Skip to main content
SpringerLink
Log in

Non-Esterified Fatty Acids Activate the AMP-Activated Protein Kinase Signaling Pathway to Regulate Lipid Metabolism in Bovine Hepatocytes

  • Original Paper
  • Published:
Cell Biochemistry and Biophysics Aims and scope Submit manuscript

Abstract

Non-esterified fatty acids (NEFAs) act as signaling molecules involved in regulating genes expression to modulate lipid metabolism. However, the regulation mechanism of NEFAs on lipid metabolism in dairy cows is unclear. The AMP-activated protein kinase (AMPK) signaling pathway plays a key role in regulating hepatic lipid metabolism. In vitro, bovine hepatocytes were cultured and treated with different concentrations of NEFAs and AMPKα inhibitors (BML-275). NEFAs increased AMPKα phosphorylation through up-regulating the protein levels of liver kinase B1. Activated AMPKα increased the expression and transcriptional activity of peroxisome proliferator-activated receptor α (PPARα). NEFAs also directly activate the PPARα independent of AMPKα. Activated PPARα increased the lipolytic genes expression to increase lipid oxidation. Furthermore, activated AMPKα inhibited the expression and transcriptional activity of the sterol regulatory element-binding protein 1c and carbohydrate responsive element-binding protein, which reduced the expression of lipogenic genes, thereby decreasing lipid synthesis. Activated AMPKα phosphorylated and inhibited acetyl-CoA carboxylase and increased carnitine palmitoyltransferase-1 activity, which increased lipid oxidation. Consequently, the triglyceride content in the NEFAs-treated hepatocytes was significantly decreased. These results indicate that NEFAs activate the AMPKα signaling pathway to increase lipid oxidation and decrease lipid synthesis in hepatocytes, which in turn, generates more ATP to relieve the negative energy balance in transition dairy cows.

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

NEB:

Negative energy balance

NEFAs:

Non-esterified fatty acids

AMPK:

AMP-activated protein kinase

LKB1:

Liver kinase B1

PPARα:

Peroxisome proliferator-activated receptor α

SREBP-1c:

Sterol regulatory element-binding protein 1c

ChREBP:

Carbohydrate responsive element-binding protein

TG:

Triglyceride

ACO:

Acyl-CoA oxidase

CPT1:

Carnitine palmitoyltransferase 1

CPT2:

Carnitine palmitoyltransferase 2

L-FABP:

Liver fatty acid-binding protein

ACSL:

Acyl-CoA synthetase long-chain

ACC1:

Acetyl-CoA carboxylase 1

p-ACC1:

Phosphorylated-ACC1

FAS:

Fatty acid synthase

SCD-1:

Stearoyl-CoA desaturase-1

p-AMPKα:

Phosphorylated-AMPKα

EMSA:

Electrophoretic mobility shift assay

PUFAs:

Polyunsaturated fatty acids

References

  1. Dann, H. M., & Drackley, J. K. (2006). Carnitine palmitoyltransferase I in liver of periparturient dairy cows: effects of prepartum intake, postpartum induction of ketosis, and periparturient disorders. Journal of Dairy Science, 88, 3851–3859.

    Article  Google Scholar 

  2. Loor, J. J., Everts, R. E., Bionaz, M. H., Dann, M., Morin, D. E., Oliveira, R., et al. (2007). Nutrition-induced ketosis alters metabolic and signaling gene networks in liver of periparturient dairy cows. Physiological Genomics, 32, 105–116.

    Article  PubMed  CAS  Google Scholar 

  3. Wathes, D. C., Cheng, Z., Fenwick, M. A., Fitzpatrick, R., & Patton, J. (2011). Influence of energy balance on the somatotrophic axis and matrix metalloproteinase expression in the endometrium of the postpartum dairy cow. Reproduction, 141, 269–281.

    Article  PubMed  CAS  Google Scholar 

  4. Xu, C., Wang, Z., Liu, G. W., Li, X. B., Xie, G. H., Xia, C., et al. (2008). Metabolic characteristic of the liver of dairy cows during ketosis based on comparative proteomics. Asian-Australian Journal of Animal Sciences, 21, 1003–1010.

    CAS  Google Scholar 

  5. Jorritsma, R., Jorritsma, H., Schukken, Y. H., Bartlettd, P. C., Wensinga, T., & Wentink, G. H. (2001). Prevalence and indicators of post partum fatty infiltration of the liver in nine commercial dairy herds in The Netherlands. Livestock Production Science, 68, 53–60.

