Journal of Biomedical Engineering Research (대한의용생체공학회:의공학회지)

The Korean Society of Medical and Biological Engineering (대한의용생체공학회)
권호 표지
DOI QR코드

DOI QR Code

Effects of Micro-current Stimulation on lipid metabolism in Oleic Acid-Induced Non-Alcoholic Fatty Liver disease in FL83B cells

올레산으로 유도된 비알코올성 지방간 세포 모델에서의 미세전류 자극의 지질 대사 조절 효능 평가

  • Lee, Hana (Department of Biomedical Engineering, Yonsei University) ;
  • Lee, Minjoo (Department of Biomedical Engineering, Yonsei University) ;
  • Kim, Han Sung (Department of Biomedical Engineering, Yonsei University)
  • Received : 2022.01.18
  • Accepted : 2022.01.27
  • Published : 2022.02.28
Download PDF
⟨ Previous Next ⟩

Abstract

Non-alcoholic fatty liver disease(NAFLD) is excessive hepatic lipid accumulation mainly caused by obesity. This study aimed to evaluate whether micro-current stimulation(MCS) could modulate lipid metabolism regarding the Sirt1/AMPK pathway, fatty acid β-oxidation pathway, and lipolysis and lipogenesis-related factors in FL83B cells. For the NAFLD cell model, FL83B cells were treated with oleic acid for lipid accumulation. MCS were stimulated for 1 hr and used frequency 10 Hz, duty cycle 50%, and biphasic rectangular current pulse. The intensity of MCS was divided into 50, 100, 200, and 400 ㎂. Through the results of Oil red O staining, it was confirmed that MCSs with the intensity of 200 ㎂ and 400 ㎂ significantly reduced the degree of lipid droplet formation. Thus, these MCS intensities were applied to western blot analysis. Western blot analysis was performed to analyze the effects of MCS on lipid metabolism. MCS with the intensity of 400 ㎂ showed that significantly activated the Sirt1/AMPK pathway, a key pathway for regulating lipid metabolism in hepatocytes, and fatty acid β-oxidation-related transcription factors. Moreover, it activated the lipolysis pathway and suppressed lipogenesis-related transcription factors such as SREBP-1c, FAS, and PPARγ. In the case of MCS with the intensity of 200 ㎂, only PGC1α and SREBP-1c showed significant differences compared to cells treated only with oleic acid. Taken together, these results suggested that MCS with the intensity of 400 ㎂ could alleviate hepatic lipid accumulation by modulating lipid metabolism in hepatocytes.

Keywords

Acknowledgement

본 연구는 정부(과학기술정보통신부)의 재원으로 한국연구재단의 지원을 받아 수행된 연구임(No. 2021R1A2C2093828).

