l-Glutamine Attenuates DSS-Induced Colitis via Induction of MAPK Phosphatase-1

Soo-Yeon Jeong, Yoo Na Im, Ji Young Youm, Hern-Ku Lee, Suhn-Young Im, Soo-Yeon Jeong, Yoo Na Im, Ji Young Youm, Hern-Ku Lee, Suhn-Young Im

Abstract

Inflammatory bowel disease (IBD), encompassing ulcerative colitis and Crohn's disease, is a multifactorial inflammatory disease of the small intestine and colon. Many investigators have reported that l-glutamine (Gln) therapy improves outcomes of experimental colitis models, although the mechanism is not fully understood. Regarding the anti-inflammatory properties of Gln, we have shown that Gln can effectively deactivate cytosolic phospholipase A₂ (cPLA₂) by rapid induction of MAPK phosphatase (MKP)-1. In this study, we explore the possibility that Gln ameliorates dextran sulfate sodium (DSS)-induced colitis via MKP-1 induction, resulting in inhibition of cPLA₂, which has been reported to play a key role in the pathogenesis of IBD. Oral Gln intake attenuated DSS-induced colitis. Gln inhibited cPLA₂ phosphorylation, as well as colonic levels of TNF-α and leukotriene (LT)B₄. Gln administration resulted in early and enhanced MKP-1 induction. Importantly, MKP-1 small interfering RNA (siRNA), but not control siRNA, significantly abrogated the Gln-mediated (1) induction of MKP-1; (2) attenuation of colitis (colon length, histological abnormality, and inflammation; and (3) inhibition of cPLA₂ phosphorylation and colonic levels of TNF-α and LTB₄. These data indicated that Gln ameliorated DSS-induced colitis via MKP-1 induction.

Keywords: DSS; MKP-1; cPLA2; glutamine; inflammatory bowel disease.

Conflict of interest statement

The authors declare that there is no conflict of interests.

Figures

Figure 1
Figure 1
Oral l-glutamine (Gln) intake improves DSS-induced acute colitis. (A) Disease severity; (B) the colon length on day 5; (C) histological examination on day 5; (D) histological score; (E,F) colonic levels of TNF-α and LTB4 at 24 h; Data in (A,E,F) represent the means ± SEM of three independent experiments (n = 5 mice/group/time point); (B–D) A representative from three independent experiments (n = 3 mice/group/time point) is shown. Scale bar = 50 μm. * p < 0.001 vs. vehicle group; ** p < 0.05 vs. DSS group in (A,D–F).
Figure 2
Figure 2
Gln enhances MKP-1 induction in colitis. (A,B) Immunoblot analysis of MKP-1. Data are representative of three independent experiments (n = 3 mice/group/time point). * p < 0.05, ** p < 0.01 vs. DSS only group in (A); * p < 0.05 vs. DSS + Gln group in (B).
Figure 3
Figure 3
Gln ameliorates colitis via MKP-1 induction. (A) Disease severity; (B) the colon length on day 5; (C) histological examination on day 5; (D) histological score; (E,F) colonic levels of TNF-α and LTB4; Data in (A,E,F) represent the means ± SEM of three independent experiments (n = 5 mice/group/time point); (B,C) A representative from three independent experiments (n = 3 mice/group/time point) is shown. Scale bar = 50 μm. * p < 0.001 vs. vehicle group; ** p < 0.05 vs. DSS group; #p < 0.05 vs. DSS + Gln group in (A,D–F).
Figure 4
Figure 4
cPLA2 inhibition is associated with the beneficial effect of Gln. (A,B) Immunoblot analysis of cPLA2 phosphorylation; (C) disease severity; (D) histological abnormality; (E) histological score; (F) colon length; colonic levels of TNF-α (G) and LTB4 (H); (A,B) A representative of three independent experiments (n = 3 mice/group/time point). * p < 0.05 vs. DSS only group in (A); * p < 0.05 vs. DSS + Gln group in (B); Data in (C,G,H) represent the means ± SEM of three independent experiments (n = 5 mice/group/time point); (D–F) A representative from three independent experiments (n = 3 mice/group/time point) is shown. Scale bar = 50 μm. * p < 0.001 vs. vehicle group; ** p < 0.05 vs. DSS group in (C,E,G,H).
Figure 5
Figure 5
Anti-inflammatory action mechanisms of Gln. Gln binds to G-protein coupled receptors (GPCRs), an allosteric receptor, (unpublished data), and increases ERK activity via activation of the pathway involving Ca2+/Ras/c-Raf/MEK (ERK cascade) [41]. ERK phosphorylates MKP-1 on two carboxyl-terminal serine residues—serine 359 and serine 364, which enhances MKP-1 stabilization, resulting in the early induction of MKP-1 [20]. MKP-1 deactivates cPLA2 either by dephosphorylating p38 [20,23], which is a major upstream pathway for cPLA2 phosphorylation, or by directly dephosphorylating cPLA2 due to enhanced physical interaction between Gln-induced MKP-1 and cPLA2 [24]. Deactivation of p38 and cPLA2 results in suppression of many cardinal inflammatory mediators including reactive oxygen species.

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