The role of probiotic content of kefir in the adipogenic differentiation process of 3T3-L1 cells

Main Article Content

Sabiha Gökçen Zeybek
Eda Becer
Tamer Şanlıdağ
Sevinç Yücecan
Seda Vatansever

Keywords

Kefir, Obesity, Leptin, ANGPTL-4, PPAR-Ɣ

Abstract

Objective: Kefir is traditional fermented milk that is reported to have various health benefits such as preventing obesity. The aim of this study was to investigate the effect of kefir on lipid accumulation and cell differentiation in adipocytes.


Materials and Methods: Kefir was prepared from milk and kefir powder fermentation. Then, kefir supernatant and pellet fractions were incubated for 24 h and 48 h with mature 3T3-L1 adipocytes. Cytotoxic effect of kefir on the cells was measured by MTT assay. Lipid accumulation was detected by Oil-red O staining.  Anti-adipogenic activities of kefir fractions were investigated distribution of ANGPTL-4, leptin and PPAR- γ in mature 3T3-L1 adipocytes using indirect immunoperoxidase technique.


Results: In the MTT assay, 0.1 mg/dl dilutions of kefir supernatant and pellet were found to be effective on adipocytes growth and differentiation after both 24 h and 48 h incubations. Lipid accumulation was reduced by both kefir supernatant and pellet in 3T3-L1 adipocytes for 24 h. As a result of immunohistochemical staining, ANGPTL-4 immunoreactivity was significantly increased in 3T3-L1 cells after treated with kefir fractions. Additionally, leptin immunoreactivity was also decreased significantly in the mature adipocytes after treated with kefir pellet. There were no differences in expression of PPAR- γ in all groups.


Conclusion: Kefir fractions have prevented lipid accumulation by preserving the cell structure on 3T3-L1 adipocytes. The increased level of ANGPTL4 expression may be mediated inhibition of lipid accumulation.  Moreover, kefir supernatant may play important role in preventing weight gain due to reduced leptin level.

