BibTex RIS Kaynak Göster

Effect of capsaicin on transcription factors in 3T3-L1 cell line

Yıl 2015, Cilt: 20 Sayı: 1, 34 - 45, 02.06.2015

Öz

Capsaicin is a spicy ingredient of Capsicum annuum and a lipophilic, crystalline, odorless and colorless alkaloid. Although the effect of capsaicin on adipocyte differentiation is well-known, the role of capsaicin on transcription factors while adipocyte differentiation is not clear. The aim of this study is thus to identify and characterize the transcription factors in the process of adipocyte differentiation after the capsaicin treatment. In this study, concentration of 0, 50, 100, 150, 200 and 250 µM capsaicin were treated to 3T3-L1 pre-adipocytes in cell culture. MTT cell cytotoxicity, cell viability with trypan blue staining, Lactate Dehydrogenase (LDH) enzyme assay, triglyceride content assay, Glycerol-3-Phosphate Dehydrogenase (GPDH) activity, Oil Red O staining and mRNA levels of transcription factors (PPARγ, C/EBPα and SREBP-l) were invastigated in capsaicin induced 3T3-L1 preadipocyte cell line. Capsaicin treatment decreased cell population growth of 3T3-L1 preadipocytes, assessed with trypan blue staining, MTT test and rising of LDH release proportion. Capsaicin inhibited GPDH activity and intracellular triglyceride content in 3T3-L1 adipocytes in all treated .groups in a dose-dependent manner. Oil Red O staining indicated that capsaicin inhibited adipocyte differentiation in 3T3-L1 adipocytes in all treatment groups. In this study, it was revealed that exposing 3T3-L1 preadipocytes and differentiating postconfluent preadipocytes to different doses of capsaicin decreased PPARγ, C/EBPα and SREBP-1 mRNA levels as compared with their controls without treatment in dose dependent manner. Although, reduction of PPARγ mRNA level was statistical significant, this decrease was not significant in C/EBPα and SREBP-1 mRNA levels. This study demonstrated that capsaicin treatment inhibited the adipogenesis through the down-regulation of transcription factors, especially PPARγ. Alternative mechanisms may involve cell cycle arrest and the induction of apoptosis. Since capsaicin is the main component found in hot pepper, the consumption of hot pepper may contribute to the maintenance of body weight and prevent the development of obesity.

 

Keywords: Capsaicin, 3T3-L1, adipocyte, transcription factors, PPARγ, C/EBPα, SREBP-l