References

  1. Gui Y, Pan Q, Chen X, Xu S, Luo X, Chen L. The association between obesity related adipokines and risk of breast cancer: a meta-analysis. Oncotarget 2017;8(43):75389. https://doi.org/10.18632/oncotarget.17853
  2. Yoo KD, Jun DW. Nonalcoholic Fatty Liver Disease: Lifestyle Modification. Korean Journal of Medicine 2014;86(4):416-424. https://doi.org/10.3904/kjm.2014.86.4.416
  3. Lin HTV, Tsou YC, Chen YT, Lu WJ, Hwang PA. Effects of low-molecular-weight fucoidan and high stability fucoxanthin on glucose homeostasis, lipid metabolism, and liver function in a mouse model of type II diabetes. Marine drugs 2017;15(4):113. https://doi.org/10.3390/md15040113
  4. Kitade H, Chen G, Ni Y, Ota T. Nonalcoholic fatty liver disease and insulin resistance: new insights and potential new treatments. Nutrients 2017;9(4):387. https://doi.org/10.3390/nu9040387
  5. He XX, Wu XL, Chen RP, Chen C, Liu XG, Wu BJ, Huang ZM. Effectiveness of omega-3 polyunsaturated fatty acids in non-alcoholic fatty liver disease: a meta-analysis of randomized controlled trials. PLoS One 2016;11(10):e0162368. https://doi.org/10.1371/journal.pone.0162368
  6. Kim CS, Cho EH, Choe SY, Kang MS, Yu R. Inhibitory effect of isorhamnetin on lipid accumulation in free fatty acid-induced steatotic hepatocytes through the PPARα pathway. Journal of the Korean Society of Food Science and Nutrition 2018;47(7):703-709. https://doi.org/10.3746/jkfn.2018.47.7.703
  7. Raghow R, Yellaturu C, Deng X, Park EA, Elam MB. SREBPs: the crossroads of physiological and pathological lipid homeostasis. Trends in endocrinology & metabolism 2008;19(2):65-73. https://doi.org/10.1016/j.tem.2007.10.009
  8. Yeh YT, Cho YY, Hsieh SC, Chiang AN. Chinese olive extract ameliorates hepatic lipid accumulation in vitro and in vivo by regulating lipid metabolism. Scientific reports 2018;8(1):1-13.
  9. Browning JD, Horton JD. Molecular mediators of hepatic steatosis and liver injury. The Journal of clinical investigation 2004;114(2):147-152. https://doi.org/10.1172/JCI200422422
  10. Tian J, Goldstein JL, Brown MS. Insulin induction of SREBP-1c in rodent liver requires LXRα-C/EBPβ complex. Proceedings of the National Academy of Sciences 2016;113(29):8182-8187. https://doi.org/10.1073/pnas.1608987113
  11. Wang Y, Viscarra J, Kim SJ, Sul HS. Transcriptional regulation of hepatic lipogenesis. Nature reviews Molecular cell biology 2015;16(11):678-689. https://doi.org/10.1038/nrm4074
  12. Long YC, Zierath JR. AMP-activated protein kinase signaling in metabolic regulation. The Journal of clinical investigation 2006;116(7):1776-1783. https://doi.org/10.1172/JCI29044
  13. Foretz M, Viollet B. Regulation of hepatic metabolism by AMPK. Journal of Hepatology 2011;54(4):827-9. https://doi.org/10.1016/j.jhep.2010.09.014
  14. Li YU, Xu S, Mihaylova MM, Zheng B, Hou X, Jiang B, Zang M. AMPK phosphorylates and inhibits SREBP activity to attenuate hepatic steatosis and atherosclerosis in diet-induced insulin-resistant mice. Cell metabolism 2011;13(4):376-388. https://doi.org/10.1016/j.cmet.2011.03.009
  15. Staels B, Fruchart JC. Therapeutic roles of peroxisome proliferator-activated receptor agonists. Diabetes 2005;54(8):2460-2470. https://doi.org/10.2337/diabetes.54.8.2460
  16. Szkudelski T, Szkudelska K. Effects of AMPK activation on lipolysis in primary rat adipocytes: studies at different glucose concentrations. Archives of physiology and biochemistry 2017;123(1):43-49. https://doi.org/10.1080/13813455.2016.1227853
