DOI QR코드

DOI QR Code

Curcumin represses lipid accumulation through inhibiting ERK1/2-PPAR-γ signaling pathway and triggering apoptosis in porcine subcutaneous preadipocytes

  • Pan, Shifeng (College of Veterinary Medicine, Yangzhou University) ;
  • Chen, Yongfang (College of Veterinary Medicine, Yangzhou University) ;
  • Zhang, Lin (College of Veterinary Medicine, Yangzhou University) ;
  • Liu, Zhuang (College of Veterinary Medicine, Yangzhou University) ;
  • Xu, Xingyu (College of Veterinary Medicine, Yangzhou University) ;
  • Xing, Hua (College of Veterinary Medicine, Yangzhou University)
  • 투고 : 2021.08.17
  • 심사 : 2021.10.19
  • 발행 : 2022.05.01

초록

Objective: Excessive lipid accumulation in adipocytes results in prevalence of obesity and metabolic syndrome. Curcumin (CUR), a naturally phenolic active ingredient, has been shown to have lipid-lowering effects. However, its underlying mechanisms have remained largely unknown. Therefore, the study aims to determine the effect of CUR on cellular lipid accumulation in porcine subcutaneous preadipocytes (PSPA) and to clarify novel mechanisms. Methods: The PSPA were cultured and treated with or without CUR. Both cell counting Kit-8 and lactate dehydrogenase release assays were used to examine cytotoxicity. Intracellular lipid contents were measured by oil-red-o staining extraction and triglyceride quantification. Apoptosis was determined by flow cytometry and the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-nick end labelling assay. Adipogenic and apoptosis genes were analyzed by quantitative polymerase chain reaction and Western blot. Results: The CUR dose-dependently reduced the proliferation and lipid accumulation of PSPA. Noncytotoxic doses of CUR (10 to 20 μM) significantly inhibited extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation and expression of adipogenic genes peroxisome proliferation-activity receptor-γ (PPAR-γ), CCAAT/enhancer binding protein-α, sterol regulatory element-binding protein-1c, adipocyte protein-2, glucose transporter-4 as well as key lipogenic enzymes fatty acid synthase and acetyl-CoA carboxylase, while ERK1/2 activation significantly reversed CUR-reduced lipid accumulation by increasing PPAR-γ. Furthermore, compared with differentiation induced media treated cells, higher dose of CUR (30 μM) significantly decreased the expression of AKT and B-cell lymphoma-2 (BCL-2), while increased the expression of BCL-2-associated X (BAX) and the BAX/BCL-2 expression ratio, suggesting triggered apoptosis by inactivating AKT and increasing BAX/BCL-2 ratio and Caspase-3 expression. Moreover, AKT activation significantly rescued CUR inhibiting lipid accumulation via repressing apoptosis. Conclusion: These results demonstrate that CUR is capable of suppressing differentiation by inhibiting ERK1/2-PPAR-γ signaling pathway and triggering apoptosis via decreasing AKT and subsequently increasing BAX/BCL-2 ratio and Caspase-3, suggesting that CUR provides an important method for the reduction of porcine body fat, as well as the prevention and treatment of human obesity.

키워드

과제정보

We thank Prof. Min Du of Washington State University for critical reading of the manuscript.

