Korean Journal of Clinical Laboratory Science (대한임상검사과학회지)

Korean Society for Clinical Laboratory Science (대한임상검사과학회)
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The Lipid Efflux Effects of Dichloromethane Extract from Orostachys japonicus in 3T3-L1 Adipocyte Cells

3T3-L1 지방세포에 대한 와송 디클로로메탄 추출물의 지질 대사 개선에 관한 연구

  • Kim, Soo-Hwan (Department of Biomedical Laboratory Science, Gimcheon University) ;
  • Lee, Hyeong-Seon (Department of Biomedical Laboratory Science, Jungwon University)
  • 김수환 (김천대학교 임상병리학과) ;
  • 이형선 (중원대학교 임상병리학과)
  • Received : 2019.10.10
  • Accepted : 2019.11.26
  • Published : 2019.12.31
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Abstract

This study investigated the improved lipid metabolism effect of 3T3-L1 cells induced by adipocytes using the dichloromethane (DCM) fraction in the organic solvent extract of Wassong (Orostachys japonicus). To confirm the cell cytotoxicity, each of 6 fractions of organic solvent extracts (EtOH, Hexane, DCM, EtOAc, BuOH, and H2O) was examined using MTS assay. As a result, it was confirmed that the DCM extract was stable over the whole range of concentrations, and a DCM fraction was used to confirm the improved lipid metabolism effect. Lipid excretion was measured to confirm the change of lipid metabolism. 3T3-L1 cells induced by adipocytes were treated with DCM extract and stained with oil-red O to evaluate lipid accumulation. As a result, it was confirmed that the lipid efflux was significantly improved. In order to confirm the mechanism of lipid efflux, the mRNA expressions of ABCA1 and ABCG1, which are lipid transport proteins, were confirmed by real-time PCR. Therefore, the present study confirmed that the DCM extract from Orostachys japonicus has the effect of improving the lipid metabolism on 3T3-L1 adipocytes. In addition, the results of this study will be used as the basis for the development of functional foods using Orostachys japonicus and also for conducting research on the detailed mechanisms.

본 연구는 와송 유기용매 추출물중 DCM층을 이용하여 지방세포로 유도된 3T3-L1세포에 대한 지질대사 개선효과를 확인하는 연구이다. 세포독성을 확인하기 위하여 MTS-assay를 이용하여 6가지 분획의 와송 유기용매 추출물(EtOH, Hexane, DCM, EToAc, BuOH, H2O)에 대한 독성검사를 실시하여 세포에 대한 안정성을 확인하였다. 그 결과 DCM추출물이 전 농도에 걸쳐 안정성이 있음을 확인하고, 지질대사 개선효과를 확인하기 위하여 DCM층을 사용하였다. 우선 지질대사 개선효과를 확인하기 위하여 지질 배출능을 측정하였다. 지방세포로 유도된 3T3-L1세포에 대하여 DCM추출물을 처리한 후 oil-red O염색을 실시하여 지질 축적량을 평가한 결과 지질 배출능이 개선된 것을 확인할 수 있었다. 또한 지질 배출능 분석을 위하여 지질수송 단백질인 ABCA1, ABCG1의 mRNA발현을 real-time PCR로 확인한 결과 DCM추출물 처리 시 유의적인 발현 증가가 나타났음을 확인 하였다. 따라서 본 연구를 통하여 와송 DCM추출물이 3T3-L1 지방세포에 대하여 지질대사 개선효과가 있음을 확인하였다.