  17. Liou CJ, Wei CH, Chen YL, Cheng CY, Wang CL, Huang WC. Fisetin protects against hepatic steatosis through regulation of the Sirt1/AMPK and fatty acid β-oxidation signaling pathway in high-fat diet-induced obese mice. Cellular physiology and biochemistry 2018;49(5):1870-1884. https://doi.org/10.1159/000493650
  18. Cho MS, Park RJ, Park SH, Cho YH, Cheng GA. The effect of microcurrent-inducing shoes on fatigue and pain in middle-aged people with plantar fascitis. Journal of Physical Therapy Science 2007;19(2):165-170. https://doi.org/10.1589/jpts.19.165
  19. Oh HJ, Kim JY, Park RJ. The effects of microcurrent stimulation on recovery of function and pain in chronic low back pain. Journal of the Korean Society of Physical Medicine 2008;3(1):47-56.
  20. Hwang D, Lee H, Lee M, Cho S, Kim HS. The Micro-Current Stimulation Inhibits Adipogenesis by Activating Wnt/ β-Catenin Signaling. Journal of Biomedical Engineering Research 2020;41(6):235-246. https://doi.org/10.9718/JBER.2020.41.6.235
  21. 김한성 외, 세포 전기자극 시스템 및 방법, 대한민국 특허청 등록특허공보 등록번호 10-2101372, 2020.
  22. Reccia I, Kumar J, Akladios C, Virdis F, Pai M, Habib N, Spalding D. Non-alcoholic fatty liver disease: a sign of systemic disease. Metabolism 2017;72:94-108. https://doi.org/10.1016/j.metabol.2017.04.011
  23. Hazlehurst JM, Woods C, Marjot T, Cobbold JF, Tomlinson JW. Non-alcoholic fatty liver disease and diabetes. Metabolism 2016;65(8):1096-1108. https://doi.org/10.1016/j.metabol.2016.01.001
  24. Heindel JJ, Newbold R, Schug TT. Endocrine disruptors and obesity. Nature Reviews Endocrinology 2015;11(11):653-661. https://doi.org/10.1038/nrendo.2015.163
  25. Yoo A, Narayan VP, Hong EY, Whang WK, Park T. Scopolin ameliorates high-fat diet induced hepatic steatosis in mice: potential involvement of SIRT1-mediated signaling cascades in the liver. Scientific reports 2017;7(1):1-11. https://doi.org/10.1038/s41598-016-0028-x
  26. Herbrechter R, Ziemba PM, Hoffmann KM, Hatt H, Werner M, Gisselmann, G. Identification of Glycyrrhiza as the rikkunshito constituent with the highest antagonistic potential on heterologously expressed 5-HT3A receptors due to the action of flavonoids. Frontiers in pharmacology 2015;6:130.
  27. Wang Z, Pini M, Yao T, Zhou Z, Sun C, Fantuzzi G, Song Z. Homocysteine suppresses lipolysis in adipocytes by activating the AMPK pathway. American journal of physiology-endocrinology and metabolism 2011;301(4):E703-E712. https://doi.org/10.1152/ajpendo.00050.2011
  28. Chen H, Liu X, Chen H, Cao J, Zhang L, Hu X, Wang J. Role of SIRT1 and AMPK in mesenchymal stem cells differentiation. Ageing research reviews 2014;13:55-64. https://doi.org/10.1016/j.arr.2013.12.002
  29. Lim CT, Kola B, Korbonits M. AMPK as a mediator of hormonal signalling. Journal of molecular endocrinology 2010;44(2):87.
  30. Peng IC, Chen Z, Sun W, Li YS, Marin TL, Hsu PH, Shyy JY. Glucagon regulates ACC activity in adipocytes through the CAMKKβ/AMPK pathway. American Journal of Physiology-Endocrinology and Metabolism 2012;302(12):E1560-E1568. https://doi.org/10.1152/ajpendo.00504.2011
  31. Huang WC, Chang WT, Wu SJ, Xu PY, Ting NC, Liou CJ. Phloretin and phlorizin promote lipolysis and inhibit inflammation in mouse 3T3-L1 cells and in macrophage-adipocyte co-cultures. Molecular nutrition & food research 2013;57(10):1803-1813. https://doi.org/10.1002/mnfr.201300001
  32. Malandrino MI, Fucho R, Weber M, Calderon-Dominguez M, Mir JF, Valcarcel L, Herrero L. Enhanced fatty acid oxidation in adipocytes and macrophages reduces lipid-induced triglyceride accumulation and inflammation. American Journal of Physiology-Endocrinology and Metabolism 2015;308(9):E756-E769. https://doi.org/10.1152/ajpendo.00362.2014