Kaynakça

  • Duncan RE, Ahmadian M, Jaworski K, Sarkadi-Nagy E, Sul HS. Regulation of lipolysis in adipocytes. Annu Rev Nutr 2007; 27: 79-101.
  • MacDougald OA1, Lane MD. Adipocyte differentiation. When precursors are also regulators. Curr Biol 1995; 5: 618-621.
  • Novakofski J. Adipogenesis: usefulness of in vitro and in vivo experimental models. J Anim Sci 2004; 82: 905-915.
  • Schoonjans K, Staels B, Auwerx J. The peroxisome proliferator activated receptors (PPARS) and their effects on lipid metabolism and adipocyte differentiation. Biochim Biophys Acta 1996; 1302: 93-109.
  • Shimano H, Yahagi N, Amemiya-Kudo M, et al. Sterol regulatory element-binding protein-1 as a key transcription factor for nutritional induction of lipogenic enzyme genes. J Biol Chem 1999; 274: 35832-35839.
  • Escher P, Wahli W. Peroxisome proliferator-activated receptors: insight into multiple cellular functions. Mutat Res 2000; 448: 121-138.
  • Feige JN1, Gelman L, Michalik L, Desvergne B, Wahli W. From molecular action to physiological outputs: peroxisome proliferator-activated receptors are nuclear receptors at the crossroads of key cellular functions. Prog Lipid Res 2006; 45: 120-159.
  • Tontonoz P, Hu E, Spiegelman BM. Regulation of adipocyte gene expression and differentiation by peroxisome proliferator activated receptor gamma. Curr Opin Genet Dev 1995; 5: 571-576.
  • Cao Z, Umek RM, McKnight SL. Regulated expression of three C/EBP isoforms during adipose conversion of 3T3-L1 cells. Genes Dev 1991; 5: 1538- 1552.
  • Tang QQ, Otto TC, Lane MD. CCAAT/enhancer- binding protein beta is required for mitotic clonal expansion during adipogenesis. Proc Natl Acad Sci USA 2003; 100: 850-855.
  • Zhang JW, Klemm DJ, Vinson C, Lane MD. Role of CREB in transcriptional regulation of CCAAT/enhancer-binding protein beta gene during adipogenesis. J Biol Chem 2004; 279: 4471-4478.
  • Bertile F, Raclot T. mRNA levels of SREBP-1c do not coincide with the changes in adipose lipogenic gene expression. Biochem Biophys Res Commun 2004; 325: 827-834.
  • Matsuda M, Korn BS, Hammer RE, et al. SREBP cleavage-activating protein (SCAP) is required for increased lipid synthesis in liver induced by cholesterol deprivation and insulin elevation. Genes Dev 2001; 15: 1206-1216.
  • Sekiya M, Yahagi N, Matsuzaka T, et al. SREBP-1- independent regulation of lipogenic gene expression in adipocytes. J Lipid Res 2007; 48: 1581-1591.
  • Bernlohr DA, Bolanowski MA, Kelly TJ Jr, Lane MD. Evidence for an increase in transcription of specific mRNAs during differentiation of 3T3-L1 preadipocytes. J Biol Chem 1985; 260: 5563-5567.
  • Student AK, Hsu RY, Lane MD. Induction of fatty acid synthetase synthesis in differentiating 3T3-L1 preadipocytes. J Biol Chem 1980; 255: 4745-4750.
  • Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983; 65: 55-63.
  • Sanford WC. A new method for dispersing strongly adhesive cells in tissue culture. In Vitro 1974; 10: 281-283.
  • Satory DL, Smith SB. Conjugated linoleic acid inhibits proliferation but stimulates lipid filling of murine 3T3-L1 preadipocytes. J Nutr 1999; 129: 92- 97.
  • Ramírez-Zacarías JL, Castro-Muñozledo F, Kuri- Harcuch W. Quantitation of adipose conversion and triglycerides by staining intracytoplasmic lipids with Oil red O. Histochemistry 1992; 97: 493-497.
  • Wise LS, Green H. Participation of one isozyme of cytosolic glycerophosphate dehydrogenase in the adipose conversion of 3T3 cells. J Biol Chem 1979; 254: 273-275.
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72: 248-254.
  • Wang J. DNA/RNA Isolation and Quantitation. In: Hu P, Hegde M, Lennon PA. (eds). Modern Clinical Molecular Techniques (1st ed.). Springer-Verlag, New York Heidelberg Dordrecht London, 2012, pp 11-22
  • He Q, Huang C, Zhao L, et al. α-Naphthoflavone inhibits 3T3-L1 pre-adipocytes differentiation via modulating p38MAPK signaling. Int J Clin Exp Pathol 2013; 6: 168-178.
  • Tirard J, Gout J, Lefrançois-Martinez AM, et al. A novel inhibitory protein in adipose tissue, the aldo- keto reductase AKR1B7: its role in adipogenesis. Endocrinology 2007; 148: 1996-2005.
  • Wang YW, Jones PJ. Conjugated linoleic acid and obesity control: efficacy and mechanisms. Int J Obes Relat Metab Disord 2004; 28: 941-955.
  • Moro CO, Basile G. Obesity and medicinal plants. Fitoterapia 2000; 71: 73-82.
  • Heber D. Herbal preparations for obesity: are they useful? Prim Care 2003; 30: 441-463.
  • Baboota RK, Singh DP, Sarma SM, et al. Capsaicin induces "brite" phenotype in differentiating 3T3-L1 preadipocytes. PLoS One 2014; 9: 103093.
  • Feng Z, Hai-ning Y, Xiao-man C, et al. Effect of yellow capsicum extract on proliferation and differentiation of 3T3-L1 preadipocytes. Nutrition 2014; 30: 319-325.
  • Lee MS, Kim CT, Kim IH, Kim Y. Effects of capsaicin on lipid catabolism in 3T3-L1 adipocytes. Phytother Res 2011; 25: 935-939.
  • Han J, Isoda H, Maekawa T. Analysis of the mechanism of the tight-junctional permeability increase by capsaicin treatment on the intestinal Caco- 2 cells. Cytotechnology 2002; 40: 93-98.
  • Ahn IS, Do MS, Kim SO, et al. Antiobesity effect of Kochujang (Korean fermented red pepper paste) extract in 3T3-L1 adipocytes. J Med Food 2006; 9: 15-21.
Yıl 2015, Cilt: 20 Sayı: 1, 34 - 45, 02.06.2015

