Skip to main content

Advertisement

SpringerLink
Log in

Establishment of the Insulin Resistance Induced by Inflammatory Response in 3T3-L1 Preadipocytes Cell Line

  • Published:
Inflammation Aims and scope Submit manuscript

Abstract

In the light of given recent reports, insulin resistance related to inflammation is characterized by increasing a diverse array of pro-inflammatory cytokines. In this study, hypothesizing that 3T3-L1 non-differentiated preadipocytes cell line as a cell model could be used to investigate this linkage, the aim is to determine whether the preadipocytes induced by different inflammatory responses could cause insulin resistance. This paper has determined the time and concentration-dependent effects of insulin on glucose consumption in the 3T3-L1 non-differentiated preadipocytes. Glucose consumption has also been assayed in the preadipocytes which are treated with LPS and CM originated from LPS-activated RAW264.7. Then protein level of each group has been measured by coomassie brilliant blue protein kit. Furthermore, secretion levels of IL-6 and TNF-α are measured by ELISA in the supernatant of RAW264.7 and preadipocytes. Finally, the mRNA expressions for IL-6, TNF-α and PPARγ has been assessed by RT-PCR. The results show that administration of LPS and CM both can increase releases of IL-6 and TNF-α, as well as gene expression of IL-6 mRNA; this change is accompanied with suppression of PPARγ mRNA activation in 3T3-L1 undifferentiated preadipocytes. In conclusion, our results suggest that in preadipocytes, pro-inflammatory cytokines can result in insulin resistance, and deserve further investigation to be helpful for treatment and revealing mechanisms of T2DM.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

Abbreviations

LPS:

lipopolysaccharide

RT-PCR:

reverse transcription polymerase chain reaction

ELISA:

enzyme- linked immunosorbent assay

TNF-α:

tumor necrosis factor-alpha

IL-6:

interleukin-6

PPARγ:

peroxisome proliferators activated receptor-gamma

CM:

conditioned media of LPS-activated RAW264.7

T2DM:

type 2 diabetes mellitus

GOD-POD:

glucose oxidase-peroxidase

PBS:

phosphate buffered solution

EDTA:

Ethylenediaminetetraaceticacid

SDS:

sodium dodecyl sulfate

Tris–HCl:

Tris (hydroxymethyl) aminomethane hydrochloride

References

  1. DeFronzo, R. A. 1997. Insulin resistance: a multifaceted syndrome responsible for NIDDM, obesity, hypertension, yslipidaemia and atherosclerosis. Neth. J. Med. 50:191–197. doi:10.1016/S0300-2977(97)00012-0.

    Article  PubMed  CAS  Google Scholar 

  2. Weyer, C., C. Bogardus, D. M. Mott, and R. E. Pratley. 1999. The natural history of insulin secretory dysfunction and insulin resistance in the pathogenesis of type 2 diabetes mellitus. J. Clin. Invest. 104:787–794. doi:10.1172/JCI7231.

    Article  PubMed  CAS  Google Scholar 

  3. Coppack, S. W. 2001. Pro-inflammatory cytokines and adipose tissue. Proc. Nutr. Soc. 60:349–356.

    Article  PubMed  CAS  Google Scholar 

  4. Hotamisligil, G. S., N. S. Shargill, and B. M. Spiegelman. 1993. Adipose expression of tumor necrosis factor-α: direct role in obesity-linked insulin resistance. Science 259:87–91. doi:10.1126/science.7678183.

    Article  PubMed  CAS  Google Scholar 

  5. Feinstein, R., H. Kanety, M. Z. Papa, B. Lunenfeld, and A. Karasik. 1993. Tumor necrosis factor-α suppresses insulin-induced tyrosine phosphorylation of insulin receptor and its substrates. J. Biol. Chem. 268:26055–26058.

    PubMed  CAS  Google Scholar 

  6. Bastard, J. P., M. Maachi, J. T. Van Nhieu, C. Jardel, E. Bruckert, A. Grimaldi et al. 2002. Adipose tissue IL-6 content correlates with resistance to insulin activation of glucose uptake both in vivo and in vitro. J. Clin. Endocrinol. Metab. 87:2084–2089. doi:10.1210/jc.87.5.2084.