    Article  Google Scholar 

  6. Zhang, Z., Li, X., Liu, G., Gao, L., Guo, C., Kong, T., et al. (2011). High insulin concentrations repress insulin receptor gene expression in calf hepatocytes cultured in vitro. Cellular Physiology and Biochemistry, 27, 637–640.

    Article  PubMed  CAS  Google Scholar 

  7. Duplus, E., Glorian, M., & Forest, C. (2000). Fatty acid regulation of gene transcription. Journal of Biological Chemistry, 275, 30749–30752.

    Article  PubMed  CAS  Google Scholar 

  8. Jump, D. B., Botolin, D., Wang, Y., Xu, J., Christian, B., & Demeure, O. (2005). Fatty acid regulation of hepatic gene transcription. Journal of Nutrition, 135, 2503–2506.

    PubMed  CAS  Google Scholar 

  9. Kahn, B. B., Alquier, T., Carling, D., & Hardie, D. G. (2005). AMP-activated protein kinase: ancient energy gauge provides clues to modern understanding of metabolism. Cell Metabolism, 1, 15–25.

    Article  PubMed  CAS  Google Scholar 

  10. Hardie, D. G. (2003). The AMP-activated protein kinase cascade: the key sensor of cellular energy status. Endocrinology, 144, 5179–5183.

    Article  PubMed  CAS  Google Scholar 

  11. Viollet, B., Foretz, M., Guigas, B., Horman, S., Dentin, R., Bertrand, L., et al. (2006). Activation of AMP-activated protein kinase in the liver: A new strategy for the management of metabolic hepatic disorders. Journal of Physiology, 574(Pt 1), 41–53.

    Article  PubMed  CAS  Google Scholar 

  12. Long, Y. C., & Zierath, J. R. (2006). AMP-activated protein kinase signaling in metabolic regulation. The Journal of Clinical Investigation, 116, 1776–1783.

    Article  PubMed  CAS  Google Scholar 

  13. Martínez, N., White, V., Kurtz, M., Higa, R., Capobianco, E., & Jawerbaum, A. (2011). Activation of the nuclear receptor PPARα regulates lipid metabolism in foetal liver from diabetic rats: Implications in diabetes-induced foetal overgrowth. Diabetes/Metabolism Research and Reviews, 27, 35–46.

    Article  PubMed  Google Scholar 

  14. Uyeda, K., & Repa, J. J. (2006). Carbohydrate response element binding protein, ChREBP, a transcription factor coupling hepatic glucose utilization and lipid synthesis. Cell Metabolism, 4, 107–110.

    Article  PubMed  CAS  Google Scholar 

  15. Kawaguchi, T., Osatomi, K., Yamashita, H., Kabashima, T., & Uyeda, K. (2002). Mechanism for fatty acid “sparing” effect on glucose-induced transcription: Regulation of carbohydrate-responsive element-binding protein by AMP-activated protein kinase. Journal of Biological Chemistry, 277, 3829–3835.

    Article  PubMed  CAS  Google Scholar 

  16. Li, Y., Xu, S., Mihaylova, M. M., Zheng, B., Hou, X., Jiang, B., et al. (2011). AMPK phosphorylates and inhibits SREBP activity to attenuate hepatic steatosis and atherosclerosis in diet-induced insulin-resistant mice. Cell Metabolism, 13, 376–388.

    Article  PubMed  CAS  Google Scholar 

  17. Shaw, R. J., Lamia, K. A., Vasquez, D., Koo, S. H., Bardeesy, N., Depinho, R. A., et al. (2005). The kinase LKB1 mediates glucose homeostasis in liver and therapeutic effects of metformin. Science, 310, 1642–1646.

    Article  PubMed  CAS  Google Scholar 

  18. Xu, C., Wang, Z., Zhang, R. H., Zhang, H. Y., Fu, S. X., & Xia, C. (2011). Effect of NEFA and glucose levels on CPT-I mRNA expression and translation in cultured bovine hepatocytes. Journal of Veterinary Medical Science, 73, 97–101.

    Article  PubMed  CAS  Google Scholar 

  19. Zhang, Z. G., Li, X. B., Gao, L., Liu, G. W., Kong, T., Li, Y. F., et al. (2012). An updated method for the isolation and culture of primary calf hepatocytes. Veterinary Journal, 191, 323–326.

