Journal of Veterinary Science

The Korean Society of Veterinary Science (대한수의학회)
권호 표지
DOI QR코드

DOI QR Code

Anti-adipogenic effect of the flavonoids through the activation of AMPK in palmitate (PA)-treated HepG2 cells

  • Rajan, Priyanka (College of Veterinary Medicine, Jeju National University) ;
  • Natraj, Premkumar (College of Veterinary Medicine, Jeju National University) ;
  • Ranaweera, Sachithra S. (College of Veterinary Medicine, Jeju National University) ;
  • Dayarathne, Lakshi A. (College of Veterinary Medicine, Jeju National University) ;
  • Lee, Young Jae (College of Veterinary Medicine, Jeju National University) ;
  • Han, Chang-Hoon (College of Veterinary Medicine, Jeju National University)
  • Received : 2021.09.28
  • Accepted : 2021.11.03
  • Published : 2022.01.31
Download PDF
⟨ Previous Next ⟩

Abstract

Background: Flavonoids are natural polyphenols found widely in citrus fruit and peel that possess anti-adipogenic effects. On the other hand, the detailed mechanisms for the antiadipogenic effects of flavonoids are unclear. Objectives: The present study observed the anti-adipogenic effects of five major citrus flavonoids, including hesperidin (HES), narirutin (NAR), nobiletin (NOB), sinensetin (SIN), and tangeretin (TAN), on AMP-activated protein kinase (AMPK) activation in palmitate (PA)-treated HepG2 cells. Methods: The intracellular lipid accumulation and triglyceride (TG) contents were quantified by Oil-red O staining and TG assay, respectively. The glucose uptake was assessed using 2-[N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose (2-NBDG) assay. The levels of AMPK, acetyl-CoA carboxylase (ACC), and glycogen synthase kinase 3 beta (GSK3β) phosphorylation, and levels of sterol regulatory element-binding protein 2 (SREBP-2) and 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) expression were analyzed by Western blot analysis. The potential interaction between the flavonoids and the γ-subunit of AMPK was investigated by molecular docking analysis. Results: The flavonoid treatment reduced both intracellular lipid accumulation and TG content in PA-treated HepG2 cells significantly. In addition, the flavonoids showed increased 2-NBDG uptake in an insulin-independent manner in PA-treated HepG2 cells. The flavonoids increased the AMPK, ACC, and GSK3β phosphorylation levels and decreased the SREBP-2 and HMGCR expression levels in PA-treated HepG2 cells. Molecular docking analysis showed that the flavonoids bind to the CBS domains in the regulatory γ-subunit of AMPK with high binding affinities and could serve as potential AMPK activators. Conclusion: The overall results suggest that the anti-adipogenic effect of flavonoids on PA-treated HepG2 cells results from the activation of AMPK by flavonoids.

Keywords

Acknowledgement

This work was supported by Korean Institute of Planning and Evaluation for Technology in Food, Agriculture and Forestry (IPET) through Innovational Food Technology Development Program, funded by Ministry of Agriculture, Food and Rural Affairs (MAFRA) (grant No. 11901303).