    Article  PubMed  CAS  Google Scholar 

  7. Barbara, B. K., and S. F. Jeffrey. 2000. Obesity and insulin resistance. J. Clin. Invest. 106:473–481. doi:10.1172/JCI10842.

    Article  Google Scholar 

  8. Wellen, K. E., and G. S. Hotamisligil. 2003. Obesity-induced inflammatory changes in adipose tissue. J. Clin. Invest. 112(12):1785–1788.

    PubMed  CAS  Google Scholar 

  9. Marià, A., A. José, and L. Fernández. 2006. Adipose tissue: something more than just adipocytes. Curr. Nutr. Food Sci. 2:141–150. doi:10.2174/157340106776818817.

    Article  Google Scholar 

  10. Odile, P. G., and F. Philippe. 2007. Preadipocyte response and impairment of differentiation in an inflammatory environment. Biochem. Biophys. Res. Commun. 356:662–667. doi:10.1016/j.bbrc.2007.03.053.

    Article  CAS  Google Scholar 

  11. Cousin, B., O. Munoz, and M. Andre. 1999. A role for preadipocytes as macrophage-like cells. FASEB J. 13:305–312.

    PubMed  CAS  Google Scholar 

  12. Harkins, J. M., M. N. Moustaid, Y. Chung, K. M. Penner, J. Pestka, C. M. North et al. 2004. Expression of interleukin-6 is greater in preadipocytes than in adipocytes of 3T3-L1 cells and C57BL/6J and ob/ob mice. J. Nutr. 134:2673–2677.

    PubMed  CAS  Google Scholar 

  13. Ken, Y., K. Daisuke, O. Yasushi, and K. Koichiro. 2004. A novel preadipocyte cell line established from mouse adult mature adipocytes. Biochem. Biophys. Res. Commun. 321:967–974. doi:10.1016/j.bbrc.2004.07.055.

    Article  CAS  Google Scholar 

  14. Wen, G., W. Siu, W. S. Xie, T. L. Lei, and Z. J. Luo. 2007. Palmitate modulates intracellular signaling, induces endoplasmic reticulum stress, and causes apoptosis in mouse 3T3-L1 and rat primary preadipocytes. Am. J. Physiol. Endocrinol. Metab. 293:576–586. doi:10.1152/ajpendo.00523.2006.

    Article  CAS  Google Scholar 

  15. Suganami, T., M. Mukoyama, A. Sugawara, K. Mori, T. Nagae, M. Kasahara et al. 2001. Over-expression of brain natriuretic peptide in mice ameliorates immune-mediated renal injury. J. Am. Soc. Nephrol. 12:2652–2663.

    PubMed  CAS  Google Scholar 

  16. Yamamoto, Y., Y. Yoshimasa, M. Koh, J. Suga, H. Masuzaki, Y. Ogawa et al. 2000. Constitutively active mitogen-activated protein kinase kinase increases GLUT1 expression and recruits both GLUT1 and GLUT4 at the cell surface in 3T3-L1 adipocytes. Diabetes 49:332–339. doi:10.2337/diabetes.49.3.332.

    Article  PubMed  CAS  Google Scholar 

  17. Kentaro, M., B. Hideaki, U. Hiroyuki, and S. Yasushi. 2007. Effect of PPARα activation of macrophages on the secretion of inflammatory cytokines in cultured adipocytes. Eur. J. Pharmacol. 561:206–213. doi:10.1016/j.ejphar.2006.12.037.

    Article  CAS  Google Scholar 

  18. Paska, A. P., M. Christopher, and D. R. Peter. 2006. Macrophage-secreted factors induce adipocyte inflammation and insulin resistance. Biochem. Biophys. Res. Commun. 341:507–514. doi:10.1016/j.bbrc.2006.01.012.

    Article  CAS  Google Scholar 

  19. Victoria, R., N. Ivan, and S. Ulf. 2003. Interleukin-6 (IL-6) induces insulin resistance in 3T3-L1 adipocytes and Is, Like IL-8 and tumor necrosis Factor-α, over-expressed in human fat cells from insulin-resistant subjects. J. Biol. Chem. 278:45777–45784. doi:10.1074/jbc.M308352200.

