Curcumin represses lipid accumulation through inhibiting ERK1/2-PPAR-γ signaling pathway and triggering apoptosis in porcine subcutaneous preadipocytes
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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 (BCL2), 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.
Abstract
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 (BCL2), 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.
참고문헌 (41)
- Weihrauch-Blüher S, "Childhood obesity : increased risk for cardiometabolic disease and cancer in adulthood", Metabolism, 92, pp.147~152 (2019)[DOI]
- Kopelman PG, "Obesity as a medical problem", Nature, 404, pp.635~643 (2000)[DOI]
- Warnke I, "Dietary constituents reduce lipid accumulation in murine C3H10 T1", 2adipocytes: a novel fluorescent method to quantify fat droplets, 2011, pp.8, 30~
- 김화진, "Red pepper seed water extract inhibits preadipocyte differentiation and induces mature adipocyte apoptosis in 3T3-L1 cells", Nutrition Research and Practice, 12(6), pp.494~502 (2018), 한국영양학회
- Padwal RS, "Prevention of cardiovascular disease : obesity, diabetes and the metabolic syndrome", Can J Cardiol, 26 Suppl C(Suppl C), pp.18C~20C (2010)[DOI]
- Zhang J, "5F peptide promotes endothelial differentiation of bone marrow stem cells through activation of ERK1", 2 signaling, 2020, pp.876, 173051~
- Sun Y, "Signaling pathway of MAPK", ERK in cell proliferation, differentiation, migration, senescence and apoptosis, 2015, pp.35, 600~604
- Ma BC, "STC2 modulates ERK1", 2 signaling to suppress adipogenic differentiation of human bone marrow mesenchymal stem cells, 2020, pp.524, 163~168
- Ejaz A, "Curcumin inhibits adipogenesis in 3T3-L1 adipocytes and angiogenesis and obesity in C57", BL mice, 2009, pp.139, 919~925
- Zhang Y, "Targeting adipocyte apoptosis : a novel strategy for obesity therapy", Biochem Biophys Res Commun, 417, pp.1~4 (2012)[DOI]
- Jin YY, "Role of bta-miR-204 in the regulation of adipocyte proliferation, differentiation, and apoptosis", J Cell Physiol, 234, pp.11037~11046 (2019)[DOI]
- Srivastava G, "Current pharmacotherapy for obesity", Nat Rev Endocrinol, 14, pp.12~24 (2018)[DOI]
- Park J, "Bitter orange (Citrus aurantium Linné) improves obesity by regulating adipogenesis and thermogenesis through AMPK activation", Nutrients, 11, pp.1988~ (2019)[DOI]
- Weisberg SP, "Dietary curcumin significantly improves obesity-associated inflammation and diabetes in mouse models of diabesity", Endocrinology, 149, pp.3549~3558 (2008)[DOI]
- Pan SF, "Microvesicle-shuttled miR-130b reduces fat deposition in recipient primary cultured porcine adipocytes by inhibiting PPAR-g expression", J Cell Physiol, 229, pp.631~639 (2014)[DOI]
- Pan SF, "MicroRNA-128 is involved in dexamethasone-induced lipid accumulation via repressing SIRT1 expression in cultured pig preadipocytes", J Steroid Biochem Mol Biol, 186, pp.185~195 (2019)[DOI]
- Khan SS, "Making connections : social identification with new treatment groups for lifestyle management of severe obesity", Clin Psychol Psychother, 27, pp.686~696 (2020)[DOI]
- Bendixen E, "Farm animal proteomics--a review", J Proteomics, 74, pp.282~293 (2011)[DOI]