Keywords

References

  1. Lee JJ, Park MR, Kim AR, Lee MY. Effect of ramie leaves on improvement of lipid metabolism and anti-obesity effect in rats fed a high fat/high cholesterol diet. Kor J Food Sci. Technol. 2011;3:83-90. http://doi.org/10.9721/KJFST.2011.43.1.083
  2. Lim S, Shin H, Song JH, Kwak SH, Kang SM, Yoon JW, et al. Increasing prevalence of metabolic syndrome in Korea. Diabetes Care. 2011;34:1323-1328. https://doi.org/10.2337%2Fdc10-2109 https://doi.org/10.2337/dc10-2109
  3. Ministry of Health & Welfare. Yearbook of health and welfare statistics. Seoul: Report of the Ministry of Health and Welfare; Research report. 2003. p35.
  4. Ministry of Health & Welfare. 2014 National health and nutrition examination survey. Sejong: Korea Centers for Disease Control & Prevention; Research report. 2015. p110-112.
  5. Krauss RM. Triglycerides and atherogenic lipoproteins: rationale for lipid management. Am J Med. 1998;105(1A):S58-62. https://doi.org/10.1016/s0002-9343(98)00213-7
  6. Baumer Y, McCurdy S, Weatherby TM, Mehta NN, Halbherr S, Halbherr P, et al. Hyperlipidemia-induced cholesterol crystal production by endothelial cells promotes atherogenesis. Nat Commun. 2017;8:1129. https://doi.org/10.1038/s41467-017-01186-z
  7. Moon SJ. Nutritional problems of Korean. Kor J Nutr. 1996;29:371-380.
  8. Mckenny JM. Lipid management: tools for getting to the goal. Am J Manage Care. 2001;7(9 Suppl):299-306.
  9. Miettinen TA. Cholesterol absorption inhibition: a strategy for cholesterol-lowering therapy. Int J Clin Pract. 2001;55:710-716.
  10. Shin MK, Han SH. Effects of methanol extracts from bamboo (Pseudosasa japonica Makino) leaves extracts on lipid metabolism in rats fed high fat and high cholesterol diet. Kor J Diet Culture. 2002;17:30-36.
  11. Park JE, Kee H. Cha YS. Effect of Stevia rebaudiana Bertoni leaf extract on antiobesity in C57BL/6J mice. Kor J Food Sci Technol. 2010;42:586-592.
  12. Kim JY, Jung EJ, Won YS, Lee JH, Shin DY, Seo KI. Cultivated Orostachys japonicus induces apoptosis in human colon cancer cells. Kor J Food Sci. Technol. 2012;44:317-323. http://doi.org/10.9721/KJFST.2012.44.3.317
  13. Choi SY, Kim JG, Sung NJ. Studies on the physicochemical chracteristics and NDMA formation of Orostachys japonicus A. berger. J Kor Soc Food Sci. Nutr. 2008;21:148-156.
  14. Park HJ, Young HS, Kim JO, Rhee SH, Choi JS. A study on the chemical constituents of Orostachys japonicus A. berger. Kor J Pharm. 1991;22:78-84.
  15. Park JG, Park JC, Hur JM, Park SJ, Choi DR, Shin DY, et al. Phenolic compounds from Orostachys japonicus having anti-HIV-1 protease activity. Nat Prod Sci. 2000;6:117-121.
  16. Park HJ, Lim SC, Lee MS, Young HS. Triterpene and steroids from Orostachys japonicus. Kor J Pharm. 1994;25:20-24.
  17. Kwon J, Han KS. Effects of Orostachys japonicus A. Berger on the immune system. Kor J Med Crop Sci. 2004;12:315-320.
  18. Choi SY, Chung MJ, Sung NJ. Studies on the antioxidative ability of methanol and water extracts from Orostachys japonicus A. berger according to harvest times. J Kor Soc Food Sci Nutr. 2008;21:157-164.
  19. Lee SJ, Seo JK, Shin JH, Lee HJ, Sung NJ. Antioxidant activity of wa-song (Orostachys japonicus A. Berger) according to drying methods. J. Kor Soc Food Sci Nutr. 2008;37:605-611. http://doi.org/10.3746/jkfn.2008.37.5.605
