한국식품과학회지 (Korean Journal of Food Science and Technology)

한국식품과학회 (Korean Society of Food Science and Technology)
권호 표지
DOI QR코드

DOI QR Code

홍삼 사포닌 분획의 Nrf2 Keap1 신호전달체계 조절을 통한 지방축적 및 활성산소종 억제효과

Red ginseng-derived saponin fraction inhibits lipid accumulation and reactive oxygen species production by activating nuclear factor (erythroid-derived 2)-like 2 (Nrf2)/Kelch-like ECH-associated protein 1 (Keap1) pathway

  • 김채영 (고려대학교 보건과학대 의생명융합과학과) ;
  • 강보빈 (고려대학교 보건과학대 의생명융합과학과) ;
  • 황지수 (고려대학교 보건과학대 의생명융합과학과) ;
  • 최현선 (서울여자대학교 식품공학과)
  • Kim, Chae-Young (Department of Integrated Biomedical and Life Science, Graduate School, Korea University) ;
  • Kang, Bobin (Department of Integrated Biomedical and Life Science, Graduate School, Korea University) ;
  • Hwang, Jisu (Department of Integrated Biomedical and Life Science, Graduate School, Korea University) ;
  • Choi, Hyeon-Son (Department of Food Science and Technology, College of Natural Science, Seoul Women's University)
  • 투고 : 2018.09.18
  • 심사 : 2018.11.28
  • 발행 : 2018.12.31
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

본 연구에서는 홍삼 사포닌 분획(SF)으로부터 진세노사이드의 조성을 분석하고 지방세포의 분화 및 지방축적에 대한 효과를 측정하였다. SF는 지방분화인자인 $PPAR{\gamma}$, $C/EBP{\alpha}$의 단백질 양을 억제함으로써 지방분화 동안 효과적으로 지방축적을 억제하였으며 주로 지방분화 초기시점부터 지방분화 초기인자인 $C/EBP{\beta}$, KLF2의 조절작용을 통해 지방축적을 억제하는 것으로 관찰되었다. SF는 또한 지방분화 동안 생성되는 ROS의 생성을 효과적으로 억제하였는데 이는 SF가 산화방지 시스템인 Nrf2/Keap1 경로를 활성화하기 때문으로 판단되며 특히 Nrf2의 핵 내로의 진입을 활성화 함으로써 Nrf2의 타겟 산화방지 분자들인 HO-1, NQO1의 발현을 촉진하였다. 이는 지방분화 동안 SF의 지방축적 억제 효과가 Nrf2의 활성화와 밀접하게 관련이 있음을 보여준다.

This study aimed to investigate the effects of red ginseng-derived saponin fraction (SF) on lipid accumulation, reactive oxygen species (ROS) production, and nuclear factor (erythroid-derived 2)-like 2 (Nrf2)/Kelch-like ECH-associated protein 1 (Keap1) signaling during adipocyte differentiation. SF effectively inhibited lipid accumulation, with the downregulation of adipogenic factors such as peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) and CCAAT/enhancer-binding protein alpha ($C/EBP{\alpha}$). A high dose of SF decreased the protein levels of $PPAR{\gamma}$ and $C/EBP{\alpha}$ by over 90% compared to the control. SF-mediated downregulation of adipogenic factors was due to the regulation of early adipogenic factors including $C/EBP{\beta}$ and $Kr{\ddot{u}}ppel$-like Factor 2 (KLF2). In addition, SF ($200{\mu}g/mg$) decreased intracellular ROS generation by 40% during adipocyte differentiation. However, the SF significantly upregulated Nrf2 and its target proteins, hemoxygenase-1 (HO-1) and NADPH dehydrogenase quinone 1 (NQO1). Furthermore, SF ($200{\mu}g/mg$) promoted the nuclear translocation of Nrf2. The SF-mediated reduction of lipid accumulation was associated with the regulation of the Nrf2/Keap1 pathway.

키워드

SPGHB5_2018_v50n6_688_f0001.png 이미지

Fig. 1. Effect of saponin fraction (SF) on viability of 3T3-L1 cells.

