생명과학회지 (Journal of Life Science)

  • 제21권5호
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  • Pages.720-728
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  • 2011
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
권호 표지
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흑마늘 추출물에 의한 3T3-L1 지방전구세포의 분화 및 adipogenesis 억제에 관한 연구

Inhibition of Adipocyte Differentiation and Adipogenesis by Aged Black Garlic Extracts in 3T3-L1 Preadipocytes

  • 박정애 (동의대학교 블루바이오 소재 개발 센터) ;
  • 박철 (동의대학교 블루바이오 소재 개발 센터) ;
  • 한민호 (동의대학교 대학원 바이오물질제어학과) ;
  • 김병우 (동의대학교 블루바이오 소재 개발 센터) ;
  • 정윤호 (덕산비엔에프주식회사) ;
  • 최영현 (동의대학교 블루바이오 소재 개발 센터)
  • Park, Jung-Ae (Blue-Bio Industry Regional Innovation Center, Dong-Eui University) ;
  • Park, Cheol (Blue-Bio Industry Regional Innovation Center, Dong-Eui University) ;
  • Han, Min-Ho (Department of Biomaterial Control, Graduate School, Dong-Eui University) ;
  • Kim, Byung-Woo (Blue-Bio Industry Regional Innovation Center, Dong-Eui University) ;
  • Chung, Yoon-Ho (Duksan B&F Co. LTD.) ;
  • Choi, Yung-Hyun (Blue-Bio Industry Regional Innovation Center, Dong-Eui University)
  • 투고 : 2011.02.07
  • 심사 : 2011.02.12
  • 발행 : 2011.05.30
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초록

본 연구에서는 3T3-L1 지방전구세포에서 생마늘 및 흑마늘 열수추출물이 insulin, dexamethasone 및 IBMX 등과 같은 분화유도인자에 의하여 유발되는 lipid droplet 및 triglyceride 생성 등과 같은 지방세포로의 분화와 함께 이러한 분화과정에 중요한 역할을 하는 adipogenic transcription factor인 PPAR${\gamma}$, C/EBP${\alpha}$ 및 C/EBP${\beta}$의 발현에 어떠한 영향을 미치는지를 조사하였다. 이를 위하여 성숙한 지방세포에서 나타나는 lipid droplet의 생성에 어떠한 영향을 미치는 지를 확인한 결과, WEABG 처리군에서 lipid droplet의 생성이 현저하게 억제되는 것으로 나타났다. Lipid droplet과 함께 성숙한 지방세포에서 나타나는 특징 중 하나로서 비만의 원인으로 작용하는 지방세포 내의 저장 지방인 triglyceride 생성 정도를 확인한 결과에서도 WEABG 처리군에서 triglyceride 억제효과가 강하게 나타났다. 또한 WERG 및 WEABG-A는 분화유도인자에 의하여 유발된 PPAR${\gamma}$, C/EBP${\alpha}$ 및 C/EBP${\beta}$의 발현 억제에는 큰 영향을 미치지 못하였지만 WEABG는 그들의 발현을 전사 및 번역 수준에서 현저하게 억제시켰다. 이는 WEABG가 adipogenic transcription factor의 발현을 효과적으로 억제함으로서 lipid droplet 및 triglyceride 생성을 억제하여 지방세포로의 분화를 막는 항비만 효능을 가질 수 있음을 의미하는 결과이다. 본 연구 결과는 흑마늘의 비만억제 가능성을 제시하는 것으로서 항비만 기전에 대한 생화학적 해석 및 이를 활용한 향후 지속적인 연구를 위한 귀중한 자료로서 그 가치가 매우 높을 것으로 생각된다.

Garlic (Allium sativum) has been used as a source food as well as a traditional folk medicine ingredient since ancient times. Aged black garlic is a type of fermented garlic and is expected to have stronger anticancer and antioxidant activities than raw garlic. However, the mechanisms of their inhibitory effects on adipocyte differentiation and adipogenesis are poorly understood. In the present study, the effects and mechanisms of water extracts of raw garlic (WERG) and aged black garlic (WEABG) on adipocyte differentiation and adipogenesis in 3T3-L1 preadipocytes were investigated. Treatment with WEABG significantly suppressed terminal differentiation of 3T3-L1 preadipocytes in a dose-dependent manner as confirmed by a decrease in lipid droplet number and lipid content through Oil Red O staining, however WERG had no such effect. In addition, WEABG reduced accumulation of cellular triglyceride, which is associated with a significant inhibition of key pro-adipogenic transcription factors including peroxisome proliferator-activated receptor ${\gamma}$ (PPAR${\gamma}$), cytidine-cytidine-adenosine-adenosine-thymidine (CCAAT)/enhancer binding proteins ${\alpha}$ (C/EBP${\alpha}$) and C/EBP${\beta}$. Taken together, these results provide important new insight that aged black garlic might inhibit adipogenesis by suppressing the pro-adipogenic transcription factors in 3T3-L1 preadipocytes, and further studies will be needed to identify the active compounds that confer the anti-obesity activity of aged black garlic.