참고문헌

  1. Weihrauch-Bluher S, Schwarz P, Klusmann JH. Childhood obesity: increased risk for cardiometabolic disease and cancer in adulthood. Metabolism 2019;92:147-52. https://doi.org/10.1016/j.metabol.2018.12.001
  2. Kopelman PG. Obesity as a medical problem. Nature 2000;404:635-43. https://doi.org/10.1038/35007508
  3. Warnke I, Goralczyk R, Fuhrer E, Schwager J. Dietary constituents reduce lipid accumulation in murine C3H10 T1/2 adipocytes: a novel fluorescent method to quantify fat droplets. Nutr Metab (Lond) 2011;8:30. https://doi.org/10.1186/1743-7075-8-30
  4. Kim HJ, You MK, Lee YH, Kim HJ, Adhikari D, Kim HA. Red pepper seed water extract inhibits preadipocyte differentiation and induces mature adipocyte apoptosis in 3T3-L1 cells. Nutr Res Pract 2018;12:494-502. https://doi.org/10.4162/nrp.2018.12.6.494
  5. Padwal RS, Sharma AM. Prevention of cardiovascular disease: obesity, diabetes and the metabolic syndrome. Can J Cardiol 2010;26 Suppl C(Suppl C):18C-20C. https://doi.org/10.1016/s0828-282x(10)71077-1
  6. Zhang J, Cui YQ, Li X, et al. 5F peptide promotes endothelial differentiation of bone marrow stem cells through activation of ERK1/2 signaling. Eur J Pharmacol 2020;876:173051. https://doi.org/10.1016/j.ejphar.2020.173051
  7. Sun Y, Liu WZ, Liu T, Feng X, Yang N, Zhou HF. Signaling pathway of MAPK/ERK in cell proliferation, differentiation, migration, senescence and apoptosis. J Recept Signal Transduct 2015;35:600-4. https://doi.org/10.3109/10799893.2015.1030412
  8. Ma BC, Xu XY, He S, et al. STC2 modulates ERK1/2 signaling to suppress adipogenic differentiation of human bone marrow mesenchymal stem cells. Biochem Biophys Res Commun 2020;524:163-8. https://doi.org/10.1016/j.bbrc.2020.01.060
  9. Ejaz A, Wu DY, Kwan P, Meydani M. Curcumin inhibits adipogenesis in 3T3-L1 adipocytes and angiogenesis and obesity in C57/BL mice. J Nutr 2009;139:919-25. https://doi.org/10.3945/jn.108.100966
  10. Zhang Y, Huang C. Targeting adipocyte apoptosis: a novel strategy for obesity therapy. Biochem Biophys Res Commun 2012;417:1-4. https://doi.org/10.1016/j.bbrc.2011.11.158
  11. Jin YY, Wang J, Zhang M, et al. Role of bta-miR-204 in the regulation of adipocyte proliferation, differentiation, and apoptosis. J Cell Physiol 2019;234:11037-46. https://doi.org/10.1002/jcp.27928
  12. Srivastava G, Apovian CM. Current pharmacotherapy for obesity. Nat Rev Endocrinol 2018;14:12-24. https://doi.org/10.1038/nrendo.2017.122
  13. Park J, Kim HL, Jung Y, Ahn KS, Kwak HJ, Um JY. Bitter orange (Citrus aurantium Linne) improves obesity by regulating adipogenesis and thermogenesis through AMPK activation. Nutrients 2019;11:1988. https://doi.org/10.3390/nu11091988
  14. Weisberg SP, Leibel R, Tortoriello DV. Dietary curcumin significantly improves obesity-associated inflammation and diabetes in mouse models of diabesity. Endocrinology 2008; 149:3549-58. https://doi.org/10.1210/en.2008-0262
  15. Pan SF, Yang XJ, Jia YM, Li RS, Zhao RQ. Microvesicle-shuttled miR-130b reduces fat deposition in recipient primary cultured porcine adipocytes by inhibiting PPAR-g expression. J Cell Physiol 2014;229:631-9. https://doi.org/10.1002/jcp.24486
  16. Pan SF, Cui YX, Fu ZL, Zhang L, Xing H. MicroRNA-128 is involved in dexamethasone-induced lipid accumulation via repressing SIRT1 expression in cultured pig preadipocytes. J Steroid Biochem Mol Biol 2019;186:185-95. https://doi.org/10.1016/j.jsbmb.2018.10.013
  17. Khan SS, Tarrant M, Kos K, Daly M, Gimbuta C, Farrow CV. Making connections: social identification with new treatment groups for lifestyle management of severe obesity. Clin Psychol Psychother 2020;27:686-96. https://doi.org/10.1002/cpp.2454
  18. Bendixen E, Danielsen M, Hollung K, Gianazza E, Miller I. Farm animal proteomics--a review. J Proteomics 2011;74:282-93. https://doi.org/10.1016/j.jprot.2010.11.005
  19. Dodson MV, Allen RE, Du M, et al. INVITED REVIEW: Evolution of meat animal growth research during the past 50 years: Adipose and muscle stem cells. J Anim Sci 2015;93:457-81. https://doi.org/10.2527/jas.2014-8221
  20. Al Hasan M, Roy P, Dolan S, Martin PE, Patterson S, Bartholomew C. Adhesion G-protein coupled receptor 56 is required for 3T3-L1 adipogenesis. J Cell Physiol 2020;235:1601-14. https://doi.org/10.1002/jcp.29079
