, Spencer HG. Predictive adaptive responses and human evolution. Trends Ecol Evol 2005; 20:527–533.
4. Lanham SA, Roberts C, Hollingworth T, Sreekumar R, Elahi MM, Cagampang FR, et al. Maternal high-fat diet: effects on offspring bone structure. Osteoporosis Int 2010; 21:1703-1714.
5. Rodriguez-Bernal CL, Rebagliato M, Iniguez C. Diet quality in early pregnancy and its effects on fetal growth outcomes: the tnfanciay medioambiente (child- hood and environment) mother and child cohort study in Spain. Am J Clin Nutr 2010; 91:1659–1666.
6. Niculescu MD, Lupu DS, Craciunescu CN. Perinatal manipulation of alpha-linolenic acid intake induces epigenetic changes in maternal and offspring livers. FASEB J 2013; 27:350–358.
7. Casado-Díaz A, Santiago-Mora R, Dorado G, Gomez J. The omega-6 arachidonic fatty acid, but not the omega-3 fatty acids, inhibit osteoblastogenesis and induces adipogenesis of human mesenchymal stem cells: potential implication in osteoporosis. Osteoporos Int 2013; 24:1647–1661.
8. Albers J, Keller J, Baranowsky A, Timo Beil F, Catala-Lehnen Ph, Schulze J, Amling M, Schinke Th. Canonical Wnt signaling inhibits osteoclastogenesis independent of osteoprotegerin. J Cell Biol 2013; 4:537-549.
9.  Lecka-Czernik B. PPARγ, an essential regulator of bone mass: metabolic and molecular cues. IBMS Bonekey 2010; 7:171-181.
10. Simopoulos A. Essential fatty acids in health and chronic disease. Am J Clin Nutr 1999; 70:560S–9S.
11. Halade GV, Roohman MM, Williams PJ, Fernandes G. High fat diet-induced animal model of age-associated obesity and osteoporosis. J Nutr Biochem 2010; 21:1162-1169.
12. Cao K, Xu J, Zou X, Li Y, Chen C, Zheng A, et al. Hydroxytyrosol prevents diet-induced metabolic syndromeand attenuates mitochondrial abnormalities in obese mice. Free Radic Biol Med 2014; 67:396–407.
13. Santiago-Mora R, Casado-Díaz A, De Castro MD, Quesada-Gómez JM. Oleuropein enhances osteoblastogenesis and inhibits adipogenesis: the effect on differentiation in stem cells derived from bone marrow. Osteoporos Int 2011; 22:675-684.
14. Gamba CA, Friedman SM, Rodriguez PN, Macri EV, Vacas MI, Lifshitz F. Metabolic status in growing rats fed isocaloric diets with increased carbohydrate-to-fat ratio. Nutrition 2005; 21:249–254.
15. Chen JR, Lazarenko OP, Wu X, Kang J, Blackburn M, Shankar K, et al.  Dietary induced serum phenolic acids promote bone growth via p38 MAPK/beta-catenin canonical Wnt signaling. J Bone Miner Res 2010; 25:2399–2411.
16. Lima Reboucas E, Nascimento Costa J, Passos M, Sousa Passos J, Van den Hurk R, Viana Silva J. Real time PCR and importance of housekeepings genes for normalization and quantification of mRNA expression in different tissues. Braz Arch Biol Technol 2013; 56: 143-154.
17. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods 2001; 25:402-408.
18. Priego T, Sánchez J, García AP, Palou A, Picó C. Maternal dietary fat affects milk fatty acid profile and impacts on weight gain and thermogenic capacity of suckling rats. Lipids 2013; 48:481-495.
19. Rosa BV, Blair H, Vickers MH, Dittmer KE, Morel P, Knight CG, et al. Moderate exercise during pregnancy in wistar rats alters bone and body composition of the adult offspring in a sex-dependent manner. PLoS One 2013; 8:e82378.