  20. Yoon SY, Lee SY, Kim KBWR, Song EJ, Kim SJ, Lee SJ, et al. Antimicrobial activity of the solvent extract from different parts of Orostachys japonicus. J Kor Soc Food Sci Nutr. 2009;38:14-18. http://doi.org/10.3746/jkfn.2009.38.1.01
  21. Ryu DS, Baek GO, Kim EY, Kim KH, Lee DS. Effects of polysaccharides derived from Orostachys japonicus on induction of cell cycle arrest and apoptotic cell death in human colon cancer cells. BMB Rep. 2010;43:750-755. https://doi.org/10.5483/BMBRep.2010.43.11.750
  22. Ryu DS, Lee HS, Lee GS, Lee DS. Effects of the ethylacetate extract of Orostachys japonicus on induction of apoptosis through the p53-mediated signaling pathway in human gastric cancer cells. Biol Pharm Bull. 2012;35:660-665. https://doi.org/10.1248/bpb.35.660
  23. Kim SG, Choi J, Park HJ, Lee SM, Jung HJ. Anti-hyperlipidermic Effects of the flavonoid-rich fraction from the methanol extract of Orostachy japonicus in rats. Kor J Pharmacogn. 2009;40:51-58.
  24. Jeong JH, Ryu DS, Suk DH, Lee DS. Anti-inflammatory effects of ethanol extract from Orostachys japonicus on modulation of signal pathways in LPS-stimulated RAW 264.7 cells. BMB Rep. 2011;44:399-404. https://doi.org/10.5483/BMBRep.2011.44.6.399
  25. Liu F, Kim JK, Li Y, Liu XQ, Li J, Chen X. An extract of Lagerstroemia speciosa L. has insulin-like glucose uptake-stimulatory and adipocyte differentiation-inhibitory activities in 3T3-L1 cells. J Nutrition. 2001;131:2242-2247. https://doi.org/10.1093/jn/131.9.2242
  26. Adorni MP, Cipollari E, Favari E, Zanotti I, Zimmetti F, Corsini A, et al. Inhibitory effect of PCSK9 on ABCA1 protein expression and cholesterol efflux in macrophages. Atherosclerosis. 2017;256:1-6. https://doi.org/10.1016/j.atherosclerosis.2016.11.019
  27. McAlpine CS, Huang A, Emdin A, Banko NS, Beriault DR, Shi Y, et al. Deletion of myeloid $GSK3{\alpha}$ attenuates atherosclerosis and promotes an M2 macrophage phenotype. Arterioscler Thromb Vasc Biol. 2015;35:1113-1122. https://doi.org/10.1161/ATVBAHA.115.305438
  28. Matveev S, van der Westhuyzen DR, Smart EJ. Co-expression of scavenger receptor-BI and caveolin-1 is associated with enhanced selective cholesteryl ester uptake in THP-1 macrophages. J Lipid Res. 1999;40:1647-1654. https://doi.org/10.1016/S0022-2275(20)33410-6
  29. Arakawa R, Abe-Dohmae S, Asai M, Ito JI, Yokoyama S. Involvement of caveolin-1 in cholesterol enrichment of high density lipoprotein during its assembly by apolipoprotein and THP-1 cells. J Lipid Res. 2000;41:1952-1962. https://doi.org/10.1016/S0022-2275(20)32356-7
  30. Chinetti G, Lestavel S, Bocher V. Remaley AT, Neve B, Torra IP, et al. PPAR-alpha and PPAR-gamma activators induce cholesterol removal from human macropharge foam cells through stimulation of the ABCA1 pathway. Nat Med. 2001;7:53-58. https://doi.org/10.1038/83348
  31. Parks JS, Chung SK, Shelnessa GS. Hepatic ABC transporters and triglyceride metabolism. Curr Opin Lipidol. 2012;23:196-200. https://doi.org/10.1097%2FMOL.0b013e328352dd1a https://doi.org/10.1097/MOL.0b013e328352dd1a
  32. Farquhar JW. Sokolow, M. Response of serum lipids and lipoproteins of man to beta-sitosterol and safflower oil. Circulation. 1958;17:890-899. https://doi.org/10.1161/01.cir.17.5.890