Journal of Molecular Medicine

, Volume 94, Issue 2, pp 183–194 | Cite as

Inhibition of AMPK through Lyn-Syk-Akt enhances FcεRI signal pathways for allergic response

  • Kai-Chun Lin
  • Duen-Yi Huang
  • De-Wei Huang
  • Shiang-Jong Tzeng
  • Wan-Wan Lin
Original Article


AMPK was shown to negatively regulate FcεRI activation, and FcεR-mediated Fyn activation can counteract the LKB1/AMPK axis in mast cells. However, the relationship between the major Src family kinase Lyn and AMPK remains poorly defined. Here, we investigate the molecular mechanism for AMPK inhibition by FcεRI-Lyn signaling in rat RBL-2H3 cells. We found that FcεRI activation could rapidly inhibit AMPK activation through increased AMPK phosphorylation at the inhibitory Ser485/491 residues without a change at the activating Th172 residue, and this was accompanied by a reduction of ACC phosphorylation. Using specific inhibitors and gene silencing, we found that such AMPK inhibition involved a signaling cascade through Lyn-Syk-Akt. When AMPK was activated by AICAR, A769662 and metformin, FcεRI-mediated Syk, ERK, JNK and p38 activation, and TNFα release were all inhibited. Consistently, AMPK inhibition by compound C increased FcεRI-mediated Lyn activation. Moreover, AMPK activation dominantly impaired IgE-induced recruitment of signal proteins to the FcεRI by blocking the formation of FcεRIβ-Lyn-Syk, FcεRIγ-Lyn-Syk, and AMPK-FcεRIβ complexes. In vitro kinase assay further revealed the ability of AMPKα2 to phosphorylate FcεRIβ in the complex. In vivo, AMPK activation by metformin could readily reduce vascular permeability and ear swelling in a mouse model of passive cutaneous anaphylaxis mediated by IgE. In summary, our findings demonstrate that IgE-mediated FcεRI activation results in AMPK inhibition through activation of Lyn-Syk-Akt pathway, and as such FcεRI receptor can efficiently propagate Lyn-mediated allergic signaling and response. These results provide important insights into the use of AMPK activators for the treatment of allergic diseases.

Key messages

  • AMPK is inhibited by FcεRI via Lyn-Syk-Akt signaling in RBL-2H3 cells.

  • AMPK inhibition supports FcεRI-mediated Lyn signaling and allergic response.

  • Metformin has inhibitory effect on passive cutaneous anaphylaxis.


AMPK FcεRI Syk Akt Basophils Anaphylaxis 







AMP-activated protein kinase


Bone marrow-derived mast cells


2,4-Dinitrophenylated bovine serum albumin


Fcepsilon receptor I


Glutathione S-transferase


Human serum albumin


Immunoglobulin E


Immunoreceptor tyrosine-based activation motif


Linker for activation of T cell


Liver kinase B1


Mitogen-activated protein kinase


Passive cutaneous anaphylaxis


Phospholipase Cγ


Protein tyrosine kinase


Src family kinase


Small interfering RNA


Spleen tyrosine kinase



This work was supported by a grant from the Ministry of Science and Technology (MOST 103-2320-B-002-069 -MY3).

Conflict of interest

The authors declare that they have no competing interests.

Supplementary material

109_2015_1339_MOESM1_ESM.pdf (264 kb)
ESM 1 (PDF 264 kb)