    Article  Google Scholar 

  20. Li, X., Li, X., Bai, G., Chen, H., Deng, Q., Liu, Z., et al. (2012). Effects of non-esterified fatty acids on the gluconeogenesis in bovine hepatocytes. Molecular and Cellular Biochemistry, 359, 385–388.

    Article  PubMed  CAS  Google Scholar 

  21. Ramnanan, C. J., McMullen, D. C., Groom, A. G., & Storey, K. B. (2010). The regulation of AMPK signaling in a natural state of profound metabolic rate depression. Molecular and Cellular Biochemistry, 335, 91–105.

    Article  PubMed  CAS  Google Scholar 

  22. Yu, P. B., Hong, C. C., Sachidanandan, C., Babitt, J. L., Deng, D. Y., Hoyng, S. A., et al. (2008). Dorsomorphin inhibits BMP signals required for embryogenesis and iron metabolism. Nature Chemical Biology, 4, 33–41.

    Article  PubMed  CAS  Google Scholar 

  23. Cameron, R. E., Dyk, P. B., Herdt, T. H., Kaneene, J. B., Miller, R., Bucholtz, H. F., et al. (1998). Dry cow diet, management, and energy balance as risk factors for displaced abomasums in high producing dairy herds. Journal of Dairy Science, 81, 132–139.

    Article  PubMed  CAS  Google Scholar 

  24. Li, P., Li, X. B., Fu, S. X., Wu, C. C., Wang, X. X., Yu, G. J., et al. (2012). Alterations of fatty acid β-oxidation capability in the liver of ketotic cows. Journal of Dairy Science, 95, 1759–1766.

    Article  PubMed  CAS  Google Scholar 

  25. Andreelli, F., Foretz, M., Knauf, C., Cani, P. D., Perrin, C., Iglesias, M. A., et al. (2006). Liver adenosine monophosphate-activated kinase-alpha 2 catalytic subunit is a key target for the control of hepatic glucose production by adiponectin and leptin but not insulin. Endocrinology, 147, 2432–2441.

    Article  PubMed  CAS  Google Scholar 

  26. Foretz, M., Ancellin, N., Andreelli, F., Saintillan, Y., Grondin, P., Kahn, A., et al. (2005). Short-term overexpression of a constitutively active form of AMP-activated protein kinase in the liver leads to mild hypoglycemia and fatty liver. Diabetes, 54, 1331–1339.

    Article  PubMed  CAS  Google Scholar 

  27. Suchankova, G., Tekle, M., Saha, A., Ruderman, N. B., Clarke, S. D., & Gettys, T. W. (2005). Dietary polyunsaturated fatty acids enhance hepatic AMP-activated protein kinase activity in rats. Biochemical and Biophysical Research Communications, 326, 851–858.

    Article  PubMed  CAS  Google Scholar 

  28. Clark, H., Carling, D., & Saggerson, D. (2004). Covalent activation of heart AMP-activated protein kinase in response to physiological concentrations of long-chain fatty acids. European Journal of Biochemistry, 271, 2215–2224.

    Article  PubMed  CAS  Google Scholar 

  29. Fediuc, S., Gaidhu, M. P., & Ceddia, R. B. (2006). Regulation of AMP-activated protein kinase and acetyl-CoA carboxylase phosphorylation by palmitate in skeletal muscle cells. Journal of Lipid Research, 47, 412–420.

    Article  PubMed  CAS  Google Scholar 

  30. Watt, M. J., Steinberg, G. R., Chen, Z. P., Kemp, B. E., & Febbraio, M. A. (2006). Fatty acids stimulate AMP activated protein kinase and enhance fatty acid oxidation in L6 myotubes. The Journal of Physiology, 574, 139–147.

    Article  PubMed  CAS  Google Scholar 

  31. Zhou, G., Myers, R., Li, Y., Chen, Y., Shen, X., Fenyk-Melody, J., et al. (2001). Role of AMP-activated protein kinase in mechanism of metformin action. The Journal of Clinical Investigation, 108, 1167–1174.

    PubMed  CAS  Google Scholar 

  32. Qiu, L., Wu, X., Chau, J. F., Szeto, I. Y., Tam, W. Y., Guo, Z., et al. (2008). Aldose reductase regulates hepatic peroxisome proliferator-activated receptor alpha phosphorylation and activity to impact lipid homeostasis. Journal of Biological Chemistry, 283, 17175–17183.

