Evaluating the effect of arachidonic acid and eicosapentaenoic acid on induction of adipogenesis in human adipose-derived stem cells

Document Type: Original Article


1 Department of Clinical Biochemistry, School of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran

2 Regenerative Medicine Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran

3 Department of Clinical Biochemistry, Faculty of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran


Objective(s): Adipose tissue is one of the most important endocrine organs that liberates many metabolic mediators such as hormones, cytokines, and chemokines. Different types of fatty acids have key roles in adipogenesis. The aim of this study was to evaluate the effects of two essential fatty acids, including Arachidonic acid (AA) and Eicosapentaenoic acid (EPA), on the process of adipogenicity in human Adipose-Derived Stem Cells (hADSCs).
Materials and Methods: After immunophenotyping of hADSCs by flowcytometry, they were differentiated into adipocytes and simultaneously exposed to 30 μM and 60 μM of AA and 25 μM and 50 μM of EPA. Further, along with the MTS assay, the activity of glycalaldehyde-3-phosphate dehydrogenase (GAPDH) was also measured. In addition, expression of lipid markers including peroxisome proliferator-activated receptor γ2 (PPARγ2) and glucose transporter 4 (GLUT4) was evaluated, and the neutral lipid contents were determined using Oil red O staining.
Results: MTS evaluation showed a significant decrease in proliferation in all treatment groups compared to the control group. Based on oil red O staining, fat droplets in the AA treatment groups were higher than in controls. The expression of PPARγ2 and GLUT4 genes and proteins increased in almost all AA and EPA groups compared to control. In addition, GAPDH activity was higher in AA groups than in the control group. In general, while different concentrations of EPA did not increase the adipogenic process compared to the control group, stimulation of differentiation to adipocytes was largely determined by the AA.
Conclusion: The result indicates a positive effect of omega-6 versus omega-3 in stimulating the pathways of adipogenesis.