키워드

참고문헌

  1. Ahmad, M. S., M. Pischetsrieder, and N. Ahmed. 2007. Aged garlic extract and S-allyl cysteine prevent formation of advanced glycation endproducts. Eur. J. Pharmacol. 561, 32-38. https://doi.org/10.1016/j.ejphar.2007.01.041
  2. Ahmadian, M., Y. Wang, and H. S. Sul. 2010. Lipolysis in adipocytes. Int. J. Biochem. Cell Biol. 42, 555-559. https://doi.org/10.1016/j.biocel.2009.12.009
  3. Alessi, M. C., H. R. Lijnen, D. Bastelica, and I. Juhan-Vague. 2003. Adipose tissue and atherothrombosis. Pathophysiol. Haemost. Thromb. 33, 290-297. https://doi.org/10.1159/000083816
  4. Augusti, K. T. 1996. Therapeutic values of onion (Allium cepa L.) and garlic (Allium sativum L.). Indian. J. Exp. Biol. 34, 634-640.
  5. Bernlohr, D. A., M. A. Bolanowski, T. J. Kelly Jr, and M. D. Lane. 1985. Evidence for an increase in transcription of specific mRNAs during differentiation of 3T3-L1 preadipocytes. J. Biol. Chem. 260, 5563-5567.
  6. Bordia, T., N. Mohammed, M. Thomson, and M. Ali. 1996. An evaluation of garlic and onion as antithrombotic agents. Prostaglandins Leukot. Essent. Fatty Acids 54, 183-186. https://doi.org/10.1016/S0952-3278(96)90014-9
  7. Canizares, P., I. Gracia, L. A. Gomez, C. Martin de Argila, L. de Rafael L, and A. Garcia. 2002. Optimization of Allium sativum solvent extraction for the inhibition of in vitro growth of Helicobacter pylori. Biotechnol. Prog. 18, 1227-1232. https://doi.org/10.1021/bp025592z
  8. Chen, H. C., and R. V. Farese Jr. 2000. DGAT and triglyceride synthesis: a new target for obesity treatment? Trends Cardiovasc. Med. 10, 188-192. https://doi.org/10.1016/S1050-1738(00)00066-9
  9. Cowherd, R. M., R. E. Lyle, and R. E. McGehee Jr. 1999. Molecular regulation of adipocyte differentiation. Semin. Cell Dev. Biol. 10, 3-10. https://doi.org/10.1006/scdb.1998.0276
  10. Duane, W. C. 1997. Cholesterol metabolism in familial hypertriglyceridemia: effects of obesity versus triglyceride level. J. Lab. Clin. Med. 130, 635-642. https://doi.org/10.1016/S0022-2143(97)90113-X
  11. El-Demerdash, F. M., M. I. Yousef, and N. I. El-Naga. 2005. Biochemical study on the hypoglycemic effects of onion and garlic in alloxan-induced diabetic rats. Food Chem. Toxicol. 43, 57-63. https://doi.org/10.1016/j.fct.2004.08.012
  12. El-Jack, A. K., J. K. Hamm, P. F. Pilch, and S. R. Farmer. 1999. Reconstitution of insulin-sensitive glucose transport in fibroblasts requires expression of both PPARgamma and C/EBPalpha. J. Biol. Chem. 274, 7946-7951. https://doi.org/10.1074/jbc.274.12.7946
  13. Farese, R. V. Jr, and T. C. Walther. 2009. Lipid droplets finally get a little R-E-S-P-E-C-T. Cell 139, 855-860. https://doi.org/10.1016/j.cell.2009.11.005
  14. Farmer, S. R. 2006. Transcriptional control of adipocyte formation. Cell Metab. 4, 263-273. https://doi.org/10.1016/j.cmet.2006.07.001
  15. Fox, K. E., D. M. Fankell, P. F. Erickson, S. M. Majka, J. T. Crossno Jr, and D. J. Klemm. 2006. Depletion of cAMP-response element-binding protein/ATF1 inhibits adipogenic conversion of 3T3-L1 cells ectopically expressing CCAAT/enhancer-binding protein (C/EBP) alpha, C/EBP beta, or PPAR gamma 2. J. Biol. Chem. 281, 40341-40353. https://doi.org/10.1074/jbc.M605077200
  16. Freytag, S. O., D. L. Paielli, and J. D. Gilbert. 1994. Ectopic expression of the CCAAT/enhancer-binding protein alpha promotes the adipogenic program in a variety of mouse fibroblastic cells. Genes Dev. 8, 1654-1663. https://doi.org/10.1101/gad.8.14.1654
  17. Galic, S., J. S. Oakhill, and G. R. Steinberg. 2010. Adipose tissue as an endocrine organ. Mol. Cell Endocrinol. 316, 129-139. https://doi.org/10.1016/j.mce.2009.08.018
