Hostname: page-component-78c5997874-8bhkd Total loading time: 0 Render date: 2024-11-19T08:39:10.498Z Has data issue: false hasContentIssue false

Effect of retinoid X receptor α (RXRα) on apoptosis in primary cultured porcine pre-adipocytes

Published online by Cambridge University Press:  29 January 2010

Lin Ya-Qiu
Affiliation:
Laboratory of Animal Fat Deposition and Muscle Development, Northwest Agriculture and Forestry University, Yangling 712100, China College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041, China
Zhuang He-Lin
Affiliation:
Laboratory of Animal Fat Deposition and Muscle Development, Northwest Agriculture and Forestry University, Yangling 712100, China
Yang Gong-She*
Affiliation:
Laboratory of Animal Fat Deposition and Muscle Development, Northwest Agriculture and Forestry University, Yangling 712100, China
*
Corresponding author: E-mail: gsyang999@yahoo.com.cn

Abstract

Using semi-quantitative reverse transcriptase polymerase chain reaction (semi-qRT-PCR), cell transfection, Hoechst 33342, acridine orange (AO) and flow cytometry methods, apoptotic changes in porcine pre-adipocytes treated with 9-cis retinoic acid (9-cisRA)–retinoid X receptor α (RXRα) ligand, transfected with pEnhanced green fluorescent proteins C2–RXRα (pEGFPC2–RXRα) or RXRα–small interfering RNA (RXRα–siRNA), respectively, were investigated. Observations of morphologic changes showed changes in cell structure, cellular shrinkage and condensation of chromatin, pyknosis of the nucleus into a compact body, complete fragmentation within the nuclear membrane, continuous budding and shedding, and, finally conversion of the cell into several apoptotic bodies wrapped in membranes of different sizes. Compared with the control group, the apoptotic rate was significantly lowered in the 10 nmol/l 9-cisRA-treated group, and significantly higher in the 10 μmol/l 9-cisRA-treated group (both P<0.05). Gene expression of RXRα was significantly up-regulated and down-regulated when cultured pre-adipocytes were transfected with pEGFPC2–RXRα or RXRα–siRNA (P<0.05), respectively; the apoptotic rate decreased in the pEGFPC2–RXRα group and significantly increased in the RXRα–siRNA group when compared with a control group (P<0.05). This implies that RXRα inhibits the apoptosis of porcine pre-adipocytes.