20. Wohl GR, Loehrke L, Watkins BA, Zernicke RF. Effects of high fat diet on mature bone mineral content, structure, and mechanical properties. Calcif Tissue Int 1998; 63:74-79.
21. Orchard TS, Cauley JA, Frank GC, L Neuhouser M,  Robinson JG,  Snetselaar L, Tylavsky F, et al. Fatty acid consumption and risk of fracture in the Women’s Health Initiative. Am J Clin Nutr 2010; 92:1452- 1460.
22. Watkins BA, Li Y, Lippman HE, Fengs S. Modulatory effect of omega-3 polyunsaturated fatty acids on osteoblast function and bone metabolism. Prostaglandins Leukot Essenti Fatty Acids 2003; 68:387-398.
23. El-Sayed E, Ibrahim K. Effect of the types of dietary fats and non-dietary oils on bone metabolism. Crit Rev Food Sci Nutr 2015 16:0.
24. Devlin MJ, Grasemann C, Cloutier AM, Louis L, Palmert M, Bouxsein ML. Maternal perinatal diet induces developmental programming of bone architecture. J Endocrinol 2013; 217:69–81.
25. Horowitz MC, Lorenzo JA. The origins of osteoclasts. Curr Opin Rheumatol 2004; 16: 464–468.
26. Gimble JM, Robinson CE, Wu X, Kelly KA. The function of adipocytes in the bone marrow stroma: an update. Bone 1996; 19:421–428.
27. Yasuda H, Shima N, Nakagawa N, Yamaguchi K, Kinosaki M, Mochizuki Sh, et al. Identity of osteoclastogenesis inhibitory factor (OCIF) and osteoprotegerin (OPG): a mechanism by which OPG/OCIF inhibits osteoclastogenesis in vitro. Endocrinology 1998; 39:1329–1337.
28. Fuller K, Wong B, Fox S, Choi Y, Chambers T. TRANCE is necessary and sufficient for osteoblast-mediated activation of bone resorption in osteoclasts. J Exp Med 1998; 188:997–1001.
29. Hofbauer LC, Lacey DL, Dunstan CR, Spelsberg TC, Riggs BL, Khosla S. Interleukin-1β and tumor necrosis factor-α, but not interleukin-6, stimulate osteoprotegerin 
ligand gene expression in human osteoblastic cells. Bone 1999; 25:255–259.
30. Chen J, Lazarenko O, Wu X. Obesity reduces bone density associated with activation of PPARγ and suppression of Wnt/β-catenin in rapidly growing male rats. PLoS One 2010; 5: 13704.
31. Saika M, Inoue D, Kido S, Matsumoto T. 17‚-Estradiol stimulates expression of osteoprotegerin by a mouse stromal cell line, ST-2, via estrogen receptor. Endocrinology 2001; 142:2205-2212.
32. Chen XW, Garner SC, Anderson JJ. Isoflavones regulate interleukin-6 and osteoprotegerin synthesis during osteoblast cell differentiation via an estrogen-receptordependent pathway. Biochem Biophys Res Commun 2002; 295:417-422.
33. Shevde NK, Bendixen AC, Dienger KM, Pike JW. Estrogens suppress RANK ligand-induced osteoclast differentiation via a stromal cell independent mechanism involving c-Jun repression. Proc Natl Acad Sci USA 2000; 97:7829-7834.
34. Srivastava S, Toraldo G, Weitzmann MN, Cenci S, Ross FP, Pacifici R. Estrogen decreases osteoclast formation by down-regulating receptor activator of NF-ÎB ligand (RANKL)-induced JNK activation. J Biol Chem 2001; 276:8836-8840.
35. Hughes DE, Dai A, Tiffee JC, Hiu Li HH, Mundy GR, Boyce BF. Estrogen promotes apoptosis of murine osteoclasts mediated by TGF-β. Nat Med 1996; 2:1132–1136.