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Effects of NOD2 in intestinal epithelial cells: Cross‐talk between NHE and Na + ‐K + ‐ATPase activity
Author(s) -
Cabral Jose,
Grácio Daniela,
SoaresdaSilva Patricio,
Magro Fernando
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.969.7
Subject(s) - chemistry , protein kinase a , adenylyl cyclase , tyrosine kinase , kinase , atpase , signal transduction , microbiology and biotechnology , biochemistry , biology , enzyme
NOD‐like receptors are a family of pattern recognition receptors that plays a role in the control of inammation and have been associated with inflammatory bowel disease (IBD). Na + /H + exchangers (NHE) are responsible for the net uptake of sodium, electrolyte homeostasis and decreases in pH i during inflammatory process. Aim Evaluate the effect of NOD2 upon NHE and Na + ‐K + ‐ATPase activity. Methods NHE activity, Na + ‐K + ‐ATPase activity and levels of intracellular calcium were measured using fluorimetric assays. Results Short‐term NOD2 activation stimulated NHE1 activity in a concentration dependent manner (100±3 to 129±4% in T84 and 100±4 to 133±2% in NCM460 cells). To understand the transduction signalling involved experiments were performed in the presence of inhibitors of Rip‐like interacting caspase‐like apoptosis‐regulatory protein kinase (RIP2), NF‐kβ, tyrosine‐protein kinase (SRC), transforming growth factor‐beta‐activated kinase1 (TAK 1), TNF receptor associated fator 6, adenylyl cyclase (AC), protein kinase A (PKA), phospholipase C (PLC), protein kinase C, Na + ‐K + ‐ATPase and potassium channels blockers. The NOD2‐mediated increase in NHE1 activity was prevented by inhibitors of RIP2, SRC, TAK1, AC, PKA, PLC, Na + ‐K + ‐ATPase and Ca 2+ ‐dependent‐ATP‐sensitive K + channels in both cell lines; this pathway was NF‐kβ independent. Increases in intracellular Ca 2+ were observed after NOD2 activation (100±1 to 360±1% in T84 and 100±1 to 254±12% in NCM460 cells). Conclusions Activation of NOD2 promotes an increase in Na + ‐K + ‐ATPase and NHE1 activities as a result of opening of Ca 2+ ‐dependent ATP‐sensitive K + channels. A better understanding of this pathway is considered for the development of novel therapies for IBD.