Glutamine up-regulates MAPK phosphatase-1 induction via activation of Ca2+→ ERK cascade pathway

Otgonzaya Ayush; Zhe Wu Jin; Hae-Kyoung Kim; Yu-Rim Shin; Suhn-Young Im; Hern-Ku Lee

doi:10.1016/j.bbrep.2016.05.011

Glutamine up-regulates MAPK phosphatase-1 induction via activation of Ca²⁺→ ERK cascade pathway

Biochem Biophys Rep. 2016 May 12:7:10-19. doi: 10.1016/j.bbrep.2016.05.011. eCollection 2016 Sep.

Authors

Otgonzaya Ayush¹, Zhe Wu Jin², Hae-Kyoung Kim³, Yu-Rim Shin⁴, Suhn-Young Im⁵, Hern-Ku Lee³

Affiliations

¹ Department of Dermatology, Medical University, Mongolian National University of Medical Sciences, Ulaanbaatar, Mongolia.
² Department of Anatomy and Histology and Embryology, Yanbian University Medical College, YanJi City, Jilin Province, China.
³ Departments of Immunology and Institute for Medical Science, Chonbuk National University Medical School, Jeonju, Republic of Korea.
⁴ Biofoods Story, Inc, 477 Jeonjucheon-seoro, Wansan-gu, Jeonju, Jeonbuk 560-821, Republic of Korea.
⁵ Department of Biological Sciences, College of Natural Sciences, Chonnam National University, Gwangju, Republic of Korea.

Abstract

The non-essential amino acid L-glutamine (Gln) displays potent anti-inflammatory activity by deactivating p38 mitogen activating protein kinase and cytosolic phospholipase A₂ via induction of MAPK phosphatase-1 (MKP-1) in an extracellular signal-regulated kinase (ERK)-dependent way. In this study, the mechanism of Gln-mediated ERK-dependency in MKP-1 induction was investigated. Gln increased ERK phosphorylation and activity, and phosphorylations of Ras, c-Raf, and MEK, located in the upstream pathway of ERK, in response to lipopolysaccharidein vitro and in vivo. Gln-induced dose-dependent transient increases in intracellular calcium ([Ca²⁺]_i) in MHS macrophage cells. Ionomycin increased [Ca²⁺]_i and activation of Ras → ERK pathway, and MKP-1 induction, in the presence, but not in the absence, of LPS. The Gln-induced pathways involving Ca²⁺→ MKP-1 induction were abrogated by a calcium blocker. Besides Gln, other amino acids including L-phenylalanine and l-cysteine (Cys) also induced Ca²⁺ response, activation of Ras → ERK, and MKP-1 induction, albeit to a lesser degree. Gln and Cys were comparable in suppression against 2, 4-dinitrofluorobenzene-induced contact dermatitis. Gln-mediated, but not Cys-mediated, suppression was abolished by MKP-1 small interfering RNA. These data indicate that Gln induces MKP-1 by activating Ca²⁺→ ERK pathway, which plays a key role in suppression of inflammatory reactions.

Keywords: AP-1, activating protein 1; Ala, alanine; Asp, aspartate; BAPTA, 1,2-bis(o-aminophenoxy)ethane-N,N,N’,N’-tetraacetic acid tetraacetoxymethylester; CD, contact dermatitis; CaM, calmodulin; CaR, Ca2+-sensing receptor; DMSO, dimethyl sulfoxide; DNFB, 1-fluoro-2,4-dinitrobenzene; ERK, extracellular signal-regulated kinase; ESR, ear swelling response; Gln, L-glutamine; Glu, glutamate; Gly, glycine; H&E, hematoxylin and eosin; JNK, c-Jun N-terminal kinase; L-Glutamine; LPS, lipopolysaccharides; MAPK Phosphatase-1; MAPK, mitogen activated protein kinase; MKP-1, MAPK phosphatase-1; Mitogen-activated protein kinase; PEI, polyethyleneimine; Ras/c-Raf/MEK/ERK, extracellular-signal-regulated kinase; [Ca2+]i, intracellular calcium concentration; cPLA2, cytoplasmic phospholipase A2; siRNA, small interfering RNA.