6-Mercaptopurine reduces macrophage activation and gut epithelium proliferation through inhibition of GTPase Rac1

• Published in: Inflammatory bowel diseases Vol. 20; no. 9; pp. 1487 - 1495
• Main Authors: Marinković, Goran, Hamers, Anouk A J, de Vries, Carlie J M, de Waard, Vivian
• Format: Journal Article
• Language: English
• Published: England 01.09.2014
• Subjects: Animals; Cell Proliferation - drug effects; Cells, Cultured; Chemokine CCL2 - genetics; Chemokine CCL2 - metabolism; Epithelial Cells - cytology; Epithelial Cells - drug effects; Epithelial Cells - metabolism; Gastrointestinal Tract - cytology; Gastrointestinal Tract - drug effects; Gastrointestinal Tract - metabolism; Humans; Immunosuppressive Agents - pharmacology; Interleukin-8 - genetics; Interleukin-8 - metabolism; Macrophage Activation - drug effects; Macrophages - cytology; Macrophages - drug effects; Macrophages - metabolism; MAP Kinase Kinase 4 - genetics; MAP Kinase Kinase 4 - metabolism; Mercaptopurine - pharmacology; Mice; Nitric Oxide Synthase Type II - genetics; Nitric Oxide Synthase Type II - metabolism; Phosphorylation; rac1 GTP-Binding Protein - antagonists & inhibitors; rac1 GTP-Binding Protein - genetics; rac1 GTP-Binding Protein - metabolism; Signal Transduction; STAT3 Transcription Factor - genetics; STAT3 Transcription Factor - metabolism; Tumor Necrosis Factor-alpha
• ISSN: 1078-0998; 1536-4844
• DOI: 10.1097/mib.0000000000000122

Inflammatory bowel disease is characterized by chronic intestinal inflammation. Azathioprine and its metabolite 6-mercaptopurine (6-MP) are effective immunosuppressive drugs that are widely used in patients with inflammatory bowel disease. However, established understanding of their immunosuppressive mechanism is limited. Azathioprine and 6-MP have been shown to affect small GTPase Rac1 in T cells and endothelial cells, whereas the effect on macrophages and gut epithelial cells is unknown. Macrophages (RAW cells) and gut epithelial cells (Caco-2 cells) were activated by cytokines and the effect on Rac1 signaling was assessed in the presence or absence of 6-MP. Rac1 is activated in macrophages and epithelial cells, and treatment with 6-MP resulted in Rac1 inhibition. In macrophages, interferon-γ induced downstream signaling through c-Jun-N-terminal Kinase (JNK) resulting in inducible nitric oxide synthase (iNOS) expression. iNOS expression was reduced by 6-MP in a Rac1-dependent manner. In epithelial cells, 6-MP efficiently inhibited tumor necrosis factor-α-induced expression of the chemokines CCL2 and interleukin-8, although only interleukin-8 expression was inhibited in a Rac1-dependent manner. In addition, activation of the transcription factor STAT3 was suppressed in a Rac1-dependent fashion by 6-MP, resulting in reduced proliferation of the epithelial cells due to diminished cyclin D1 expression. These data demonstrate that 6-MP affects macrophages and gut epithelial cells beneficially, in addition to T cells and endothelial cells. Furthermore, mechanistic insight is provided to support development of Rac1-specific inhibitors for clinical use in inflammatory bowel disease.