Öz

Kaynakça

  • Duncan RE, Ahmadian M, Jaworski K, Sarkadi-Nagy E, Sul HS. Regulation of lipolysis in adipocytes. Annu Rev Nutr 2007; 27: 79-101.
  • MacDougald OA1, Lane MD. Adipocyte differentiation. When precursors are also regulators. Curr Biol 1995; 5: 618-621.
  • Novakofski J. Adipogenesis: usefulness of in vitro and in vivo experimental models. J Anim Sci 2004; 82: 905-915.
  • Schoonjans K, Staels B, Auwerx J. The peroxisome proliferator activated receptors (PPARS) and their effects on lipid metabolism and adipocyte differentiation. Biochim Biophys Acta 1996; 1302: 93-109.
  • Shimano H, Yahagi N, Amemiya-Kudo M, et al. Sterol regulatory element-binding protein-1 as a key transcription factor for nutritional induction of lipogenic enzyme genes. J Biol Chem 1999; 274: 35832-35839.
  • Escher P, Wahli W. Peroxisome proliferator-activated receptors: insight into multiple cellular functions. Mutat Res 2000; 448: 121-138.
  • Feige JN1, Gelman L, Michalik L, Desvergne B, Wahli W. From molecular action to physiological outputs: peroxisome proliferator-activated receptors are nuclear receptors at the crossroads of key cellular functions. Prog Lipid Res 2006; 45: 120-159.
  • Tontonoz P, Hu E, Spiegelman BM. Regulation of adipocyte gene expression and differentiation by peroxisome proliferator activated receptor gamma. Curr Opin Genet Dev 1995; 5: 571-576.
  • Cao Z, Umek RM, McKnight SL. Regulated expression of three C/EBP isoforms during adipose conversion of 3T3-L1 cells. Genes Dev 1991; 5: 1538- 1552.
  • Tang QQ, Otto TC, Lane MD. CCAAT/enhancer- binding protein beta is required for mitotic clonal expansion during adipogenesis. Proc Natl Acad Sci USA 2003; 100: 850-855.
  • Zhang JW, Klemm DJ, Vinson C, Lane MD. Role of CREB in transcriptional regulation of CCAAT/enhancer-binding protein beta gene during adipogenesis. J Biol Chem 2004; 279: 4471-4478.
  • Bertile F, Raclot T. mRNA levels of SREBP-1c do not coincide with the changes in adipose lipogenic gene expression. Biochem Biophys Res Commun 2004; 325: 827-834.
  • Matsuda M, Korn BS, Hammer RE, et al. SREBP cleavage-activating protein (SCAP) is required for increased lipid synthesis in liver induced by cholesterol deprivation and insulin elevation. Genes Dev 2001; 15: 1206-1216.
  • Sekiya M, Yahagi N, Matsuzaka T, et al. SREBP-1- independent regulation of lipogenic gene expression in adipocytes. J Lipid Res 2007; 48: 1581-1591.
  • Bernlohr DA, Bolanowski MA, Kelly TJ Jr, Lane MD. Evidence for an increase in transcription of specific mRNAs during differentiation of 3T3-L1 preadipocytes. J Biol Chem 1985; 260: 5563-5567.
  • Student AK, Hsu RY, Lane MD. Induction of fatty acid synthetase synthesis in differentiating 3T3-L1 preadipocytes. J Biol Chem 1980; 255: 4745-4750.
  • Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983; 65: 55-63.
  • Sanford WC. A new method for dispersing strongly adhesive cells in tissue culture. In Vitro 1974; 10: 281-283.
  • Satory DL, Smith SB. Conjugated linoleic acid inhibits proliferation but stimulates lipid filling of murine 3T3-L1 preadipocytes. J Nutr 1999; 129: 92- 97.
  • Ramírez-Zacarías JL, Castro-Muñozledo F, Kuri- Harcuch W. Quantitation of adipose conversion and triglycerides by staining intracytoplasmic lipids with Oil red O. Histochemistry 1992; 97: 493-497.
  • Wise LS, Green H. Participation of one isozyme of cytosolic glycerophosphate dehydrogenase in the adipose conversion of 3T3 cells. J Biol Chem 1979; 254: 273-275.
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72: 248-254.
  • Wang J. DNA/RNA Isolation and Quantitation. In: Hu P, Hegde M, Lennon PA. (eds). Modern Clinical Molecular Techniques (1st ed.). Springer-Verlag, New York Heidelberg Dordrecht London, 2012, pp 11-22
  • He Q, Huang C, Zhao L, et al. α-Naphthoflavone inhibits 3T3-L1 pre-adipocytes differentiation via modulating p38MAPK signaling. Int J Clin Exp Pathol 2013; 6: 168-178.
  • Tirard J, Gout J, Lefrançois-Martinez AM, et al. A novel inhibitory protein in adipose tissue, the aldo- keto reductase AKR1B7: its role in adipogenesis. Endocrinology 2007; 148: 1996-2005.
  • Wang YW, Jones PJ. Conjugated linoleic acid and obesity control: efficacy and mechanisms. Int J Obes Relat Metab Disord 2004; 28: 941-955.
  • Moro CO, Basile G. Obesity and medicinal plants. Fitoterapia 2000; 71: 73-82.
  • Heber D. Herbal preparations for obesity: are they useful? Prim Care 2003; 30: 441-463.
  • Baboota RK, Singh DP, Sarma SM, et al. Capsaicin induces "brite" phenotype in differentiating 3T3-L1 preadipocytes. PLoS One 2014; 9: 103093.
  • Feng Z, Hai-ning Y, Xiao-man C, et al. Effect of yellow capsicum extract on proliferation and differentiation of 3T3-L1 preadipocytes. Nutrition 2014; 30: 319-325.
  • Lee MS, Kim CT, Kim IH, Kim Y. Effects of capsaicin on lipid catabolism in 3T3-L1 adipocytes. Phytother Res 2011; 25: 935-939.
  • Han J, Isoda H, Maekawa T. Analysis of the mechanism of the tight-junctional permeability increase by capsaicin treatment on the intestinal Caco- 2 cells. Cytotechnology 2002; 40: 93-98.
  • Ahn IS, Do MS, Kim SO, et al. Antiobesity effect of Kochujang (Korean fermented red pepper paste) extract in 3T3-L1 adipocytes. J Med Food 2006; 9: 15-21.
Toplam 33 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Bölüm Articles
Yazarlar

