Effects of Ethanol Extract of Sargassum horneri on Adipocyte Differentiation and Adipogenesis in 3T3-L1 Preadipocytes

괭생이모자반 에탄올 추출물이 3T3-L1 지방전구세포의 분화 및 adipogenesis에 미치는 영향

  • 권다혜 (동의대학교 항노화 연구소) ;
  • 최영현 (동의대학교 항노화 연구소) ;
  • 김병우 (블루바이오 소재개발 및 실용화지원센터) ;
  • 황혜진 (블루바이오 소재개발 및 실용화지원센터)
  • Received : 2018.10.11
  • Accepted : 2018.11.14
  • Published : 2019.02.28


Sargassum horneri (Turner) C. Agardh is a marine brown algae widely distributed in the North Pacific Ocean. It is known for its anti-inflammatory and anti-atopic effects. In this study, we determined the effects of ethanol extract of Sargassum horneri (Turner) C. Agardh (EESH) on anti-obesity activities in 3T3-L1 preadipocytes. Our results indicated that treatment with EESH decreased the differentiation of 3T3-L1 preadipocytes in a dose-dependent manner, as confirmed by a decrease in lipid droplet content observed by oil red O staining. The concentrations of cellular triglycerides were also reduced in 3T3-L1 cells after treatment with EESH. Triglyceride content was inhibited by 13%, 16%, and 23% after treatment with 250, 500, and $1,000{\mu}g/ml$ of EESH in 3T3-L1 cells, respectively. Western blotting analysis showed that EESH suppressed adipogenic transcription factor expression in a dose dependent manner. Specifically, it suppressed cytidine-cytidine-adinosine-adenosine-thymidine (CCAAT) /enhancer binding proteins $(C/EBP){\alpha}$, $C/EBP{\beta}$ and peroxisome proliferator-activated receptor $(PPAR){\gamma}$. This indicated that EESH could control the expression of adipogenic transcription factors and inhibit the differentiation of adipocytes. Taken together, these findings demonstrated that EESH showed anti-obesity effects and could have potential uses in the field of nutraceuticals.

본 연구에서는 괭생이모자반 에탄올 추출물의 항비만 효과에 대해 알아보고자 3T3-L1 preadipocyte에서 Oil Red O staining과 triglyceride content, adipogenesis의 발현에 대해 조사하였다. MTT assay를 통해 괭생이모자반 에탄올 추출물의 세포 독성을 확인 해 본 결과 $200{\sim}1,000{\mu}g/ml$ 농도에서 90% 이상의 세포 생존율을 확인 할 수 있었다. 또한 insulin, dexamethasone과 IBMX 복합물을 처리하여 분화를 유도하였을 때는 분화유도제를 처리하지 않은 군에 비해 활발하게 지방구를 형성하였으나, 괭생이모자반 에탄올 추출물을 250, 500, $1,000{\mu}g/ml$ 농도로 처리하였을 때는 농도 의존적으로 지방구 형성을 억제하였다. 이를 정량하여 triglyceride content (%)로 나타낸 결과 농도별로 13%, 16%, 23% 억제 효과를 나타내었다. 또한 Western blot assay를 통해 adipogenic transcription factor의 발현양상을 비교해 본 결과, 분화유도제 처리에 의해 $C/EBP{\alpha}$, $C/EBP{\beta}$$PPAR{\gamma}$의 발현이 증가하였으나, 괭생이모자반 에탄올 추출물을 처리하였을 때는 이러한 전사인자들의 발현이 농도 의존적으로 감소됨을 확인 할 수 있었다. 이상의 결과를 통해 괭생이모자반 에탄올 추출물은 adipogenic transcription factor의 발현을 조절하여 지방세포의 분화를 억제하였으며, 지방구와 triglyceride 생성을 감소시켜 최종적으로 비만억제 효과를 나타낸다는 것을 알 수 있었다.


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Fig. 1. Effects of EESH on the cell viability in 3T3-L1 preadipocytes.

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Fig. 2. Effects of EESH on the phase contrast microscopic morphological changes of differentiated 3T3-L1 preadipocyte.

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Fig. 3. Effects of EESH on the lipid droplet accumulation inhibition of differentiated 3T3-L1 preadipocyte.

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Fig. 4. Effects of EESH on the levels of adipogenic transcription factors protein expression by Western blot analysis in differentiated 3T3-L1 preadipocyte.