  18. Gesta, S., Y. H. Tseng, and C. R. Kahn. 2007. Developmental origin of fat: tracking obesity to its source. Cell 131, 242-256. https://doi.org/10.1016/j.cell.2007.10.004
  19. Green, H. and O. Kehinde. 1976. Spontaneous heritable changes leading to increased adipose conversion in 3T3 cells. Cell 7, 105-113. https://doi.org/10.1016/0092-8674(76)90260-9
  20. Green, H. and M. Meuth. 1974. An established pre-adipose cell line and its differentiation in culture. Cell 3, 127-133. https://doi.org/10.1016/0092-8674(74)90116-0
  21. Hafiz, M. S., M. I. Arslan, S. Banarjee, and M. Obaidullah. 2004. Studies on serum triglyceride level in patients with myocardial infarction. Mymensingh. Med. J. 13, 185-187.
  22. Hamm, J. K., B. H. Park, and S. R. Farmer. 2001. A role for C/EBPbeta in regulating peroxisome proliferatoractivated receptor gamma activity during adipogenesis in 3T3-L1 preadipocytes. J. Biol. Chem. 276, 18464-18471. https://doi.org/10.1074/jbc.M100797200
  23. Heilbronn, L., S. R. Smith, and E. Ravussin. 2004. Failure of fat cell proliferation, mitochondrial function and fat oxidation results in ectopic fat storage, insulin resistance and type II diabetes mellitus. Int. J. Obes. Relat. Metab. Disord. 28, S12-21. https://doi.org/10.1038/sj.ijo.0802853
  24. Hodge, G., S. Hodge, and P. Han. 2002. Allium sativum (garlic) suppresses leukocyte inflammatory cytokine production in vitro: potential therapeutic use in the treatment of inflammatory bowel disease. Cytometry 48, 209-215. https://doi.org/10.1002/cyto.10133
  25. Jessen, B. A. and G. J. Stevens. 2002. Expression profiling during adipocyte differentiation of 3T3-L1 fibroblasts. Gene 299, 95-100. https://doi.org/10.1016/S0378-1119(02)01017-X
  26. Kumaraguruparan, R., K. V. Chandra Mohan, S. K. Abraham, and S. Nagini. 2005. Attenuation of N-methyl- N'-nitro-N-nitrosoguanidine induced genotoxicity and oxidative stress by tomato and garlic combination. Life Sci. 76, 2247-2255. https://doi.org/10.1016/j.lfs.2004.11.006
  27. Labreuche, J., P. J. Touboul, and P. Amarenco. 2009. Plasma triglyceride levels and risk of stroke and carotid atherosclerosis: a systematic review of the epidemiological studies. Atherosclerosis 203, 331-345. https://doi.org/10.1016/j.atherosclerosis.2008.08.040
  28. Lafontan, M. and D. Langin. 2009. Lipolysis and lipid mobilization in human adipose tissue. Prog. Lipid Res. 48, 275-297. https://doi.org/10.1016/j.plipres.2009.05.001
  29. Lee, Y. M., O. C. Gweon, Y. J. Seo, J. Im, M. J. Kang, M. J. Kim, and J. I. Kim. 2009. Antioxidant effect of garlic and aged black garlic in animal model of type 2 diabetes mellitus. Nutr. Res. Pract. 3, 156-161. https://doi.org/10.4162/nrp.2009.3.2.156
  30. Manickam, E., A. J. Sinclair, and D. Cameron-Smith. 2010. Suppressive actions of eicosapentaenoic acid on lipid droplet formation in 3T3-L1 adipocytes. Lipids Health Dis. 9, 57. https://doi.org/10.1186/1476-511X-9-57
  31. Patya, M., M. A. Zahalka, A. Vanichkin, A. Rabinkov, T. Miron, D. Mirelman, M. Wilchek, H. M. Lander, and A. Novogrodsky. 2004. Allicin stimulates lymphocytes and elicits an antitumor effect: a possible role of p21ras. Int. Immunol. 16, 275-281. https://doi.org/10.1093/intimm/dxh038
  32. Qidwai, W., R. Qureshi, S. H. Hasan, and S. I. Azam. 2000. Effect of dietary garlic (Allium Sativum) on the blood pressure in humans-a pilot study. J. Pak. Med. Assoc. 50, 204-207.
  33. Rivlin, R. S. 2001. Historical perspective on the use of garlic. J. Nutr. 131, 951S-954S.
  34. Roncari, D. A., D. C. Lau, and S. Kindler. 1981. Exaggerated replication in culture of adipocyte precursors from massively obese persons. Metabolism 30, 425-427. https://doi.org/10.1016/0026-0495(81)90174-8