References

  1. Ahmad B, Serpell CJ, Fong IL, Wong EH. Molecular mechanisms of adipogenesis: the anti-adipogenic role of AMP-activated protein kinase. Front Mol Biosci. 2020;7:76. https://doi.org/10.3389/fmolb.2020.00076
  2. Jimenez-Sanchez C, Olivares-Vicente M, Rodriguez-Perez C, Herranz-Lopez M, Lozano-Sanchez J, SeguraCarretero A, et al. AMPK modulatory activity of olive-tree leaves phenolic compounds: bioassay-guided isolation on adipocyte model and in silico approach. PLoS One. 2017;12(3):e0173074. https://doi.org/10.1371/journal.pone.0173074
  3. Wang Q, Liu S, Zhai A, Zhang B, Tian G. AMPK-mediated regulation of lipid metabolism by phosphorylation. Biol Pharm Bull. 2018;41(7):985-993. https://doi.org/10.1248/bpb.b17-00724
  4. Zhu D, Zhang N, Zhou X, Zhang M, Liu Z, Liu X. Cichoric acid regulates the hepatic glucose homeostasis via AMPK pathway and activates the antioxidant response in high glucose-induced hepatocyte injury. RSC Advances. 2017;7(3):1363-1375. https://doi.org/10.1039/C6RA25901D
  5. Yuan HD, Kim DY, Quan HY, Kim SJ, Jung MS, Chung SH. Ginsenoside Rg2 induces orphan nuclear receptor SHP gene expression and inactivates GSK3β via AMP-activated protein kinase to inhibit hepatic glucose production in HepG2 cells. Chem Biol Interact. 2012;195(1):35-42. https://doi.org/10.1016/j.cbi.2011.10.006
  6. Xin FJ, Wang J, Zhao RQ, Wang ZX, Wu JW. Coordinated regulation of AMPK activity by multiple elements in the α-subunit. Cell Res. 2013;23(10):1237-1240. https://doi.org/10.1038/cr.2013.121
  7. Ross FA, MacKintosh C, Hardie DG. AMP-activated protein kinase: a cellular energy sensor that comes in 12 flavours. FEBS J. 2016;283(16):2987-3001. https://doi.org/10.1111/febs.13698
  8. Yong Y, Shin SY, Jung Y, Jung H, Ahn S, Chong Y, et al. Flavonoids activating adenosine monophosphate-activated protein kinase. J Korean Soc Appl Biol Chem. 2015;58(1):13-19. https://doi.org/10.1007/s13765-015-0003-4
  9. Yang Y, Wu Y, Zou J, Wang YH, Xu MX, Huang W, et al. Naringenin attenuates non-alcoholic fatty liver disease by enhancing energy expenditure and regulating autophagy via AMPK. Front Pharmacol. 2021;12:687095. https://doi.org/10.3389/fphar.2021.687095
  10. Kim DS, Lim SB. Extraction of flavanones from immature Citrus unshiu pomace: process optimization and antioxidant evaluation. Sci Rep. 2020;10(1):19950. https://doi.org/10.1038/s41598-020-76965-8
  11. Qi G, Guo R, Tian H, Li L, Liu H, Mi Y, et al. Nobiletin protects against insulin resistance and disorders of lipid metabolism by reprogramming of circadian clock in hepatocytes. Biochim Biophys Acta Mol Cell Biol Lipids. 2018;1863(6):549-562.
  12. Su D, Liu H, Qi X, Dong L, Zhang R, Zhang J. Citrus peel flavonoids improve lipid metabolism by inhibiting miR-33 and miR-122 expression in HepG2 cells. Biosci Biotechnol Biochem. 2019;83(9):1747-1755. https://doi.org/10.1080/09168451.2019.1608807
  13. Xiong H, Wang J, Ran Q, Lou G, Peng C, Gan Q, et al. Hesperidin: a therapeutic agent for obesity. Drug Des Devel Ther. 2019;13:3855-3866. https://doi.org/10.2147/DDDT.S227499
  14. Zhao NQ, Li XY, Wang L, Feng ZL, Li XF, Wen YF, et al. Palmitate induces fat accumulation by activating C/EBPβ-mediated G0S2 expression in HepG2 cells. World J Gastroenterol. 2017;23(43):7705-7715. https://doi.org/10.3748/wjg.v23.i43.7705
  15. Malik SA, Acharya JD, Mehendale NK, Kamat SS, Ghaskadbi SS. Pterostilbene reverses palmitic acid mediated insulin resistance in HepG2 cells by reducing oxidative stress and triglyceride accumulation. Free Radic Res. 2019;53(7):815-827. https://doi.org/10.1080/10715762.2019.1635252