- Dodson MV, "INVITED REVIEW : Evolution of meat animal growth research during the past 50 years : Adipose and muscle stem cells", J Anim Sci, 93, pp.457~481 (2015)[DOI]
- Al Hasan M, "Adhesion G-protein coupled receptor 56 is required for 3T3-L1 adipogenesis", J Cell Physiol, 235, pp.1601~1614 (2020)[DOI]
- Lee MS, "Chrysanthemum morifolium flower extract inhibits adipogenesis of 3T3-L1 cells via AMPK", SIRT1 pathway activation, 2020, pp.12, 2726~
- Yu HS, "Inula britannica Inhibits Adipogenesis of 3T3-L1 preadipocytes via modulation of mitotic clonal expansion involving ERK 1", 2 and Akt signaling pathways, 2020, pp.12, 3037~
- Sakuma S, "Curcumin inhibits adipogenesis induced by benzyl butyl phthalate in 3T3-L1cells", Toxicol Appl Pharmacol, 329, pp.158~164 (2017)[DOI]
- Wang T, "Curcumin represses adipogenic differentiation of human bone marrow mesenchymal stem cells via inhibiting kruppel-like factor 15 expression", Acta Histochem, 121, pp.253~259 (2019)[DOI]
- Wu LY, "Curcumin attenuates adipogenesis by inducing preadipocyte apoptosis and inhibiting adipocyte differentiation", Nutrients, 11, pp.2307~ (2019)[DOI]
- Rosen ED, "Molecular regulation of adipogenesis", Annu Rev Cell Dev Biol, 16, pp.145~171 (2000)[DOI]
- Wu WJ, "Knockdown of LGALS12 inhibits porcine adipocyte adipogenesis via PKAErk1", 2 signaling pathway, 2018, pp.50, 960~967
- Ferguson BS, "Curcumin inhibits 3T3-L1preadipocyte proliferation by mechanisms involving posttranscriptional p27 regulation", Biochem Biophys Rep, 5, pp.16~21 (2016)[DOI]
- Ali AT, "Adipocyte and adipogenesis", Eur J Cell Biol, 92, pp.229~236 (2013)[DOI]
- Farmer SR, "Transcriptional control of adipocyte formation", Cell Metab, 4, pp.263~273 (2006)[DOI]
- Gross DN, "Glut4 storage vesicles without Glut4 : transcriptional regulation of insulin-dependent vesicular traffic", Mol Cell Biol, 24, pp.7151~7162 (2004)[DOI]
- Hudak CS, "Pref-1, a gatekeeper of adipogenesis", Front Endocrinol(Lausanne), 4, pp.79~ (2013)[DOI]
- Xu B, "Role of p38, ERK1", 2, focal adhesion kinase, RhoA, J Biosci Bioeng, pp.2014, 117[DOI]
- Kim J, "PPAR gamma partial agonist, KR-62776, inhibits adipocyte differentiation via activation of ERK", Cell Mol Life Sci, 66, pp.1766~1781 (2009)[DOI]
- Kwak DH, "Aristolochia manshuriensis Kom inhibits adipocyte differentiation by regulation of ERK1", 2and Akt pathway, 2012, pp.7, e49530~
- Prins JB, "Regulation of adipose cell number in man", Clin Sci(Lond), 92, pp.3~11 (1997)[DOI]
- Nagel SA, "Up-regulation of Bcl-2during adipogenesis mediates apoptosis resistance in human adipocytes", Mol Cell Endocrinol, 382, pp.368~376 (2014)[DOI]
- Green DR, "Mitochondria and apoptosis", Science, 281, pp.1309~1312 (1998)[DOI]
- Morikawa K, "Naringenin and hesperetin induce growth arrest, apoptosis, and cytoplasmic fat deposit in human preadipocytes", J Agric Food Chem, 56, pp.11030~11037 (2008)[DOI]
- Reusch JE, "Inhibition of cAMP-response elementbinding protein activity decreases protein kinase B", Akt expression in 3T3-L1 adipocytes and induces apoptosis, 2002, pp.277, 1426~1432
- Xiao YY, "3T3-L1 adipocyte apoptosis induced by thiazolidinediones is peroxisome proliferatoractivated receptor-gamma-dependent and mediated by the caspase-3-dependent apoptotic pathway", FEBS J, 277, pp.687~696 (2010)[DOI]