  1. Chen, H. C. and Farese, R. V. 2005. Inhibition of triglyceride synthesis as a treatment strategy for obesity : lessons from DGAT1-deficient mice. Arterioscle. Thromb. Vasc. Biol. 25, 482-486.
  2. Chen, Z., Torrens, J. I., Anand, A., Spiegelman, B. M. and Friedman, J. M. 2005. Krox20 stimulates adipogenesis via C/EBPbeta-dependent and -independent mechanisms. Cell Metab. 1, 93-106.
  3. Choi, E. O., Kim, H. S., Han, M. H., Choi, Y. H., Kim, B. W., Hwang, J. A. and Hwang, H. J. 2012. Effects of Hizikia fusiforme extracts on adipocyte differentiation and adipogenesis in 3T3-L1 preadpocytes. J. Life Sci. 22, 1399-1406.
  4. Choi, E. O., Kim, H. S., Han, M. H., Park, C., Choi, Y. H. and Hwang, H. J. 2012. Anti-obesity activities of methyl alcohol extract of Enteromorpha linza in 3T3-L1 preadipocytes. Cancer Prev. Res. 18, 80-87.
  5. Choi, H. J., Kil, J. H., Bak, S. S., Kong, C. S., Park, K. Y., Sep, Y. W. and Lim, S. Y. 2006. Inhibitory effects of solvent extracts from seven brown algae on mutagenicity and growth of human cancer cells. J. Life Sci. 16, 1080-1086.
  6. Fajas, L., Fruchart, J. C. and Auwerx, J. 1998. Transcriptional control of adipogenesis. Curr. Opinion. Cell Biol. 10, 165-173.
  7. Haslam, D. W. and James, W. P. 2005. Obesity. Lancet 366, 1197-1209.
  8. Kim, B. M., Jun, J. Y., Park, Y. B. and Jeong, I. H. 2006. Antioxidative activity of methanolic extracts from seaweeds. J. Kor. Soc. Food. Sci. Nutr. 35, 1097-1101.
  9. Kim, H. S., Kwon, D. H., Cheon, J. M., Choi, E. O., Kim, J. H., Han, M. H., Choi, Y. H., Kim, B. W. and Hwang, H. J. 2015. Effects of Myelophycus Simplex Papenfuss methanol extract on adipocyte differentiation and adipogenesis in 3T3-L1 preadipocytes. J. Life Sci. 25, 62-67.
  10. Kim, M. E., Jung, Y. C., Jung, I. A., Lee, H. W., Youn, H. Y. and Lee, J. S. 2014. Anti-inflammatory effects of ethanolic extract from Sargassum horneri (Turner) C. Agardh on lipopolysaccharide-stimulated macrophage activation via NF-kB pathway regulation. Immunol. Invest. 44, 137-183.
  11. Yu, K. H., Ahn, B. N., Lee, S. H., Kim, M. H., Mun, O. J., Karadeniz, F., Kong, C. S., Kim, Y. Y. and Kwon, M. S. 2014. Sargassum sp. attenuates oxidative stress and suppresses lipid accumulation in vitro. J. Life Sci. 24, 273-283.
  12. Kim, Y. M., Kim, D. S. and Choi, T. S. 2004. Anticoagulant activities of brown seaweed extracts in korea. Kor. J. Food Sci. Technol. 36, 1008-1013.
  13. Lee, C. J. 2010. Inhibitory Effects of Sargassum horneri extracts on atopic disease. M.D. dissertation, Pukyong National University. Busan, Korea.
  14. Leung, W. Y., Thomas, G. N., Chan, J. C. and Tomlinson, B. 2003. Weight management and current options in pharmacotherapy : orlistat and sibutramine. Clin. Ther. 25, 58-80.
  15. Morrison, R. F. and Farmer, S. R. 2000. Hormonal signaling and transcriptional control of adipocyte differentiation. Nutrition 130, 3116S-3121S.
  16. Rosen, E. D. and MacDougald, O. A. 2006. Adipocyte differentiation from the inside out. Nat. Rev. Mol. Cell. Biol. 7, 885-896.
  17. Suh, K. H., Ahn, K. H. and Cho, M. C. 1999. Biosorption of $Pb^{1+}$ and $Cr^{3+}$ by using Sargassum horneri. J. Kor. Environ. Sci. Soc. 8, 387-391.
  18. Wellman, N. S. and Friedberg, B. 2002. Causes and consequences of adult obesity : health, social and economic impacts in the United States. Asia Pac. J. Clin. Nutr. 11, S705-709.
  19. Wu, Z., Bucher, N. L. and Farmer, S. R. 1996. Induction of peroxisome proliferator-activated receptor gamma during the conversion of 3T3 fibroblasts into adipocytes is mediated by C/EBPbeta, C/EBPdelta, and glucocorticoids. Mol. Cell Biol. 16, 4128-4136.
  20. Wu, Z., Xie, Y., Bucher, N. L. and Farmer, S. R. 1995. Conditional ectopic expression of C/EBP beta in NIH-3T3 cells induces PPAR gamma and stimulates adipogenesis. Genes Dev. 9, 2350-2363.
  21. Yeh, W. C., Cao, Z., Classon, M. and Mcknight, S. L. 1995. Cascade regulation of terminal adipocyte differentiation by three members of the C/EBP family of leucine zipper proteins. Genes Dev. 9, 168-181.
  22. Zhang, J. W., Klemm, D. J., Vinson, C. and Lane, M. D. 2004. Role of CREB in transcriptional regulation of CCAAT/enhancer- binding protein beta gene during adipogenesis. J. Biol. Chem. 279, 4471-4478.