  16. Liu JY, Zhang YC, Song LN, Zhang L, Yang FY, Zhu XR, et al. Nifuroxazide ameliorates lipid and glucose metabolism in palmitate-induced HepG2 cells. RSC Advances. 2019;9(67):39394-39404. https://doi.org/10.1039/c9ra06527j
  17. Kramer B, Rarey M, Lengauer T. Evaluation of the FLEXX incremental construction algorithm for proteinligand docking. Proteins. 1999;37(2):228-241. https://doi.org/10.1002/(SICI)1097-0134(19991101)37:2<228::AID-PROT8>3.0.CO;2-8
  18. Kim MS, Hur HJ, Kwon DY, Hwang JT. Tangeretin stimulates glucose uptake via regulation of AMPK signaling pathways in C2C12 myotubes and improves glucose tolerance in high-fat diet-induced obese mice. Mol Cell Endocrinol. 2012;358(1):127-134. https://doi.org/10.1016/j.mce.2012.03.013
  19. Kahn BB, Alquier T, Carling D, Hardie DG. AMP-activated protein kinase: ancient energy gauge provides clues to modern understanding of metabolism. Cell Metab. 2005;1(1):15-25. https://doi.org/10.1016/j.cmet.2004.12.003
  20. Carling D. The AMP-activated protein kinase cascade--a unifying system for energy control. Trends Biochem Sci. 2004;29(1):18-24. https://doi.org/10.1016/j.tibs.2003.11.005
  21. Chen L, Duan Y, Wei H, Ning H, Bi C, Zhao Y, et al. Acetyl-CoA carboxylase (ACC) as a therapeutic target for metabolic syndrome and recent developments in ACC1/2 inhibitors. Expert Opin Investig Drugs. 2019;28(10):917-930. https://doi.org/10.1080/13543784.2019.1657825
  22. Foretz M, Viollet B. Activation of AMPK for a break in hepatic lipid accumulation and circulating cholesterol. EBioMedicine. 2018;31:15-16. https://doi.org/10.1016/j.ebiom.2018.05.009
  23. Yuk T, Kim Y, Yang J, Sung J, Jeong HS, Lee J. Nobiletin inhibits hepatic lipogenesis via activation of AMPactivated protein kinase. Evid Based Complement Alternat Med. 2018;2018:7420265.
  24. Luo J, Yang H, Song BL. Mechanisms and regulation of cholesterol homeostasis. Nat Rev Mol Cell Biol. 2020;21(4):225-245. https://doi.org/10.1038/s41580-019-0190-7
  25. Sakakura Y, Shimano H, Sone H, Takahashi A, Inoue N, Toyoshima H, et al. Sterol regulatory elementbinding proteins induce an entire pathway of cholesterol synthesis. Biochem Biophys Res Commun. 2001;286(1):176-183. https://doi.org/10.1006/bbrc.2001.5375
  26. Lu J, Meng Z, Cheng B, Liu M, Tao S, Guan S. Apigenin reduces the excessive accumulation of lipids induced by palmitic acid via the AMPK signaling pathway in HepG2 cells. Exp Ther Med. 2019;18(4):2965-2971.
  27. Ha T, Trung TN, Hien TT, Dao TT, Yim N, Ngoc TM, et al. Selected compounds derived from Moutan Cortex stimulated glucose uptake and glycogen synthesis via AMPK activation in human HepG2 cells. J Ethnopharmacol. 2010;131(2):417-424. https://doi.org/10.1016/j.jep.2010.07.010
  28. Zhang J, Sun C, Yan Y, Chen Q, Luo F, Zhu X, et al. Purification of naringin and neohesperidin from Huyou (Citrus changshanensis) fruit and their effects on glucose consumption in human HepG2 cells. Food Chem. 2012;135(3):1471-1478. https://doi.org/10.1016/j.foodchem.2012.06.004
  29. Xiao B, Heath R, Saiu P, Leiper FC, Leone P, Jing C, et al. Structural basis for AMP binding to mammalian AMP-activated protein kinase. Nature. 2007;449(7161):496-500. https://doi.org/10.1038/nature06161
  30. Carling D, Mayer FV, Sanders MJ, Gamblin SJ. AMP-activated protein kinase: nature's energy sensor. Nat Chem Biol. 2011;7(8):512-518. https://doi.org/10.1038/nchembio.610
  31. Jeon SM. Regulation and function of AMPK in physiology and diseases. Exp Mol Med. 2016;48(7):e245. https://doi.org/10.1038/emm.2016.81