Cyclosporine protects from intestinal epithelial injury by modulating butyrate uptake via upregulation of membrane monocarboxylate transporter 1 levels

Shinji Ota; Hirotake Sakuraba; Hiroto Hiraga; Shukuko Yoshida; Miwa Satake; Yui Akemoto; Nahoko Tanaka; Rina Watanabe; Maeda Takato; Yasuhisa Murai; Kayo Ueno; Takenori Niioka; Makoto Hayakari; Yoh Ishiguro; Shinsaku Fukuda

doi:10.1016/j.bbrep.2020.100811

Cyclosporine protects from intestinal epithelial injury by modulating butyrate uptake via upregulation of membrane monocarboxylate transporter 1 levels

Biochem Biophys Rep. 2020 Oct 21:24:100811. doi: 10.1016/j.bbrep.2020.100811. eCollection 2020 Dec.

Authors

Affiliations

¹ Department of Gastroenterology and Hematology, Hirosaki University Graduate School of Medicine, Hirosaki, Japan.
² Shibata Irika Co.Ltd.Hirosaki, Japan.
³ Division of Pharmaceutical Science, Hirosaki University Hospital, Hirosaki, Japan.
⁴ Division of Clinical Research, Hirosaki National Hospital, National Hospital Organization, Hirosaki, Japan.

Abstract

Background and aims: A relationship between treatment outcomes and intestinal microbiota in patients with inflammatory bowel diseases has been demonstrated. Cyclosporine treatment leads to rapid improvement in severe ulcerative colitis. We hypothesized that the potent effects of cyclosporine would be exerted through relationships between intestinal epithelial cells (IECs) and the host microbiota. The present study was designed to elucidate the effects of cyclosporine on monocarboxylate transporter 1 (MCT1) regulation and butyrate uptake by IECs.

Methods: Colitis was induced in C57BL6 mice via the administration of 4% dextran sulfate sodium in drinking water, following which body weights, colon lengths, and histological scores were evaluated. To examine the role of butyrate in the protective effects of cyclosporine, MCT1 inhibitor and an antibiotic cocktail was administered and tributyrin (TB; a prodrug of butyrate) was supplemented; MCT1 protein expression and acetylated histone 3 (AcH3) signals in IECs, as well as the MCT1-membrane fraction of Caco-2 cells, were evaluated. To explore butyrate uptake, as s butyrate derivatives, 3-bromopyruvic acid (3-BrPA) and 1-pyrenebutyric acid were used.

Results: Treatment with cyclosporine inhibited body weight loss and colon length shortening. However, treatment with MCT1 inhibitor and the antibiotic cocktail negated the efficacy of cyclosporine, whereas TB supplementation restored its protective effect. Furthermore, cyclosporine upregulated MCT1 expression in the membrane and the AcH3 signal in IECs, while also inducing higher anti-inflammatory cytokine production compared to that in the vehicle-treated mice. The transcription level of MCT1 mRNA in IECs and Caco-2 cells did not increase with cyclosporine treatment; however, cyclosporine treatment increased membrane MCT1 expression in these cells and uptake of butyrate derivative.

Conclusion: Cyclosporine treatment modulates butyrate uptake via the post-transcriptional upregulation of membrane MCT1 levels in IECs.

Keywords: Butyrate; Colitis; Cyclosporine; Intestinal epithelial cells; Monocarboxylate transporter 1; acetylated histone 3, AcH3; antibiotic cocktail, abx; dextran sulfate sodium, DSS; inflammatory bowel disease, IBD; intestinal epithelial cells, IECs; regulatory T cells, Tregs; short-chain fatty acids, SCFAs; tributyrin, TB; ulcerative colitis, UC.