EFFECT OF TROGLITAZONE ON MORPHOLOGICAL CHANGES AND PPAR-γ GENE EXPRESSION IN 3T3-L1 ADIPOCYTE CELLS
Article Sidebar
Main Article Content
Abstract
3T3-L1 cells are extensively used as a model to study adipogenesis. However, one major concern is the prolonged period of time it takes the cells to differentiate into adipocytes form. To induce this differentiation, the adipogenic induction media is required. In this study, troglitazone, a hypoglycemic agent was added to adipogenic induction media and observed in order to determine the morphological changes and peroxisome proliferator-activated receptor gamma (PPAR-γ) gene expression in 3T3-L1 differentiation. It is generally known that PPAR-ê© plays an important role as a transcription factor in adipocyte differentiation. Based on Oil Red O Staining, adipogenic induction with or without troglitazone changed the 3T3-L1 pre-adipocytes into mature round fat cells characterized by red droplet lipids. This cell also had a high absorbance level and degree of droplet accumulation of P≤ 0.05 in each group. In addition, cells treated by troglitazone had the highest PPAR-ê© mRNA level (1.9 fold) than those treated by adipogenic induction media without troglitazone or cells un-treated at all.
Article Details
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
Authors who publish with this journal agree to the following terms:
a). Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Attribution-NonCommercial-ShareAlike 4.0 International that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
b). Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
c). Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).
References
Ali AT, Hochfeld WE, Myburgh R, Pepper MS (2013) Adipocyte and adipogenesis. Eur J Cell Biol 92:229-236. doi: 10.1016/j.ejcb.2013.06.001
Alvim RO, Cheuhen MR, Machado SR, Sousa AGP, Santos PCJL (2015) General aspects of muscle glucose uptake. Ann Braz Acad Sci 87:351-368. doi: 10.1590/0001-3765201520140225
Aoyagi R, Tago M, Fujiwara Y, Tamura H (2014) Coffee inhibits adipocyte differentiation via inactivation of PPARγ. Biol Pharm Bull 37:1820-1825. doi: 10.1248/bpb.b14-00378
Camp HS, Li O, Wise SC, Hong YH, Frankowski CL, Shen X, Vanbogelen, Leff T (2000) Differential activation of peroxisome proliferator-activated receptor-γ by troglitazone and rosiglitazone. Diabetes 49:539-547. doi: 10.2337/diabetes.49.4.539
Ciaraldi TP, Kong APS, Chu NV, Kim DD, Baxi S, Loviscach M, Plodkowski R, Reitz R, Coulfield M, Mudaliar S, Henry RR (2002) Regulation of glucose transport and insulin signaling by troglitazone or metformin in adipose tissue of type 2 diabetic subject. Diabetes 51:30-36. doi:10.2337/diabetes.51.1.30
Deng T, Sieglaff DH, Zang A, Lyon CJ, Ayers SD, Cvoro A, Gupte AA, Xia X, Baxter JD, Webb P, Hsueh WA (2011) A peroxisome proliferator-activated receptor gamma (PPARγ)/PPARγ coactivator 1β autoregulatory loop in adipocyte mitochondrial function. J Biol Chem 286:30723-30731. doi: 10.1074/jbc.M111.251926
Ghoniem AA, Acil Y, Wiltfang J, Gierloff M (2015) Improved adipogenic in vitro differentiation: Comparison of different adipogenic cell culture media on human fat and bone stroma cells for fat tissue engineering. Anat Cell Biol 48:85-94. doi: 10.5115/acb.2015.48.2.85
Gregoire FM, Smas CM, Sul HS (1998) Understanding adipocyte differentiation. Physiol Rev 78:783-809. doi: 10.1152/physrev.1998.78.3.783
Hamm JK, Park BH, Farmer SR (2001) A role for C/EBPβ in regulating peroxisome proliferator-activated receptor γ activity during adipogenesis in 3T3-L1 preadipocytes. J Biol Chem 276:18464-18471. doi: 10.1074/jbc.M100797200
Kang JW, Nam D, Kim KH, Huh JE, Lee JD (2013) Effect of gambisan on the Inhibition of adipogenesis in 3T3-L1 adipocytes. Evidence-Based Complementary Altern Med 2013:1-11. doi: 10.1155/2013/789067
