Abstract
Pancreatic β-cell dysfunction is an important pathological change in type 2 diabetes, which is tightly related to obesity. However, the direct role of adipose tissue in β-cell dysfunction has not been well understood. In this study, we examined the effects of 3T3-L1 adipocytes on MIN6 insulin-secreting cells in a co-culture system. MIN6 cells used here kept most of β-cell functions but less sensitive to glucose stimulation. Tolbutamide, the KATP channel blocker, was therefore used to stimulate insulin secretion in this report. MIN6 cells co-cultured with 3T3-L1 adipocytes had significantly reduced intracellular calcium concentration ([Ca2+]i) and lost the ability to secrete insulin in response to tolbutamide, compared to the control cells. 3T3-L1 adipocytes significantly decreased the expression of insulin, glucokinase and Kir6.2 genes but increased the expression of uncoupling protein-2 (UCP-2) in MIN6 cells after one week of co-culture, as measured by semi-quantitative RT-PCR. 3T3-L1 adipocyte-conditioned medium also significantly decreased insulin secretion and the expression of insulin, glucokinase and Kir6.2 genes in MIN6 cells. The conditioned medium also reduced tyrosine kinase activity in MIN6 cells. The inhibitor of protein tyrosine kinase, genistein, decreased the expression of glucokinase and Kir6.2 in MIN6 cells, while two free fatty acids, oleic acid and linoleic acids, were found to increase UCP-2 expression. The present study demonstrates that 3T3-L1 adipocytes directly impair insulin secretion and the␣expression of important genes in MIN6 cells. The effects of␣3T3-L1 adipocytes on MIN6 cells are ascribed to␣secreted bioactive factors and may be mediated via multiple pathways, which include the upregulation of UCP-2 expression via free fatty acids, and downregulation of glucokinase and Kir6.2 expression via decreasing protein tyrosine kinase activity.
Similar content being viewed by others
References
S.E. Kahn, Am. J. Med. 108, 2S (2000)
V. Poitout, R.P. Robertson, Annu. Rev. Med. 47, 69 (1996)
Y. Tokuyama, J. Sturis, A.M. DePaoli et al., Diabetes 44, 1447 (1995)
H. Yki-Jarvinen, Endocr. Rev. 13, 415 (1992)
R.H. Unger, Diabetes 44, 863 (1995)
Y. Lee, H. Hirose, M. Ohneda, J.H. Johnson, J.D. McGarry, R.H. Unger, Proc. Natl. Acad. Sci. USA 91, 10878 (1994)
I. Briaud, J.S. Harmon, C.L. Kelpe, V.B. Segu, V. Poitout, Diabetes 50, 315 (2001)
V. Poitout, R.P. Robertson, Endocrinology 143, 339 (2002)
P.G. Kopelman, L. Albon, Br. Med. Bull. 53, 322 (1997)
A.J. Scheen, Acta Clin. Belg. 55, 9 (2000)
G. Fruhbeck, J. Gomez-Ambrosi, F.J. Muruzabal, M.A. Burrell, Am. J. Physiol. Endocrinol. Metab. 280, E827 (2001)
T.J. Kieffer, R.S. Heller, J.F. Habener, Biochem. Biophys. Res. Commun. 224, 522 (1996)
I. Kharroubi, J. Rasschaert, D.L. Eizirik, M. Cnop, Biochem. Biophys. Res. Commun. 312, 1118 (2003)
C.P. Briscoe, M. Tadayyon, J.L. Andrews, et al., J. Biol. Chem. 278, 11303. Epub 12002 Dec 11319 (2003)
Y.F. Zhao, Y.L. Zhu, C. Chen, Sheng Li Xue Bao 56, 253 (2004)
A.P. Babenko, L. Aguilar-Bryan, J. Bryan, Annu. Rev. Physiol. 60, 667 (1998)
T. Miki, K. Nagashima, F. Tashiro et al., Proc. Natl. Acad. Sci. USA 95, 10402 (1998)
H. Huopio, S.L. Shyng, T. Otonkoski, C.G. Nichols, Am. J. Physiol. Endocrinol. Metab. 283, E207 (2002)
C. Postic, M. Shiota, M.A. Magnuson, Recent Prog. Horm. Res. 56, 195 (2001)
F.M. Matschinsky, Diabetes 39, 647 (1990)
A.T. Hattersley, P.M. Clark, R. Page et al., Diabetologia 40, 1367 (1997)
S.S. Fajans, G.I. Bell, D.W. Bowden, J.B. Halter, K.S. Polonsky, Life Sci. 55, 413 (1994)
C.Y. Zhang, G. Baffy, P. Perret et al., Cell 105, 745 (2001)
M.Y. Wang, M. Shimabukuro, Y. Lee et al., Diabetes 48, 1020 (1999)
C.B. Chan, P.E. MacDonald, M.C. Saleh, D.C. Johns, E. Marban, M.B. Wheeler, Diabetes 48, 1482 (1999)
Y.F. Zhao, D.J. Keating, M. Hernandez, D.D. Feng, Y. Zhu, C. Chen, J. Mol. Endocrinol. 35, 49 (2005)
H. Yoshikawa, Y. Tajiri, Y. Sako, T. Hashimoto, F. Umeda, H. Nawata, Metabolism 50, 613 (2001)
L.C. Bollheimer, R.H. Skelly, M.W. Chester, J.D. McGarry, C.J. Rhodes, J. Clin. Invest. 101, 1094 (1998)
R.N. Bergman, M. Ader, Trends Endocrinol. Metab. 11, 351 (2000)
L.X. Li, F. Skorpen, K. Egeberg, I.H. Jorgensen, V. Grill, Endocrinology 143, 1371 (2002)
P. Jezek, H. Engstova, M. Zackova et al., Biochim. Biophys. Acta 1365, 319 (1998)
N. Lameloise, P. Muzzin, M. Prentki, F. Assimacopoulos-Jeannet, Diabetes 50, 803 (2001)
Y. Itoh, Y. Kawamata, M. Harada et al., Nature 422, 173 (2003)
C.P. Briscoe, M. Tadayyon, J.L. Andrews et al., J. Biol. Chem. 278, 11303 (2003)
T.J. O’Dell, E.R. Kandel, S.G. Grant, Nature 353, 558 (1991)
S.M. Appleyard, J.P. McLaughlin, C. Chavkin, J. Biol. Chem. 275, 38281 (2000)
H. Ishihara, T. Asano, K. Tsukuda et al., Diabetologia 36, 1139 (1993)
V. Poitout, L.K. Olson, R.P. Robertson, Diabetes Metab. 22, 7 (1996)
K. Peppel, C. Baglioni, Biotechniques 9, 711 (1990)
M. Jackerott, A. Baudry, B. Lamothe, D. Bucchini, J. Jami, R.L. Joshi, Diabetes 50, S146 (2001)
Acknowledgements
We express our thanks to Dr M. Garry (Department of Biochemistry, Monash University, Melbourne, Australia) for supplying MIN6 cells, with the approval of Dr. J. Miyazaki (Osaka University, Osaka, Japan). This work was supported by the Australian NH&MRC, Prince Henry’s Institute of Medical Research, MAZDA Foundation, Eli Lilly Australia, and the Nature Science Foundation of China.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Zhao, YF., Feng, DD., Hernandez, M. et al. 3T3-L1 adipocytes induce dysfunction of MIN6 insulin-secreting cells via multiple pathways mediated by secretory factors in a co-culture system. Endocr 31, 52–60 (2007). https://doi.org/10.1007/s12020-007-0001-3
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12020-007-0001-3