SPGHB5_2018_v50n6_688_f0002.png 이미지

Fig. 2. Effect of saponin fraction (SF) on lipid accumulation during adipocyte differentiation.

SPGHB5_2018_v50n6_688_f0003.png 이미지

Fig. 3. Effect of saponin fraction (SF) on adipogenic stages.

SPGHB5_2018_v50n6_688_f0004.png 이미지

Fig. 5. Effect of saponin fraction (SF) on the production of ROS in 3T3-L1 cells.

SPGHB5_2018_v50n6_688_f0005.png 이미지

Fig. 6. Effect of saponin fraction (SF) on the protein expression level of Nrf2, Keap1, HO-1 and NQO1, and Nrf2 nuclear translocation in 3T3-L1 cells.

SPGHB5_2018_v50n6_688_f0006.png 이미지

Fig. 7. Effect of saponin fraction (SF) on the protein expression level of AKT, P-AKT in 3T3-L1 cells.

SPGHB5_2018_v50n6_688_f0007.png 이미지

Fig. 4. Effect of saponin fraction (SF) on early adipogenic factors.

Table 1. Ginsenoside composition from saponin fraction

SPGHB5_2018_v50n6_688_t0001.png 이미지

Table 2. Radical scavenging activity of saponin fraction

SPGHB5_2018_v50n6_688_t0002.png 이미지

참고문헌

  1. Attele AS, Wu JA, Yuan CS. Ginseng pharmacology: multiple constituents and multiple actions. Biochem. Pharmacol. 58: 1685-1693 (1995)
  2. Chang H. Effect of processing methods on the saponin contents of Panax ginseng leaf-tea. J. Food Sci. Nut.16: 46-53 (2003)
  3. Chang YS, Chang YH, Sung JH. The effect of ginseng and caffeine products on the antioxidative activities of mouse kidney. J. Ginseng Res. 30: 15-21 (2006) https://doi.org/10.5142/JGR.2006.30.1.015
  4. Chen HH, Chen YT, Huang YW, Tsai HJ, Kuo CC. 4-Ketopinoresinol, a novel naturally occurring ARE activator, induces the Nrf2/HO-1 axis and protects against oxidative stress-induced cell injury via activation of PI3K/AKT signaling. Free Radic. Biol. Med. 52: 1054-1066 (2012) https://doi.org/10.1016/j.freeradbiomed.2011.12.012
  5. Choi HS, Jeon HJ, Lee OH, Lee BY. Indole-3-carbinol, a vegetable phytochemical, inhibits adipogenesis by regulating cell cycle and $AMPK{\alpha}$ signaling. Biochimie. 104: 127-136 (2014) https://doi.org/10.1016/j.biochi.2014.06.010
  6. Choi KM, Lee YS, Sin DM, Lee S, Lee MK, Lee YM, Hong JT, Yun YP, Yoo HS. Sulforaphane inhibits mitotic clonal expansion during adipogenesis through cell cycle arrest. Obesity. 20: 1365-1371 (2012) https://doi.org/10.1038/oby.2011.388
  7. Furukawa S, Fujita T, Shimabukuro M, Iwaki M, Yamada Y, Nakajima Y, Nakayama O, Makishima M, Matsuda M, Shimomura I. Increased oxidative stress in obesity and its impact on metabolic syndrome. J. Clin. Invest. 114: 1752-1761 (2017)
  8. Gaikwad A, Long DJ, Stringer JL, Jaiswal AK. In Vivo Role of NAD (P) H: quinone oxidoreductase1 (NQO1) in the regulation of intracellular redox state and accumulation of abdominal adipose tissue. J. Biol. Chem. 276: 22559-22564 (2001) https://doi.org/10.1074/jbc.M101053200
  9. Halliwell B, Gutteridge JM, Cross CE. Free radicals, antioxidants, and human disease: where are we now?. J. Lab.Clin. Med. 119: 598-620 (1992)
  10. He HJ, Wang GY, Gao Y, Ling WH, Yu ZW, Jin TR. Curcumin attenuates Nrf2 signaling defect, oxidative stress in muscle and glucose intolerance in high fat diet-fed mice. World J. Diabetes 3: 94-104 (2012) https://doi.org/10.4239/wjd.v3.i5.94