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References

1. Sanchez M, Panahi S, Tremblay A. Childhood Obesity: A Role for Gut Microbiota. Int J Environ Res Public Health 2015; 12(1):162 – 175.
2. Campbell AW. The Epidemic of Obesity. Altern Ther Health Med 2018; 24 (1): 8-11.
3. Tremaroli V, Bäckhed F. Functional interactions between the gut microbiota and host metabolism. Nature 2012; 489 (7415): 242-249.
4. Miele L, Giorgio V, Alberelli M, Candia E, Gasbarrini A, Grieco A. Impact of Gut Microbiota on Obesity. Diabetes and Cardiovascular Disease Risk. Curr Cardiol Rep 2015; 17 (12): 120.
5. Cax A, West N, Cripss A. Obesity, inflammation and the gut microbiota. Lancet Diabetes Endocrinol 2015; 3 (3): 207- 215.
6. Marik P. Colonic flora, probiotics, obesity and diabetes. Front Endocrinol 2012; 3 (87): 1-6.
7. Molinaro F, Paschetta E, Cassader M, Gambino R, Musso G. Probiotics, prebiotics energy balance and obesity. Gastroenterol Clin North Am 2012; 41(4): 843- 854.
8. Sanders M, Benson A, Lebeer S, Merenstein D, Klaenhammer T. Shared mechanisms among probiotic taxa: implications for general probiotic claims. Curr Opin Biotechnol 2018; 49: 207-216.
9. Brusaferro A, Cozzali R, Orabona C, et al. Is it time to use probiotics to prevent or treat obesity?. Nutrients 2018;10 (1613): 1-14.
10. Gerard P. Gut Microbiota and Obesity. Cell Mol Life Sci 2016; 73 (1): 147-162.
11. Rosa D, Dias M, Grzeskowiak L, Reis S, Conceicao L, Peluzio M. Milk kefir: nutritional, microbiological and health benefits. Nutr Res Rev 2017; 30 (1): 82-96.
12. Rosa D, Grzeskowiak L, Ferreira C, Fonseca A, Reis SA, Dias MM, et al. Kefir reduces insulin resistance and inflammatory cytokine expression in animal model of the metabolic syndrome. Food Funct 2016; 7 (8): 3390-3401.
13. Khan M, Gerasimidis K, Edwards C, Shaikh G. Role of gut microbiota in the aetiology of obesity proposed mechanisms and review of the literature. J Obes 2016; 2016 (7353642): 1-28.
14. Mazloom K, Siddiqi I, Covasa M. Probiotics: How effective are they in the fight against obesity. Nutrients 2019;11 (258): 1-24.
15. Boulange C, Neves A, Chiloux J, Nicholson J, Dumas M. Impact of the gut microbiota on inflammation, obesity, and metabolic disease. Genom Med 2016; 8 (42): 1-12.
16. Shabana S. Shahid U. The Gut Microbiota and its potential role in obesity. Future Microbiol 2018; 13 (5): 589 – 603.
17. Zebisch K, Voigt V, Wabitsch M, Brandsch M. Protocol for effective differentiation of 3T3-L1 cells to adipocytes. Anal Biochem 2012; 425 (2012): 88-90.
18. Stoecker K, Sass S, Theis F, Hauner H, Pfaafl M. Inhibition of fat cell differentiation in 3T3-L1 pre-adipocytes by all- trans retinoic acid: Integrative analysis of transcriptomic and phenothypic data. Biomol Detect Quantif 2017; 11 (2017): 31 – 44.
19. Paul L, Satheeshkumar N. AMPK activating and Anti adipogenic potential of Hisbiscus rosa sinensis flower in 3T3-L1 cells. J Ethnopharmacol 2018; 26 (233): 123-130.
20. Huang J, Yokoyama W, Kim Y. Soy noodles processed from soy flour or tofu affects antioxidant content, lipid accumulation in 3T3-L1 cells and plasma lipids in hamsters. J Food Process Preserv 2019; 0 (0): 1- 8.
21. Erdogan F, Ozarslan S, Seydim Z, Tas T. The effect of kefir produced from natural kefir grains on the intestinal microbial populations and antioxidant capasities of Balb/c mice. Food Res Int 2019; 115 (2019): 408 – 413.
22. Paglia L, Listi A, Caruso S, Amodeo V, Passiglia F, Bazan V. Potential Role of ANGPTL4 in the cross talk between metabolism and cancer through PPAR signaling pathway. PPAR Res 2017; 2017 (8187235): 1- 15.
23. Kim H, Youn B, Shin M, Namkoong C, Park K, Baik J. Hypothalamic ANGPTL4/FIAF is a novel regulator of food intake and body weight. Diabetes 2010; 59 (11): 2772-2780.
24. Dahiya DK, Renuka M. Puniya UK. Shandilya T. Dhewa N. Kumar, S. Kumar AK. Puniya P. Gut Microbiota Modulation ıts relationship fibers and probiotics: A review. Front Microbiol 2017; 8 (563): 1 – 17.
25. Morris A. ANGPTL4- the link binding obesity and glucose intolerance. Nat Rev Endocrinol 2018; 14 (5): 251.
26. Janssen A, Katiraei S, Bartosinska B, Eberhard D, Dijk K, Kersten S. Loss of angiopoietin-like 4 (ANGPTL4) in mice with diet-induced obesity uncouples visceral obesity from glucose intolerance partly via the gut microbiota, Diabetologia 2018; 61 (6): 1447-1458.
27. Koliwad S, Gray N, Wang J. Angiopoietin-like 4 (Angptl4). Adipocyte 2012; 1 (3): 182-187.
28. D’souza A, Neumann U, Glavas M, Kleffer T. The glucoregulatory actions of leptin. Mol Metab 2017; 6 (9) 1052-1065.
29. Stern J, Rutkowski J, Scherer P. Adiponectin, Leptin and Fatty Acids in the Maintenance of metabolic homeostasis through adipose tissue crosstalk. Cell Metab 2016; 23 (5): 770- 784.
30. Mechanick J, Zhao S, Garvey T. Leptin, An Adipokine with Central Importance in the global obesity problem. Global Heart. 2018; 13 (2): 113-127.
31. Friedman J. The long road to leptin. J Clin Invest 2016; 126 (12): 4727-4734.
32. Gezginc Y, Maranci C. Effect of fermented food consumption on biochemical parameters and adipokines levels. Prog Nutr 2018; 20 (4): 642- 647.
33. Han L, Shen W, Bittner S, Kraemer F, Azhar S. PPARs: regulators of metabolism and as therapeutic targets in cardiovascular disease. Part II: PPAR-β/δand PPAR-γ. Future Cardiol 2017;13 (3): 279 – 296.
34. Janani C, Kumari BD. PPAR gamma gene – A review. Diabetes Metab Syndr 2015; 9 (2015): 46 – 50.
35. Feng S, Reuss L, Wang Y. Potential of Natural Products in the Inhibition of adipogenesis through Regulation of PPAR-γ Expression and/or Its Transcriptional Activity. Molecules 2016; 21 (10): 1278- 1297.
36. Hasan A, Rahman A, Kobori H. Interactions between Host PPARs and Gut Microbiota in Health and Disease. Int J Mol Sci 2019; 20 (2): 387- 406.
37. Sabry M, Dawood A, Rashed L, Sayed S, Hassan S, Yaunes S. Relation between resistin, PPAR-γ, obesity and atherosclerosis in male albino rats. Arch Physiol Biochem 2019; 7:1 – 10.
38. Polvani S, Tarocchi M, Tempesti S, Bencini L, Galli A. Peroxisome proliferator activated receptors at the crossroad of obesity, diabetes, and pancreatic cancer. World J Gastroenterol 2016; 22 (8): 2441 – 2459.
39. Ho J, Choi J, Lim W, Kim M, Lee I, Cho H. Kefir inhibits 3T3-L1 adipocyte differentiation through down-regulation of adipogenic transcription factor expression. J Sci Food Agric 2013; 93 (3): 485 – 90.
40. Choi J, Kang H, Lim W, Kim M, Lee I, Cho H. Kefir prevented excess fat accumulation in dietinduced obese mice. Biosci. Biotechnol Biochem 2017; 81 (5): 958-965.

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