  1. 1.
    Stone KD, Prussin C, Metcalfe DD (2010) IgE, mast cells, basophils, and eosinophils. J Allergy Clin Immunol 125:S73–S80PubMedCentralCrossRefPubMedGoogle Scholar
  2. 2.
    Wu LC (2011) Immunoglobulin E receptor signaling and asthma. J Biol Chem 286:32891–32897PubMedCentralCrossRefPubMedGoogle Scholar
  3. 3.
    Wedemeyer J, Galli SJ (2000) Mast cells and basophils in acquired immunity. Br Med Bull 56:936–955CrossRefPubMedGoogle Scholar
  4. 4.
    Akdis M, Akdis CA (2009) Therapeutic manipulation of immune tolerance in allergic disease. Nat Rev Drug Discov 8:645–660CrossRefPubMedGoogle Scholar
  5. 5.
    Kinet JP (1999) The high-affinity IgE receptor (Fc epsilon RI): from physiology to pathology. Annu Rev Immunol 17:931–972CrossRefPubMedGoogle Scholar
  6. 6.
    Turner H, Kinet JP (1999) Signalling through the high-affinity IgE receptor Fc epsilonRI. Nature 402:B24–B30CrossRefPubMedGoogle Scholar
  7. 7.
    Wilson BS, Pfeiffer JR, Oliver JM (2000) Observing FcepsilonRI signaling from the inside of the mast cell membrane. J Cell Biol 149:1131–1142PubMedCentralCrossRefPubMedGoogle Scholar
  8. 8.
    Kawakami T, Galli SJ (2002) Regulation of mast-cell and basophil function and survival by IgE. Nat Rev Immunol 2:773–786CrossRefPubMedGoogle Scholar
  9. 9.
    Sada K, Takano T, Yanagi S, Yamamura H (2001) Structure and function of Syk protein-tyrosine kinase. J Biochem 130:177–186CrossRefPubMedGoogle Scholar
  10. 10.
    Ewart MA, Kennedy S (2011) AMPK and vasculoprotection. Pharmacol Ther 131:242–253CrossRefPubMedGoogle Scholar
  11. 11.
    Viollet B, Horman S, Leclerc J, Lantier L, Foretz M, Billaud M, Giri S, Andreelli F (2010) AMPK inhibition in health and disease. Crit Rev Biochem Mol Biol 45:276–295PubMedCentralCrossRefPubMedGoogle Scholar
  12. 12.
    Kovacic S, Soltys CL, Barr AJ, Shiojima I, Walsh K, Dyck JR (2003) Akt activity negatively regulates phosphorylation of AMP-activated protein kinase in the heart. J Biol Chem 278:39422–39427CrossRefPubMedGoogle Scholar
  13. 13.
    Horman S, Vertommen D, Heath R, Neumann D, Mouton V, Woods A, Schlattner U, Wallimann T, Carling D, Hue L et al (2006) Insulin antagonizes ischemia-induced Thr172 phosphorylation of AMP-activated protein kinase alpha-subunits in heart via hierarchical phosphorylation of Ser485/491. J Biol Chem 281:5335–5340CrossRefPubMedGoogle Scholar
  14. 14.
    Berggreen C, Gormand A, Omar B, Degerman E, Goransson O (2009) Protein kinase B activity is required for the effects of insulin on lipid metabolism in adipocytes. Am J Physiol Endocrinol Metab 296:E635–E646CrossRefPubMedGoogle Scholar
  15. 15.
    Ning J, Xi G, Clemmons DR (2011) Suppression of AMPK activation via S485 phosphorylation by IGF-I during hyperglycemia is mediated by AKT activation in vascular smooth muscle cells. Endocrinology 152:3143–3154PubMedCentralCrossRefPubMedGoogle Scholar
  16. 16.
    Hardie DG (2014) AMP-activated protein kinase: a key regulator of energy balance with many roles in human disease. J Intern Med 276:543–559, See comment in PubMed Commons below CrossRefGoogle Scholar
  17. 17.
    Hwang SL, Li X, Lu Y, Jin Y, Jeong YT, Kim YD, Lee IK, Taketomi Y, Sato H, Cho YS et al (2013) AMP-activated protein kinase negatively regulates FcepsilonRI-mediated mast cell signaling and anaphylaxis in mice. J Allergy Clin Immunol 132:729–736, e12 CrossRefPubMedGoogle Scholar
  18. 18.