    Article  Google Scholar 

  20. Esposito, K., A. Pontillo, P. C. Di, G. Giugliano, M. Masella, R. Marfella et al. 2003. Effect of weight loss and lifestyle changes on vascular inflammatory markers in obese women: a randomized trial. JAMA 289:1799–1804. doi:10.1001/jama.289.14.1799.

    Article  PubMed  CAS  Google Scholar 

  21. Bruun, J. M., C. Verdich, S. Toubro, A. Astrup, and B. Richelsen. 2003. Association between measures of insulin sensitivity and circulating levels of interleukin-8, interleukin-6, and tumor necrosis factor-alpha. Effect of weight loss in obese men. Eur. J. Endocrinol. 148:535–542. doi:10.1530/eje.0.1480535.

    Article  PubMed  CAS  Google Scholar 

  22. Mohamed-Ali, V., S. Goodrick, A. Rawesh, D. R. Katz, J. M. Miles, J. S Yudkin et al. 1997. Subcutaneous adipose tissue releases interleukin-6, but not tumor necrosis factor a, in vivo. J. Clin. Endocrinol. Metab. 82:4196–4200. doi:10.1210/jc.82.12.4196.

    Article  PubMed  CAS  Google Scholar 

  23. Fried, S. K., D. A. Bunkin, and A. S. Greenberg. 2003. Omental and subcutaneous adipose tissues of obese subjects release interleukin-6: depot difference and regulation by glucocorticoid. J. Clin. Endocrinol. Metab. 83:847–850. doi:10.1210/jc.83.3.847.

    Article  Google Scholar 

  24. Hotamisligil, G. S. 2003. Inflammatory pathways and insulin action. Int. J. Obes. Relat. Metab. Disord. 27(s3):53–55. doi:10.1038/sj.ijo.0802502.

    Article  CAS  Google Scholar 

  25. Lyon, C. J., R. E. Law, and W. A. Hsueh. 2003. Mini-review: adiposity, inflammation, and atherogenesis. Endorinol. 144:2195–2200. doi:10.1210/en.2003-0285.

    Article  CAS  Google Scholar 

  26. Yamini, B. T., T. Pratibha, K. Kiranmayi, C. C. Sheau, J. S. Brenda, and H. A. Bahram. 2008. Role of Sandhika: a polyherbal formulation on MC3T3-E1 osteoblast-like cells. Inflammation 31:1–8. doi:10.1007/s10753-007-9044-0.

    Article  Google Scholar 

  27. Fain, J. N., S. W. Bahouth, and A. K. Madan. 2004. TNF-alpha release by the nonfat cells of human adipose tissue. Int. J. Obes. Relat. Metab. Disord. 28:616–622. doi:10.1038/sj.ijo.0802594.

    Article  PubMed  CAS  Google Scholar 

  28. Matthias, H., N. E. Alex, P. Ralph, P. Thomas, S. Dalma, K. Ulrich et al. 2008. LPS induces interleukin-6 and interleukin-8 but not tumor necrosis factor-α in human adipocytes. Cytokine 41:29–37. doi:10.1016/j.cyto.2007.10.008.

    Article  CAS  Google Scholar 

Download references

Acknowledgment

This research was financially supported by the Jiangsu Science Foundation of China (BK2007172). The authors also thank the anonymous referees for the comments and checking the language of the manuscript.

Author information

Authors and Affiliations

  1. College of Pharmacology of Chinese Materia Medica, China Pharmaceutical University, Nanjing, 210038, People’s Republic of China

    Yuan Zhou, Bao-lin Liu, Kang Liu, Yuan An & Lin Li

  2. Quality Test Centrol of Animal Product in Jiangsu, Nanjing, 210036, People’s Republic of China

    Yuan Zhou & Jue Huang

  3. Life Science Department, Nanjing Xiaozhuang College, Nanjing, 211171, People’s Republic of China

    Ning Tang

Authors
  1. Yuan Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bao-lin Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kang Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ning Tang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jue Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yuan An
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Lin Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Bao-lin Liu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhou, Y., Liu, Bl., Liu, K. et al. Establishment of the Insulin Resistance Induced by Inflammatory Response in 3T3-L1 Preadipocytes Cell Line. Inflammation 31, 355–364 (2008). https://doi.org/10.1007/s10753-008-9086-y

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10753-008-9086-y

KEY WORDS

Search

Navigation