  33. Houten SM, Violante S, Ventura FV, Wanders RJ. The biochemistry and physiology of mitochondrial fatty acid β-oxidation and its genetic disorders. Annual review of physiology 2016;78:23-44. https://doi.org/10.1146/annurev-physiol-021115-105045
  34. Brunt EM. Nonalcoholic fatty liver disease: pros and cons of histologic systems of evaluation. International journal of molecular sciences 2016;17(1):97. https://doi.org/10.3390/ijms17010097
  35. Choi SS, Diehl AM. Hepatic triglyceride synthesis and nonalcoholic fatty liver disease. Current opinion in lipidology 2008;19(3):295-300. https://doi.org/10.1097/MOL.0b013e3282ff5e55
  36. Cherkaoui-Malki M, Surapureddi S, I El Hajj H, Vamecq J, Andreoletti P. Hepatic steatosis and peroxisomal fatty acid beta-oxidation. Current Drug Metabolism 2012;13(10):1412-1421. https://doi.org/10.2174/138920012803762765
  37. Evans RM, Barish GD, Wang YX. PPARs and the complex journey to obesity. Nature medicine 2004;10(4):355-361. https://doi.org/10.1038/nm1025
  38. Postic C, Girard J. Contribution of de novo fatty acid synthesis to hepatic steatosis and insulin resistance: lessons from genetically engineered mice. The Journal of clinical investigation 2008;118(3):829-838. https://doi.org/10.1172/JCI34275
  39. Finck BN, Kelly DP. PGC-1 coactivators: inducible regulators of energy metabolism in health and disease. The Journal of clinical investigation 2006;116(3):615-622. https://doi.org/10.1172/JCI27794
  40. Day EA, Ford RJ, Steinberg GR. AMPK as a therapeutic target for treating metabolic diseases. Trends in Endocrinology & Metabolism 2017;28(8):545-560. https://doi.org/10.1016/j.tem.2017.05.004
  41. Smith BK, Marcinko K, Desjardins EM, Lally JS, Ford RJ, Steinberg GR. Treatment of nonalcoholic fatty liver disease: role of AMPK. American Journal of Physiology-Endocrinology and Metabolism 2016;311(4):E730-E740. https://doi.org/10.1152/ajpendo.00225.2016
  42. Ducharme NA, Bickel PE. Minireview: lipid droplets in lipogenesis and lipolysis. Endocrinology 2008;149(3):942-949. https://doi.org/10.1210/en.2007-1713
  43. Fruhbeck G, Mendez-Gimenez L, Fernandez-Formoso JA, Fernandez S, Rodriguez A. Regulation of adipocyte lipolysis. Nutrition research reviews 2014;27(1):63-93. https://doi.org/10.1017/S095442241400002X
  44. Malandrino MI, Fucho R, Weber M, Calderon-Dominguez M, Mir JF, Valcarcel L, Herrero L. Enhanced fatty acid oxidation in adipocytes and macrophages reduces lipid-induced triglyceride accumulation and inflammation. American Journal of Physiology-Endocrinology and Metabolism 2015;308(9):E756-E769. https://doi.org/10.1152/ajpendo.00362.2014
  45. Duncan RE, Ahmadian M, Jaworski K, Sarkadi-Nagy E, Sul HS. Regulation of lipolysis in adipocytes. Annu. Rev. Nutr. 2007;27:79-101. https://doi.org/10.1146/annurev.nutr.27.061406.093734
  46. Lampidonis AD, Rogdakis E, Voutsinas GE, Stravopodis DJ. The resurgence of Hormone-Sensitive Lipase (HSL) in mammalian lipolysis. Gene 2011;477(1-2):1-11. https://doi.org/10.1016/j.gene.2011.01.007
  47. Gaidhu MP, Fediuc S, Anthony NM, So M, Mirpourian M, Perry RL, Ceddia RB. Prolonged AICAR-induced AMPkinase activation promotes energy dissipation in white adipocytes: novel mechanisms integrating HSL and ATGL. Journal of lipid research 2009;50(4):704-715. https://doi.org/10.1194/jlr.M800480-JLR200
  48. Li Y, Zhu W, Li J, Liu M, Wei M. Resveratrol suppresses the STAT3 signaling pathway and inhibits proliferation of high glucose-exposed HepG2 cells partly through SIRT1. Oncology reports 2013;30(6):2820-2828. https://doi.org/10.3892/or.2013.2748