Type
Research Papers
Copyright
Copyright © China Agricultural University 2009

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Bellinger, DA, Merricks, EP and Nichols, TC (2006) Swine models of type 2 diabetes mellitus: insulin resistance, glucose tolerance, and cardiovascular complications. Institute for Laboratory Animal Research 47: 243258.CrossRefGoogle ScholarPubMed
Bonet, ML, Ribot, J, Felipe, F and Palou, A (2003) Vitamin A and the regulation of fat reserves. Cellular and Molecular Life Sciences 60: 13111321.CrossRefGoogle ScholarPubMed
Chawla, A, Schwarz, EJ, Dimaculangan, DD and Lazar, MA (1994) Peroxisome proliferator-activated receptor (PPAR) gamma: adipose-predominant expression and induction early in adipocyte differentiation. Endocrinology 135: 798800.CrossRefGoogle ScholarPubMed
Chen, LM, Chen, XC and Pan, XD (2008) Effects of IGFBP-3/RXRα or STAT-1 signaling pathway on Aβ1-42 induced apoptosis in rat hippocampus neurons. Acta Anatomica Sinica 39: 810815.Google Scholar
Fischer-Posovszky, P, Tornqvist, H, Debatin, KMI and Wabitsch, M (2004) Inhibition of death-receptor mediated apoptosis in human adipocytes by the insulin-like growth factor I (IGF-I)/IGF-I receptor autocrine circuit. Endocrinology 145: 18491859.CrossRefGoogle ScholarPubMed
Hellemans, K, Verbuyst, P, Quartier, E, et al. (2004) Differential modulation of rat hepatic stellate phenotype by natural and synthetic retinoids. Hepatology 39: 97–108.CrossRefGoogle ScholarPubMed
Imai, T, Jiang, M, Chambon, P and Metzger, D (2001) Impaired adipogenesis and lipolysis in the mouse upon selective ablation of the retinoid X receptor α mediated by a tamoxifen-inducible chimeric Cre recombinase (Cre-ERT2) in adipocytes. Proceedings of the National Academy of Sciences of the USA 98: 224228.Google ScholarPubMed
Lee, KW, Ma, L, Yan, X, Liu, B, Zhang, XK and Cohen, P (2005) Rapid apoptosis induction by IGFBP-3 involves an insulin-like growth factor-independent nucleomitochondrial translocation of RXR/Nur77. Journal of Biological Chemistry 280: 16942–16848.CrossRefGoogle Scholar
Levin, AA, Sturzenbecker, LJ, Kazmer, S, et al. (1992) 9-cis retinoic acid stereoisomer binds and activates the nuclear receptor RXRα. Nature 355: 359361.CrossRefGoogle Scholar
Li, Y, Yang, GS, Lu, RH and Sun, SD (2005) Optimal culture method of the porcine preadipocyte. Chinese Journal of Cell Biology 27: 697700 (in Chinese).Google Scholar
Lin, YQ, Li, RW, Sun, C, et al. (2004a) Rat adipocyte apoptosis induced by TNF-α. Journal of Agricultural Biotechnology 12: 644647 (in Chinese with English abstract).Google Scholar
Lin, XF, Zhao, BX, Chen, HZ, et al. (2004b) RXRα acts as a carrier for TR3 nuclear export in a 9-cis retinoic acid-dependent manner in gastric cancer cells. Journal of Cell Science 117: 56095621.CrossRefGoogle Scholar
Lin, YQ, Zhuang, HL and Yang, GS (2007a) The role of 9-cis RA in the regulation of pig preadipocyte differentiation. Journal of Molecular Cell Biology 40: 221226 (in Chinese).Google Scholar
Lin, YQ, Zhuang, HL and Yang, GS (2007b) Effects of RXRα gene silencing on the porcine adipocyte differentiation in vitro. Comparative Biochemistry and Physiology D 2: 207214.Google ScholarPubMed
Miller, WHJ, Faust, IM, Goldberger, AC and Hirsch, J (1983) Effects of severe long term food deprivation and refeeding on adipose tissue cells in the rat. American Journal of Physiology — Endocrinology and Metabolism 245: E74E80.CrossRefGoogle ScholarPubMed
Nagy, L, Thomazy, VA, Shipley, GL, et al. (1995) Activation of retinoid X receptors induces apoptosis in HL-60 cell lines. Molecular and Cellular Biology 15: 35403551.CrossRefGoogle ScholarPubMed
Prins, JB, Niesler, CU, Winterford, CM, et al. (1997) Tumor necrosis factor-alpha induces apoptosis of human adipose cells. Diabetes 46: 19391944.CrossRefGoogle ScholarPubMed
Prins, JB, Walker, NI, Winterford, CM and Cameron, DP (1994) Apoptosis of human adipocytes in vitro. Biochemical and Biophysical Research Communications 201: 500507.CrossRefGoogle ScholarPubMed
Reusch, JEB and Klemm, DJ (2002) Inhibition of cAMP-response element-binding protein activity decreases protein kinase B/Akt expression in 3T3-L1 adipocytes and induces apoptosis. The Journal of Biological Chemistry 277: 14261432.CrossRefGoogle ScholarPubMed
Szanto, A, Narkar, V, Shen, Q, Uray, IP, Davies, PJA and Nagy, L (2004) Retinoid X receptors: X-ploring their (patho)physiological functions. Cell Death and Differentiation 11: S126S143.CrossRefGoogle ScholarPubMed
Tontonoz, P, Graves, RA, Budavari, AI, et al. (1994) Adipocyte-specific transcription factor ARF6 is a heterodimeric complex of two nuclear hormone receptors, PPAR gamma and RXRα. Nucleic Acids Research 22: 56285634.CrossRefGoogle Scholar
Valladares, A, Alvarez, AM, Ventura, JJ, Roncero, C, Benito, M and Porras, A (2000) p38 mitogen-activated protein kinase mediates tumor necrosis factor-α-induced apoptosis in rat fetal brown adipocytes. Endocrinology 141: 43834395.CrossRefGoogle ScholarPubMed
Villarroya, F, Iglesias, R and Giralt, M (2004) Retinoids and retinoid receptors in the control of energy balance: novel pharmacological strategies in obesity and diabetes. Current Medicinal Chemistry 11: 795805.CrossRefGoogle ScholarPubMed
Wente, W, Brenner, MB, Zitzer, H, Gromada, J and Efanov, AM (2007) Activation of liver X receptors and retinoid X receptors induces growth arrest and apoptosis in insulin-secreting cells. Endocrinology 148: 18431849.CrossRefGoogle ScholarPubMed
Xiang, Y, Yao, D, Huang, SG and Wan, HY (2007) Synergistic effects of 9-cis-retinoic acid and 8-cl-cAMP on apoptosis of lung cancer cells. Journal of Shanghai Jiaotong University (Medical Science) 27: 405410– (in Chinese).Google Scholar
Zhang, XK (2007) Targeting Nur77 translocation. Expert Opinion on Therapeutic Targets 11: 6979.CrossRefGoogle ScholarPubMed
Zhao, Q, Shi, YR, Wei, XY, Wang, JF and Zhang, JH (2007) Role of 9-cis retinoic acid plus cytotoxic drugs in the proliferation and apoptosis of SH-SY5Y cell. Chinese Journal of Clinical Oncology 34: 12061208 (in Chinese).Google Scholar
Zhong, C, Yang, S, Huang, J, Cohen, MB and Roy-Burman, P (2003) Aberration in the expression of the retinoid receptor, RXRα, in prostate cancer. Cancer Biology and Therapy 2: 179184.CrossRefGoogle ScholarPubMed