EFFECT OF TROGLITAZONE ON MORPHOLOGICAL CHANGES AND PPAR-γ GENE EXPRESSION IN 3T3-L1 ADIPOCYTE CELLS

Asri Sulfianti, Mayriska Triwulansari, . Nuralih, . Churiyah

Abstract


Efek Troglitazone terhadap Perubahan Morfologi dan Ekspresi Gen PPAR- γ di Dalam Sel Adiposa 3T3-L1

 

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-ꝩ 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-ꝩ mRNA level (1.9 fold) than those treated by adipogenic induction media without troglitazone or cells un-treated at all. 

Keywords: 3T3-L1, adipocyte, differentiation, PPAR-ꝩ, troglitazone

 

ABSTRAK

Sel 3T3-L1 adalah jenis sel yang banyak digunakan dalam studi adipogenesis. Namun, salah satu kelemahan sel tersebut adalah lamanya waktu yang dibutuhkan bagi sel pre-adiposa untuk berdiferensiasi menjadi sel adiposa. Selain itu, dibutuhkan pula media induksi khusus untuk mengubah sel menjadi sel adiposa. Pada penelitian ini, kami mengobservasi fungsi troglitazone, sebagai antidiabetes terhadap perubahan morfologi dan ekspresi gen peroxisome proliferator-activated receptor gamma (PPAR-γ). Telah diketahui bahwa PPAR-ꝩ berperan penting sebagai factor transkripsi dalam diferensasi sel adiposa. Berdasarkan pewarnaan ORO, induksi sel pre-adiposa 3T3-L1 dengan media induksi dengan dan tanpa troglitazone merubah sel preadiposa menjadi sel berbentuk bulat yang dikarakterisasi dengan akumulasi droplet lemak. Nilai absorbansi sel adiposa juga menandakan adanya perbedaan yang signifikan antara kelompok sel yang diberi troglitazone dan tidak, dan sel tanpa diberi media induksi. Sementara, pada kelompok sel yang diberi troglitazone memiliki ekspresi mRNA PPAR-ꝩ (1,9 kali) tertinggi jika dibandingkan dengan sel yang diberi media induksi tanpa troglitazone, dan tanpa media induksi sama sekali.

Kata Kunci: 3T3-L1, adiposa, diferensiasi, PPAR-ꝩ, troglitazone


Keywords


3T3-L1; adipocyte; differentiation; PPAR-ꝩ; troglitazone

Full Text:

PDF

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: 27648136




DOI: http://dx.doi.org/10.29122/jbbi.v6i1.3117

Refbacks

  • There are currently no refbacks.


© Copyright: BY-NC-SA