  11. Kersten S. Mechanisms of nutritional and hormonal regulation of lipogenesis. EMBO Reports. 2: 282-286 (2001) https://doi.org/10.1093/embo-reports/kve071
  12. Keum YS, Park KK, Lee JM, Chun KS, Park JH, Lee SK, Kwon HJ, Surh YJ. Antioxidant and anti-tumor promoting activities of the methanol extract of heat-processed ginseng. Cancer Letters. 150: 41-48 (2000) https://doi.org/10.1016/S0304-3835(99)00369-9
  13. Kim DH, Kwak KH, Lee KJ, Kim SJ, Shin YC, Chun BG, Shin, KH. Effects of Korea red ginseng total saponin on repeated unpredictable stress-induced changes of proliferation of neural progenitor cells and BDNF mRNA expression in adult rat hippocampus. J. Ginseng Res. 28: 94-103 (2004). https://doi.org/10.5142/JGR.2004.28.2.094
  14. Kim JH, Kang S, Jung YN, Choi H-S. Cholecalciferol inhibits lipid accumulation by regulating early adipogenesis in cultured adipocytes and zebrafish. Biochem. Biophys. Res. Commun. 469: 646-653 (2016) https://doi.org/10.1016/j.bbrc.2015.12.049
  15. Kim JY, Park JY, Kang HJ, Kim OY, Lee JH. Beneficial effects of Korean red ginseng on lymphocyte DNA damage, antioxidant enzyme activity, and LDL oxidation in healthy participants: a randomized, double-blind, placebo-controlled trial. Nutr. J.11: 47 (2012) https://doi.org/10.1186/1475-2891-11-47
  16. Konno C, Hikino H. Isolation and hypoglycemic activity of panaxans M, N, O and P, glycans of Panax ginseng roots. Int. J. Crude Drug Res. 25: 53-56 (1987) https://doi.org/10.3109/13880208709060912
  17. Lee H, Lee YJ, Choi H, Ko EH, Kim JW. Reactive oxygen species facilitate adipocyte differentiation by accelerating mitotic clonal expansion. J. Biol. Chem. 284: 10601-10609 (2009) https://doi.org/10.1074/jbc.M808742200
  18. Lee JW, Do JH. Antioxidative activity of ethanol extraction fraction from the Korean red tail ginseng. Kor. J. Food Sci. Technol. 33: 497-500 (2001).
  19. Lee M, Kim I, Kim C, Kim Y. Effects of ginsenoside Rg3 on adipocyte fatty acid binding protein mRNA expression and glycerol-3-phosphate dehydrogenase activity during adipocytes differentiation. Kor. J. Lipidol. 21: 67-75 (2011)
  20. Liu H, Wang J, Liu M, Zhao H, Yaqoob S, Zheng M, Cai D, Liu J. Antiobesity effects of ginsenoside Rg1 on 3T3-L1 preadipocytes and high fat diet-induced obese mice mediated by AMPK. Nutrients 10: 830-844 (2018) https://doi.org/10.3390/nu10070830
  21. Lowe CE, O'Rahilly S, Rochford JJ. Adipogenesis at a glance. J. Cell Sci. 124: 2681-2686 (2011) https://doi.org/10.1242/jcs.079699
  22. Nam KY. The comparative understanding between red ginseng and white ginsengs, processed ginsengs (Panax ginseng CA Meyer). J. Ginseng Res. 29: 1-18 (2005) https://doi.org/10.5142/JGR.2005.29.1.001
  23. Oakley FD, Abbott D, Li Q, Engelhardt JF. Signaling components of redox active endosomes: the redoxosomes. Antioxid. Redox Signal. 11: 1313-1333 (2009) https://doi.org/10.1089/ars.2008.2363
  24. Panchal SK, Poudyal H, Brown L. Quercetin ameliorates cardiovascular, hepatic, and metabolic changes in diet-induced metabolic syndrome in rats. J. Nutr. 142: 1026-1032 (2012) https://doi.org/10.3945/jn.111.157263