    Article  PubMed  CAS  Google Scholar 

  33. Sugden, M. C., Bulmer, K., Gibbons, G. F., Knight, B. L., & Holness, M. J. (2002). Peroxisome proliferator-activated receptor-a (PPARa) deficiency leads to dysregulation of hepatic lipid and carbohydrate metabolism by fatty acids and insulin. Biochemical Journal, 364, 361–368.

    Article  PubMed  CAS  Google Scholar 

  34. Everett, L., Galli, A., & Crabb, D. (2000). The role of hepatic peroxisome proliferator-activated receptors (PPARs) in health and disease. Liver, 20, 191–199.

    Article  PubMed  CAS  Google Scholar 

  35. Pawar, A., & Jump, D. B. (2003). Unsaturated fatty acid regulation of PPARα activity in primary rat hepatocytes. Journal of Biological Chemistry, 278, 35931–35939.

    Article  PubMed  CAS  Google Scholar 

  36. Steven, D. C. (2001). Polyunsaturated fatty acid regulation of gene transcription: A molecular mechanism to improve. American Society for Nutritional Sciences, 60, 1129–1132.

    Google Scholar 

  37. Shimomura, I., Bashmakov, Y., & Horton, J. D. (1999). Increased levels of nuclear SREBP-1c associated with fatty livers in two mouse models of diabetes mellitus. Journal of Biological Chemistry, 274, 30028–30032.

    Article  PubMed  CAS  Google Scholar 

  38. Iizuka, K., Bruick, R. K., Liang, G., Horton, J. D., & Uyeda, K. (2001). Deficiency of carbohydrate response element-binding protein (ChREBP) reduces lipogenesis as well as glycolysis. Proceedings of the National Academy of Sciences of the United States of America, 101, 7281–7286.

    Article  Google Scholar 

  39. Dobrzyn, A., Dobrzyn, P., Miyazaki, M., & Ntambi, J. M. (2005). Polyunsaturated fatty acids do not activate AMP-activated protein kinase in mouse tissues. Biochemical and Biophysical Research Communications, 332, 892–896.

    Article  PubMed  CAS  Google Scholar 

  40. Dentin, R., Benhamed, F., Pégorier, J. P., Foufelle, F., Viollet, B., Vaulont, S., et al. (2005). Polyunsaturated fatty acids suppress glycolytic and lipogenic genes through the inhibition of ChREBP nuclear protein translocation. The Journal of Clinical Investigation, 115, 2843–2854.

    Article  PubMed  CAS  Google Scholar 

  41. Hardie, D. G. (2004). The AMP-activated protein kinase pathway-new players upstream and downstream. Journal of Cell Science, 117, 5479–5487.

    Article  PubMed  CAS  Google Scholar 

  42. Ruderman, N. B., Saha, A. K., Vavvas, D., & Witters, L. A. (1999). Malonyl-CoA fuel sensing, and insulin resistance. American Journal of Physiology, Endocrinology and Metabolism, 276, 1–18.

    Google Scholar 

Download references

Acknowledgments

This study was supported by the National Key Basic Research Program of China (No. 2011CB100800).

Author information

Authors and Affiliations

  1. Key Laboratory of Zoonosis, Ministry of Education, College of Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun, 130062, Jilin, China

    Xinwei Li, Xiaobing Li, Hui Chen, Liancheng Lei, Juxiong Liu, Yuan Guan, Zhaoxi Liu, Liang Zhang, Wentao Yang, Chenxu Zhao, Guowen Liu & Zhe Wang

  2. Institute of Animal Science and Technology, Heilongjiang Bayi Agriculture University, Daqing, 163319, Heilongjiang, China

    Shixin Fu

  3. College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang, 110161, Liaoning, China

    Peng Li

Authors
  1. Xinwei Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiaobing Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hui Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Liancheng Lei
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Juxiong Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yuan Guan
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Zhaoxi Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Liang Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Wentao Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Chenxu Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Shixin Fu
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Peng Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Guowen Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Zhe Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Guowen Liu or Zhe Wang.

Additional information

Xinwei Li and Xiaobing Li contributed equally to this study.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Li, X., Li, X., Chen, H. et al. Non-Esterified Fatty Acids Activate the AMP-Activated Protein Kinase Signaling Pathway to Regulate Lipid Metabolism in Bovine Hepatocytes. Cell Biochem Biophys 67, 1157–1169 (2013). https://doi.org/10.1007/s12013-013-9629-1

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12013-013-9629-1

Keywords

Search

Navigation