Kawahara A, Haraguchi N, Tsuchiya H, Ikeda Y, Hama S, Kogure K (2013) Peroxisome proliferator-activated receptor γ (PPARγ)-independent specific cytotoxicity against Immature adipocytes induced by PPARγ antagonist T0070907. Biol Pharm Bull 36:1428-1434. doi: 10.1248/bpb.b13-00024
Lasar D, Rosenwald M, Kiehlmann E, Balaz M, Tall B, Opitz L, Lidell ME, Zamboni N, Krznar P, Sun W, Varga L, Stefanicka P, Ukropec J, Nuutila P, Virtanen K, Amri EZ, Enerback S, Wahli W, Wolfrum (2018) Peroxisome proliferator activated receptor gamma controls mature brown adipocyte inducibility through glycerol kinase. Cell Rep 22:760-773. doi: 10.1016/j.celrep.2017.12.067
Lee JE, Ge K (2014) Transcriptional and epigenetic regulation of PPARγ expression during adipogenesis. Cell Biosci 4:1-11. doi: 10.1186/2045-3701-4-29
Martinez L, Berenguer M, Bruce MC, Le Marchand-Brustel Y, Govers R (2010) Rosiglitazone increases cell surface GLUT4 levels in 3T3-L1 adipocytes through an enhancement of endosomal recycling. Biochem Pharmacol 79:1300–1309. doi: 10.1016/j.bcp.2009.12.013
Mehra A, MacDonald I, Pillay TS (2007) Variability in 3T3-L1 adipocyte differentiation depending on cell culture dish. Anal Biochem 362:281-283. doi: 10.1016/j.ab.2006.12.016
Min B, Lee H, Song JH, Han MJ, Chung J (2014) Arctiin inhibits adipogenesis in 3T3-L1 cells and decreases adiposity and body weight in mice fed a high-fat diet. Nutr Res Pract 8:655-661. doi: 10.4162/nrp.2014.8.6.655
Morrison S, McGee SL (2015) 3T3-L1 adipocytes display phenotypic characteristics of multiple adipocyte lineages. Adipocyte 4:295–302. doi: 10.1080/21623945.2015.1040612
Rieusset J, Andreelli F, Auboeuf D, Roques M, Vallier P, Riou JP, Auwerx J, Laville M, Vidal H (1999) Insulin acutely regulates the expression of the peroxisome proliferator-activated receptor-γ in human adipocytes. Diabetes 48:699-705. doi: 10.2337/diabetes.48.4.699
Ruiz-Ojeda FJ, Ruperez AI, Gomez-Llorente C, Gil A, Aquilera CM (2016) Cell models and their application for studying adipogenic differentiation in relation to obesity: A Review. Int J Mol Sci 17:1-26. doi: 10.3390/ijms17071040
Siersbæk R, Nielsen R, Mandrup S (2010) PPARγ in adipocyte differentiation and metabolism - novel insights from genome-wide studies. FEBS Lett 584:3242-3249. doi: 10.1016/j.febslet.2010.06.010
Taher M, Amiroudine MZAM, Zakaria TMFST, Susanti D, Ichwan SJA, Kaderi MA, Ahmed QU, Zakaria ZA (2015) α-Mangostin improves glucose uptake and inhibits adipocytes differentiation in 3T3-L1 cells via PPAR-γ, GLUT4, and leptin expressions. Evidence-Based Complementary Altern Med 2015:1-9. doi: 10.1155/2015/740238
Vinayagam R, Xu B (2015) Antidiabetic properties of dietary flavonoids: A cellular mechanism review. Nutr Metab 12:60. doi: 10.1186/s12986-015-0057-7
Vishwanath D, Srinivasan H, Patil MS, Seetarama S, Agrawal SK, Dixit MN, Dhar K (2013) Novel method to differentiate 3T3 L1 cells in vitro to produce highly sensitive adipocytes for a GLUT4 mediated glucose uptake using fluorescent glucose analog. J Cell Commun Signal 7:129140. doi: 10.1007/s12079-012-0188-9
Zebisch K, Voigt V, Wabitsch M, Brandch M (2012) Protocol for effective differentiation of 3T3-L1 cells to adipocytes. Anal Biochem 425:88-90. doi: 10.1016/j.ab.2012.03.005
Zhang J, Tang H, Zhang Y, Deng R, Shao L, Liu Y, Li F, Wang X, Zhou L (2014) Identification of suitable reference genes for quantitative RT-PCR during 3T3-L1 adipocyte differentiation 33:1209-1218. doi: 10.3892/ijmm.2014.1695
Zhu HY, Bai WD, Li J, Tao K, Wang HT, Yang XK, Liu JQ, Wang YC, He T, Xie ST, Hu DH (2016) Peroxisome proliferator-activated receptor-γ agonist troglitazone suppresses transforming growth factor-β1 signalling through miR-92b upregulation-inhibited Axl expression in human keloid fibroblasts in vitro. Am J Transl Res 8:3460-3470. PMID: https://www.ncbi.nlm.nih.gov/pubmed/27648136">27648136