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22 May 2018 : Laboratory Research  

Peptidoglycan Suppresses Phagocytic Activities and Apoptosis of Macrophages in Colonic Mucosa Tissues of Crohn’s Disease Patients and In Vitro

Yang Jing1ABCDEF*, Ying Ran1ABCDF, Jing Zhao1BCF, Zhe Zhou1CF, Jun Zhang1B, Yiqi Qian1B, Zhiqi Yin2B, Mingfang Zhang2B, Zongshun Lv1F, Lu Zhou1AFG, Bangmao Wang1ACEFG

DOI: 10.12659/MSM.910266

Med Sci Monit 2018; 24: LBR3382-3392

Abstract

BACKGROUND: Rac1 signaling plays a crucial role in controlling macrophage functions in CD. Peptidoglycan triggers several intracellular signaling pathways, including activation of Rac1, to regulate the function of macrophage. Suppressed Rac1 signaling in non-inflamed colonic mucosa of Crohn’s disease patients has been shown to correlate with increased innate immunity.

MATERIAL AND METHODS: We examined the effect of peptidoglycan on Rac1 signaling in macrophages and mucosal tissue samples collected from 10 patients with active Crohn’s disease and further investigated the effects of peptidoglycan on apoptosis and phagocytic activities of macrophages in vitro.

RESULTS: Macrophage infiltration and Rac1 signaling was increased in inflamed mucosal tissues of Crohn’s disease patients. Immunoblotting assays revealed that peptidoglycan dose- and time-dependently increased the expression of Rac1-GTP, phosphorylated VAV1, and phosphorylated PAK1in RAW264.7 macrophages, which, however, was attenuated by 6-thioguanine. Peptidoglycan also dose-dependently inhibited phagocytic activities of human peripheral blood monocytic cells (PBMCs), which were partially abated by 6-thioguanine or NSC23766. Flow cytometry showed that peptidoglycan (3 μg/mL) decreased the proportion of apoptotic human PBMCs versus controls. The addition of 6-thioguanine or NSC3766 to peptidoglycan led to a sharper rise in the proportion of apoptotic human PBMCs than 6-thioguanine or NSC3766 alone.

CONCLUSIONS: Our findings suggest that Rac1 signaling is a common molecular target shared by peptidoglycan and immunosuppressive treatment in intestinal macrophages. Inhibiting Rac1 activation may be crucial for optimizing macrophage immunity for treatment of Crohn’s disease.

Keywords: Crohn Disease, Immunosuppression, Macrophages, Peptidoglycan, rac1 GTP-Binding Protein

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750