  25. Pi J, Leung L, Xue P, Wang W, Hou Y, Liu D,Yehuda-Shnaidman E, Lee, C, Lau J, Kurtz T W, Chan JY. Deficiency in the nuclear factor E2-related factor 2 transcription factor results in impaired adipogenesis and protects against diet-induced obesity. J. Biol. Chem. 285: 9292-9300 (2010) https://doi.org/10.1074/jbc.M109.093955
  26. Rosen ED, Walkey CJ, Puigserver P, Spiegelman BM. Transcriptional regulation of adipogenesis. Genes Dev. 14: 1293-1307 (2000)
  27. Shin S, Wakabayashi N, Misra V, Biswal S, Lee GH, Agoston ES, Yamamoto M, Kensler TW. NRF2 modulates aryl hydrocarbon receptor signaling: influence on adipogenesis. Mol. Cell. Biol. 27: 7188-7197 (2007) https://doi.org/10.1128/MCB.00915-07
  28. Spiegelman BM, Choy L, Hotamisligil GS, Graves RA, Tontonoz P. Regulation of adipocyte gene expression in differentiation and syndromes of obesity/diabetes. J. Biol. Chem. 268: 6823-6826 (1993)
  29. Suh HJ, Cho SY, Kim EY, Choi HS. Blockade of lipid accumulation by silibinin in adipocytes and zebrafish. Chem. Biol. Interact. 227: 53-62 (2015) https://doi.org/10.1016/j.cbi.2014.12.027
  30. Taguchi K, Motohashi H, Yamamoto M. Molecular mechanisms of the Keap1-Nrf2 pathway in stress response and cancer evolution. Genes Cells. 16: 123-140 (2011) https://doi.org/10.1111/j.1365-2443.2010.01473.x
  31. Tang W, Yan J, Wang T, Xia X, Zhuang X, Hong K, Li R, Liu P, Jiang H, Qiao J. Up-regulation of heme oxygenase-1 expression modulates reactive oxygen species level during the cryopreservation of human seminiferous tubules. Fertil. Steril. 102: 974-980 (2014) https://doi.org/10.1016/j.fertnstert.2014.07.736
  32. Wang L, Chen Y, Sternberg P, Cai J. Essential roles of the PI3 kinase/Akt pathway in regulating Nrf2-dependent antioxidant functions in the RPE. Invest. Ophthalmol. Vis. Sci. 49: 1671-1678 (2008) https://doi.org/10.1167/iovs.07-1099
  33. Wu Y, Huang XF, Bell C, Yu Y. Ginsenoside Rb1 improves leptin sensitivity in the prefrontal cortex in obese mice. CNS Neurosci. Ther. 24: 98-107 (2018) https://doi.org/10.1111/cns.12776
  34. Yoon S, Joo C. Study on the preventive effect of ginsenosides against hypercholesterolemia and its mechanism. Kor. J. Ginseng Sci. 17: 1-12 (1993)
  35. Yuan Q, Jiang YW, Ma TT, Fang QH, Pan L. Attenuating effect of Ginsenoside Rb1 on LPS-induced lung injury in rats. J. Inflammation. 11: 40 (2014) https://doi.org/10.1186/s12950-014-0040-5
  36. Zhang L, Zhang L, Wang X, Si H. Anti-adipogenic effects and mechanisms of ginsenoside Rg3 in pre-adipocytes and obese mice. Front. Pharmacol. 8: 113 (2017)
  37. Zhang M, An C, Gao Y, Leak RK, Chen J, Zhang F. Emerging roles of Nrf2 and phase II antioxidant enzymes in neuroprotection. Prog. Neurobiol.100: 30-47 (2013) https://doi.org/10.1016/j.pneurobio.2012.09.003
  38. Zhong Y, Liu T, Lai W, Tan Y, Tian D, Guo Z. Heme oxygenase-1-mediated reactive oxygen species reduction is involved in the inhibitory effect of curcumin on lipopolysaccharide-induced monocyte chemoattractant protein-1 production in RAW264.7 macrophages. Molecular Med. Reports. 7: 242-246 (2013) https://doi.org/10.3892/mmr.2012.1138