  35. Rosen, E. D. and O. A. MacDougald. 2006. Adipocyte differentiation from the inside out. Nat. Rev. Mol. Cell Biol. 7, 885-896. https://doi.org/10.1038/nrm2066
  36. Rosen, E. D. and B. M. Spiegelman. 2000. Molecular regulation of adipogenesis. Annu. Rev. Cell Dev. Biol. 16, 145-171. https://doi.org/10.1146/annurev.cellbio.16.1.145
  37. Saltiel, A. R. and C. R. Kahn. 2001. Insulin signalling and the regulation of glucose and lipid metabolism. Nature 414, 799-806. https://doi.org/10.1038/414799a
  38. Schoonjans, K., B. Staels, and J. Auwerx. 1996. The peroxisome proliferator activated receptors (PPARS) and their effects on lipid metabolism and adipocyte differentiation. Biochim. Biophys. Acta 1302, 93-109. https://doi.org/10.1016/0005-2760(96)00066-5
  39. Spiegelman, B. M. and J. S. Flier. 2001. Obesity and the regulation of energy balance. Cell 104, 531-543. https://doi.org/10.1016/S0092-8674(01)00240-9
  40. Thomson, M. and M. Ali. 2003. Garlic [Allium sativum]: a review of its potential use as an anti-cancer agent. Curr. Cancer Drug Targets 3, 67-81. https://doi.org/10.2174/1568009033333736
  41. Vazquez-Vela, M. E., N. Torres, and A. R. Tovar. 2008. White adipose tissue as endocrine organ and its role in obesity. Arch. Med. Res. 39, 715-728. https://doi.org/10.1016/j.arcmed.2008.09.005
  42. Wang, N. D., M. J. Finegold, A. Bradley, C. N. Ou, S. V. Abdelsayed, M. D. Wilde, L. R. Taylor, D. R. Wilson, and G. I. Darlington. 1995. Impaired energy homeostasis in C/EBP alpha knockout mice. Science 269, 1108-1112. https://doi.org/10.1126/science.7652557
  43. Wellman, N. S. and B. Friedberg. 2002. Causes and consequences of adult obesity: health, social and economic impacts in the United States. Asia Pac. J. Clin. Nutr. 11, S705-709. https://doi.org/10.1046/j.1440-6047.11.s8.6.x
  44. White, U. A. and J. M. Stephens. 2010. Transcriptional factors that promote formation of white adipose tissue. Mol. Cell Endocrinol. 318, 10-14. https://doi.org/10.1016/j.mce.2009.08.023
  45. Wu, Z., P. Puigserver, and B. M. Spiegelman. 1999. Transcriptional activation of adipogenesis. Curr. Opin. Cell Biol. 11, 689-694. https://doi.org/10.1016/S0955-0674(99)00037-X
  46. Wyatt, S. B., K. P. Winters, and P. M. Dubbert. 2006. Overweight and obesity: prevalence, consequences, and causes of a growing public health problem. Am. J. Med. Sci. 331, 166-174. https://doi.org/10.1097/00000441-200604000-00002
  47. Yano, T., S. Kobori, M. Sakai, Y. Anami, T. Matsumura, H. Matsuda, M. Kasho, and M. Shichiri. 1997. Beta-very low density lipoprotein induces triglyceride accumulation through receptor mediated endocytotic pathway in 3T3-L1 adipocytes. Atherosclerosis 135, 57-64. https://doi.org/10.1016/S0021-9150(97)00146-9
  48. Yeh, W. C., Z. Cao, M. Classon, and S. L. McKnight. 1995. Cascade regulation of terminal adipocyte differentiation by three members of the C/EBP family of leucine zipper proteins. Genes Dev. 9, 168-181. https://doi.org/10.1101/gad.9.2.168
  49. Yeh, Y. Y. and S. M. Yeh. 1994. Garlic reduces plasma lipids by inhibiting hepatic cholesterol and triacylglycerol synthesis. Lipids 29, 189-193. https://doi.org/10.1007/BF02536728
  50. Zimmermann, R., J. G. Strauss, G. Haemmerle, G. Schoiswohl, R. Birner-Gruenberger, M. Riederer, A. Lass, G. Neuberger, F. Eisenhaber, A. Hermetter, and R. Zechner. 2004. Fat mobilization in adipose tissue is promoted by adipose triglyceride lipase. Science 306, 1383-1386. https://doi.org/10.1126/science.1100747

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