  21. Lee MS, Kim Y. Chrysanthemum morifolium flower extract inhibits adipogenesis of 3T3-L1 cells via AMPK/SIRT1 pathway activation. Nutrients 2020;12:2726. https://doi.org/10.3390/nu12092726
  22. Yu HS, Kim WJ, Bae WY, Lee NK, Paik HD. Inula britannica Inhibits Adipogenesis of 3T3-L1 preadipocytes via modulation of mitotic clonal expansion involving ERK 1/2 and Akt signaling pathways. Nutrients 2020;12:3037. https://doi.org/10.3390/nu12103037
  23. Sakuma S, Sumida M, Endoh Y, et al. Curcumin inhibits adipogenesis induced by benzyl butyl phthalate in 3T3-L1 cells. Toxicol Appl Pharmacol 2017;329:158-64. https://doi.org/10.1016/j.taap.2017.05.036
  24. Wang T, Yan RQ, Xu XY, et al. Curcumin represses adipogenic differentiation of human bone marrow mesenchymal stem cells via inhibiting kruppel-like factor 15 expression. Acta Histochem 2019;121:253-9. https://doi.org/10.1016/j.acthis.2018.12.007
  25. Wu LY, Chen CW, Chen LK, Chou HY, Chang CL, Juan CC. Curcumin attenuates adipogenesis by inducing preadipocyte apoptosis and inhibiting adipocyte differentiation. Nutrients 2019;11:2307. https://doi.org/10.3390/nu11102307
  26. Rosen ED, Spiegelman BM. Molecular regulation of adipogenesis. Annu Rev Cell Dev Biol 2000;16:145-71. https://doi.org/10.1146/annurev.cellbio.16.1.145
  27. Wu WJ, Yin YJ, Xu K, Peng YJ, Zhang J. Knockdown of LGALS12 inhibits porcine adipocyte adipogenesis via PKAErk1/2 signaling pathway. Acta Biochim Biophys Sin (Shanghai) 2018;50:960-7. https://doi.org/10.1093/abbs/gmy099
  28. Ferguson BS, Nam H, Morrison RF. Curcumin inhibits 3T3-L1 preadipocyte proliferation by mechanisms involving posttranscriptional p27 regulation. Biochem Biophys Rep 2016;5:16-21. https://doi.org/10.1016/j.bbrep.2015.11.014
  29. Ali AT, Hochfeld WE, Myburgh R, Pepper MS. Adipocyte and adipogenesis. Eur J Cell Biol 2013;92:229-36. https://doi.org/10.1016/j.ejcb.2013.06.001
  30. Farmer SR. Transcriptional control of adipocyte formation. Cell Metab 2006;4:263-73. https://doi.org/10.1016/j.cmet.2006.07.001
  31. Gross DN, Farmer SR, Pilch PF. Glut4 storage vesicles without Glut4: transcriptional regulation of insulin-dependent vesicular traffic. Mol Cell Biol 2004;24:7151-62. https://doi.org/10.1128/MCB.24.16.7151-7162.2004
  32. Hudak CS, Sul HS. Pref-1, a gatekeeper of adipogenesis. Front Endocrinol (Lausanne) 2013;4:79. https://doi.org/10.3389/fendo.2013.00079
  33. Xu B, Ju Y, Song G. Role of p38, ERK1/2, focal adhesion kinase, RhoA/ROCK and cytoskeleton in the adipogenesis of human mesenchymal stem cells. J Biosci Bioeng 2014;117:624-31. https://doi.org/10.1016/j.jbiosc.2013.10.018
  34. Kim J, Han DC, Kim JM, et al. PPAR gamma partial agonist, KR-62776, inhibits adipocyte differentiation via activation of ERK. Cell Mol Life Sci 2009;66:1766-81. https://doi.org/10.1007/s00018-009-9169-4
  35. Kwak DH, Lee JH, Kim T, et al. Aristolochia manshuriensis Kom inhibits adipocyte differentiation by regulation of ERK1/2 and Akt pathway. PLoS One 2012;7:e49530. https://doi.org/10.1371/journal.pone.0049530
  36. Prins JB, O'Rahilly S. Regulation of adipose cell number in man. Clin Sci (Lond) 1997;92:3-11. https://doi.org/10.1042/cs0920003
  37. Nagel SA, Keuper M, Zagotta I, et al. Up-regulation of Bcl-2 during adipogenesis mediates apoptosis resistance in human adipocytes. Mol Cell Endocrinol 2014;382:368-76. https://doi.org/10.1016/j.mce.2013.10.024
  38. Green DR, Reed JC. Mitochondria and apoptosis. Science 1998;281:1309-12. https://doi.org/10.1126/science.281.5381.1309
  39. Morikawa K, Nonaka M, Mochizuki H, Handa K, Hanada H, Hirota K. Naringenin and hesperetin induce growth arrest, apoptosis, and cytoplasmic fat deposit in human preadipocytes. J Agric Food Chem 2008;56:11030-7. https://doi.org/10.1021/jf801965n
  40. Reusch JE, Klemm DJ. Inhibition of cAMP-response elementbinding protein activity decreases protein kinase B/Akt expression in 3T3-L1 adipocytes and induces apoptosis. J Biol Chem 2002;277:1426-32. https://doi.org/10.1074/jbc.M107923200
  41. Xiao YY, Yuan TC, Yao WQ, Liao K. 3T3-L1 adipocyte apoptosis induced by thiazolidinediones is peroxisome proliferatoractivated receptor-gamma-dependent and mediated by the caspase-3-dependent apoptotic pathway. FEBS J 2010;277:687-96. https://doi.org/10.1111/j.1742-4658.2009.07514.x