Mehmet Berköz

Metin Yildirim Bu kişi benim

Gulhan Arvas Bu kişi benim

Omer Turkmen Bu kişi benim

Oruc Allahyerdiyev Bu kişi benim

Yayımlanma Tarihi 2 Haziran 2015
Yayımlandığı Sayı Yıl 2015 Cilt: 20 Sayı: 1

Kaynak Göster

APA Berköz, M., Yildirim, M., Arvas, G., Turkmen, O., vd. (2015). Effect of capsaicin on transcription factors in 3T3-L1 cell line. EASTERN JOURNAL OF MEDICINE, 20(1), 34-45.
AMA Berköz M, Yildirim M, Arvas G, Turkmen O, Allahyerdiyev O. Effect of capsaicin on transcription factors in 3T3-L1 cell line. EASTERN JOURNAL OF MEDICINE. Haziran 2015;20(1):34-45.
Chicago Berköz, Mehmet, Metin Yildirim, Gulhan Arvas, Omer Turkmen, ve Oruc Allahyerdiyev. “Effect of Capsaicin on Transcription Factors in 3T3-L1 Cell Line”. EASTERN JOURNAL OF MEDICINE 20, sy. 1 (Haziran 2015): 34-45.
EndNote Berköz M, Yildirim M, Arvas G, Turkmen O, Allahyerdiyev O (01 Haziran 2015) Effect of capsaicin on transcription factors in 3T3-L1 cell line. EASTERN JOURNAL OF MEDICINE 20 1 34–45.
IEEE M. Berköz, M. Yildirim, G. Arvas, O. Turkmen, ve O. Allahyerdiyev, “Effect of capsaicin on transcription factors in 3T3-L1 cell line”, EASTERN JOURNAL OF MEDICINE, c. 20, sy. 1, ss. 34–45, 2015.
ISNAD Berköz, Mehmet vd. “Effect of Capsaicin on Transcription Factors in 3T3-L1 Cell Line”. EASTERN JOURNAL OF MEDICINE 20/1 (Haziran 2015), 34-45.
JAMA Berköz M, Yildirim M, Arvas G, Turkmen O, Allahyerdiyev O. Effect of capsaicin on transcription factors in 3T3-L1 cell line. EASTERN JOURNAL OF MEDICINE. 2015;20:34–45.
MLA Berköz, Mehmet vd. “Effect of Capsaicin on Transcription Factors in 3T3-L1 Cell Line”. EASTERN JOURNAL OF MEDICINE, c. 20, sy. 1, 2015, ss. 34-45.
Vancouver Berköz M, Yildirim M, Arvas G, Turkmen O, Allahyerdiyev O. Effect of capsaicin on transcription factors in 3T3-L1 cell line. EASTERN JOURNAL OF MEDICINE. 2015;20(1):34-45.