1.    Klöting N, Blüher M. Adipocyte dysfunction, inflammation and metabolic syndrome. Rev Endocr Metab Dis 2014; 15:277-287.
2.    Qian SW, Li X, Zhang YY, Huang HY, Liu Y, Sun X, et al. Characterization of adipocyte differentiation from human mesenchymal stem cells in bone marrow. BMC Dev Biol 2010; 10:1-11.
3.    McDougald OA, Lane MD. Transcriptional regulation of gene expression during adipocyte differentiation. Annu Rev Biochem 1995; 64:345-373.
4.    Kliewer SA, Sundseth SS, Jones SA, Brown PJ, Wisely GB, Koble CS, et al. Fatty acids and eicosanoids regulate gene expression through direct interactions with peroxisome proliferator-activated receptors α and γ. Proc Natl Acad Sci 1997; 94:4318-4323.
5.    Lehmann JM, Moore LB, Smith-Oliver TA, Wilkison WO, Willson TM, Kliewer SA. An antidiabetic thiazolidinedione is a high affinity ligand for peroxisome proliferator-activated receptor γ (PPARγ). J Biol Chem 1995; 270:12953-12956.
6.    Brodie AE, Manning VA, Ferguson KR, Jewell DE, Hu CY. Conjugated linoleic acid inhibits differentiation of pre-and post-confluent 3T3-L1 preadipocytes but inhibits cell proliferation only in preconfluent cells. J Nutr 1999; 129:602-606.
7.    Kang K, Liu W, Albright KJ, Park Y, Pariza MW. Trans-10, cis-12 CLA inhibits differentiation of 3T3-L1 adipocytes and decreases PPARγ expression. Biochem Bioph Res Co 2003; 303:795-799.
8.    Wang YC, Kuo WH, Chen CY, Lin HY, Wu HT, Liu BH, et al. Docosahexaenoic acid regulates serum amyloid A protein to promote lipolysis through down regulation of perilipin. J Nutr Biochem 2010; 21:317-324.
9.    Kim HK, Della-Fera M, Lin J, Baile CA. Docosahexaenoic acid inhibits adipocyte differentiation and induces apoptosis in 3T3-L1 preadipocytes. J Nutr 2006; 136:2965-2969.
10.    Oster RT, Tishinsky JM, Yuan Z, Robinson LE. Docosahexaenoic acid increases cellular adiponectin mRNA and secreted adiponectin protein, as well as PPARγ mRNA, in 3T3-L1 adipocytes. Appl Physiol Nutr Metab 2010; 35:783-789.
11.    Nikolopoulou E, Papacleovoulou G, Jean-Alphonse F, Grimaldi G, Parker MG, Hanyaloglu AC, et al. Arachidonic acid-dependent gene regulation during preadipocyte differentiation controls adipocyte potential. J lipid Res 2014; 55:2479-2490.
12.    Yin H, Zhou Y, Zhu M, Hou S, Li Z, Zhong H, et al. Role of mitochondria in programmed cell death mediated by arachidonic acid-derived eicosanoids. Mitochondrion 2013; 13:209-224.
13.    Van den Berg JF, Neven AK, Tulen JH, Hofman A, Witteman JC, Miedema HM, et al. Actigraphic sleep duration and fragmentation are related to obesity in the elderly: the Rotterdam Study. Int J Obes 2008; 32:1083-1090.
14.    Goudarzi F, Sarveazad A, Mahmoudi M, Mohammadalipour A, Chahardoli R, Malekshah OM, et al. Combined effect of retinoic acid and calcium on the in vitro differentiation of human adipose-derived stem cells to adipocytes. Arch Physiol Biochem 2018; 124:109-118.
15.    Malekshah OM, Bahrami AR, Afshari JT, Behravan J. PXR and NF-κB correlate with the inducing effects of IL-1β and TNF-α on ABCG2 expression in breast cancer cell lines. Eur J Pharm Sci 2012; 47:474-480.
16.    Qadhi R, Alsaleh N, Samokhvalov V, El-Sikhry H, Bellenger J, Seubert JM. Differential responses to docosahexaenoic acid in primary and immortalized cardiac cells. Toxicol Lett 2013; 219:288-297.
17.    Shaikh SR, Jolly CA, Chapkin RS. n− 3 Polyunsaturated fatty acids exert immunomodulatory effects on lymphocytes by targeting plasma membrane molecular organization. Mol Aspect Med 2012; 33:46-54.
18.    Järvinen R, Tuppurainen M, Erkkilä AT, Penttinen P, Kärkkäinen M, Salovaara K, et al. Associations of dietary polyunsaturated fatty acids with bone mineral density in elderly women. Eur J Clin Nutr 2012; 66:496-503.
19.    Blanchard H, Pédrono F, Boulier-Monthean N, Catheline D, Rioux V, Legrand P. Comparative effects of well-balanced diets enriched in α-linolenic or linoleic acids on LC-PUFA metabolism in rat tissues. Prostaglandins Leukot Essent Fatty Acids 2013; 88:383-389.
20.    Duan Y, Li F, Li L, Fan J, Sun X, Yin Y. n-6: n-3 PUFA ratio is involved in regulating lipid metabolism and inflammation in pigs. Br J Nutr 2014; 111:445-451.
21.    Shirota T, Haji S, Yamasaki M, Iwasaki T, Hidaka T, Takeyama Y, et al. Apoptosis in human pancreatic cancer cells induced by eicosapentaenoic acid. Nutr 2005; 21:1010-1017.
22.    Di Nunzio M, Valli V, Bordoni A. Pro-and anti-oxidant effects of polyunsaturated fatty acid supplementation in HepG2 cells. Prostaglandins Leukot Essent Fatty Acids 2011; 85:121-127.
23.    Grimsgaard S, Bonaa KH, Hansen JB, Nordøy A. Highly purified eicosapentaenoic acid and docosahexaenoic acid in humans have similar triacylglycerol-lowering effects but divergent effects on serum fatty acids. Am J Clin Nutr 1997; 66:649-659.
24.    Casado-Diaz A, Santiago-Mora R, Dorado G, Quesada-Gomez JM. The omega-6 arachidonic fatty acid, but not the omega-3 fatty acids, inhibits osteoblastogenesis and induces adipogenesis of human mesenchymal stem cells: potential implication in osteoporosis. Osteoporos Int 2013; 24:1647-1661.
25.    Wójcik C, Lohe K, Kuang C, Xiao Y, Jouni Z, Poels E. Modulation of adipocyte differentiation by omega‐3 polyunsaturated fatty acids involves the ubiquitin‐proteasome system. J Cell Mol Med 2014; 18:590-599.
26.    Tsuboyama-Kasaoka N, Takahashi M, Kim H, Ezaki O. Up-regulation of liver uncoupling protein-2 mRNA by either fish oil feeding or fibrate administration in mice. Biochem Bioph Res Co 1999; 257:879-885.
27.    Casado-Díaz A, Ferreiro-Vera C, Priego-Capote F, Dorado G, Luque-de-Castro MD, Quesada-Gómez JM. Effects of arachidonic acid on the concentration of hydroxyeicosatetraenoic acids in culture media of mesenchymal stromal cells differentiating into adipocytes or osteoblasts. Genes Nutr 2014; 9:375-389.
28.    Petersen RK, Jørgensen C, Rustan AC, Frøyland L, Muller-Decker K, Furstenberger G, et al. Arachidonic acid-dependent inhibition of adipocyte differentiation requires PKA activity and is associated with sustained expression of cyclooxygenases. J lipid Res 2003; 44:2320-2330.
29.    Madsen L, Petersen RK, Kristiansen K. Regulation of adipocyte differentiation and function by polyunsaturated fatty acids. BBA-Mol Basis Dis 2005; 1740:266-286.
30.    Takahashi Y, Ide T. Dietary n-3 fatty acids affect mRNA level of brown adipose tissue uncoupling protein 1, and white adipose tissue leptin and glucose transporter 4 in the rat. Br J Nutr 2000; 84:175-184.