    Hwang SL, Lu Y, Li X, Kim YD, Cho YS, Jahng Y, Son JK, Lee YJ, Kang W, Taketomi Y et al (2014) ERK1/2 antagonize AMPK-dependent regulation of FceRI mediated mast cell activation and anaphylaxis. J Allergy Clin Immunol 134:714–721CrossRefPubMedGoogle Scholar
  19. 19.
    Blake RA, Broome MA, Liu X, Wu J, Gishizky M, Sun L, Courtneidge SA (2000) SU6656, a selective src family kinase inhibitor, used to probe growth factor signaling. Mol Cell Biol 20:9018–9027PubMedCentralCrossRefPubMedGoogle Scholar
  20. 20.
    Lee JH, Mun SH, Ko NY, Kim JW, Kim DK, Kim JD, Her E, Choi WS (2007) Roles of Src-family kinase isoforms, Lyn, Fyn, Fgr, and c-Src on degranulation in RBL-2H3 mast cells. J Life Sci 17:350–355CrossRefGoogle Scholar
  21. 21.
    Sanderson MP, Wex E, Kono T, Uto K, Schnapp A (2010) Syk and Lyn mediate distinct Syk phosphorylation events in FcɛRI-signal transduction: Implications for regulation of IgE-mediated degranulation. Mol Immunol 48:171–178CrossRefPubMedGoogle Scholar
  22. 22.
    Kovarova M, Tolar P, Arudchandran R, Draberova L, Rivera J, Draber P (2001) Structure-function analysis of Lyn kinase association with lipid rafts and initiation of early signaling events after Fcepsilon receptor I aggregation. Mol Cell Biol 21:8318–8328PubMedCentralCrossRefPubMedGoogle Scholar
  23. 23.
    Vonakis BM, Haleem-Smith H, Benjamin P, Metzger H (2001) Interaction between the unphosphorylated receptor with high affinity for IgE and Lyn kinase. J Biol Chem 276:1041–1050CrossRefPubMedGoogle Scholar
  24. 24.
    Kimura T, Kihara H, Bhattacharyya S, Sakamoto H, Appella E, Siraganian RP (1996) Downstream signaling molecules bind to different phosphorylated immunoreceptor tyrosine-based activation motif (ITAM) peptides of the high affinity IgE receptor. J Biol Chem 271:27962–27968CrossRefPubMedGoogle Scholar
  25. 25.
    Simons K, Toomre D (2000) Lipid rafts and signal transduction. Nat Rev Mol Cell Biol 1:31–39CrossRefPubMedGoogle Scholar
  26. 26.
    Mocsai A, Ruland J, Tybulewicz VL (2010) The SYK tyrosine kinase: a crucial player in diverse biological functions. Nat Rev Immunol 10:387–402CrossRefPubMedGoogle Scholar
  27. 27.
    Liu WM, Huang P, Kar N, Burgett M, Muller-Greven G, Nowacki AS, Distelhorst CW, Lathia JD, Rich JN, Kappes JC, Gladson CL (2013) Lyn facilitates glioblastoma cell survival under conditions of nutrient deprivation by promoting autophagy. PLoS One 8:e70804Google Scholar
  28. 28.
    Calixto MC, Lintomen L, André DM, Leiria LO, Ferreira D, Lellis-Santos C, Anhê GF, Bordin S, Landgraf RG, Antunes E (2013) Metformin attenuates the exacerbation of the allergic eosinophilic inflammation in high fat-diet-induced obesity in mice. PLoS One 8:e76786Google Scholar
  29. 29.
    Stapleton D, Mitchelhill KI, Gao G, Widmer J, Michell BJ, Teh T, House CM, Fernandez CS, Cox T, Witters LA et al (1996) Mammalian AMP-activated protein kinase subfamily. J Biol Chem 271:611–614CrossRefPubMedGoogle Scholar
  30. 30.
    Quentin T, Kitz J, Steinmetz M, Poppe A, Bar K, Kratzner R (2011) Different expression of the catalytic alpha subunits of the AMP activated protein kinase—an immunohistochemical study in human tissue. Histol Histopathol 26:589–596PubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Kai-Chun Lin
    • 1
  • Duen-Yi Huang
    • 1
  • De-Wei Huang
    • 1
  • Shiang-Jong Tzeng
    • 1
  • Wan-Wan Lin
    • 1
    • 2
  1. 1.Department of Pharmacology, College of MedicineNational Taiwan UniversityTaipeiTaiwan
  2. 2.Graduate Institute of Medical SciencesTaipei Medical UniversityTaipeiTaiwan

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