Loxoprofen enhances intestinal barrier function via generation of its active metabolite by carbonyl reductase 1 in differentiated Caco-2 cells

Satoshi Endo; Tsubasa Nishiyama; Tomoe Matuoka; Takeshi Miura; Toru Nishinaka; Toshiyuki Matsunaga; Akira Ikari

doi:10.1016/j.cbi.2021.109634

Loxoprofen enhances intestinal barrier function via generation of its active metabolite by carbonyl reductase 1 in differentiated Caco-2 cells

Chem Biol Interact. 2021 Oct 1:348:109634. doi: 10.1016/j.cbi.2021.109634. Epub 2021 Sep 8.

Authors

Satoshi Endo¹, Tsubasa Nishiyama¹, Tomoe Matuoka¹, Takeshi Miura², Toru Nishinaka³, Toshiyuki Matsunaga⁴, Akira Ikari⁵

Affiliations

¹ Laboratory of Biochemistry, Department of Biopharmaceutical Sciences, Gifu Pharmaceutical University, 501-1196, Gifu, Japan.
² Pharmaceutical Education Support Center, School of Pharmacy and Pharmaceutical Sciences, Mukogawa Women's University, Nishinomiya, 663-8184, Japan.
³ Laboratory of Biochemistry, Faculty of Pharmacy, Osaka Ohtani University, Tondabayashi, 584-8540, Japan.
⁴ Education Center of Green Pharmaceutical Sciences, Gifu Pharmaceutical University, Gifu, 502-8585, Japan.
⁵ Laboratory of Biochemistry, Department of Biopharmaceutical Sciences, Gifu Pharmaceutical University, 501-1196, Gifu, Japan. Electronic address: ikari@gifu-pu.ac.jp.

PMID: 34506768
DOI: 10.1016/j.cbi.2021.109634

Abstract

Nonsteroidal anti-inflammatory drugs (NSAIDs) are used worldwide as antipyretic analgesics and agents for rheumatoid arthritis and osteoarthritis, but known to cause damage to the gastrointestinal mucosae as their serious adverse effects. Few studies showed the impairment of intestinal epithelial barrier function (EBF) by high concentrations (0.5-1 mM) of NSAIDs, but the underlying mechanism is not fully understood. This study is aimed at clarifying effects at a low concentration (50 μM) of three NSAIDs, loxoprofen (Lox), ibuprofen and indomethacin, on intestinal EBF using human intestinal epithelial-like Caco-2 cells. Among those NSAIDs, Lox increased the transepithelial electric resistance (TER) value, decreased the paracellular Lucifer yellow CH (LYCH) permeability, and upregulated claudin (CLDN)-1, -3 and -5, indicating that low doses of Lox enhanced EBF through increasing expression of CLDNs. Lox is known to be metabolized to a pharmacologically active metabolite, (2S,1'R,2'S)-loxoprofen alcohol (Lox-RS), by carbonyl reductase 1 (CBR1), which is highly expressed in human intestine. CBR1 was expressed in the Caco-2 cells, and the pretreatment with a CBR1 inhibitor suppressed both the Lox-evoked CLDN upregulation and EBF enhancement. In addition, the treatment of the cells with Lox-RS resulted in higher TER value and lower LYCH permeability than those with Lox. Thus, Lox-RS synthesized by CBR1 may greatly contribute to the improving efficacy of Lox on the barrier function. Since EBF is decreased in inflammatory bowel disease, we finally examined the effect of Lox on EBF using the Caco-2/THP-1 co-culture system, which is used as an in vitro inflammatory bowel disease model. Lox significantly recovered EBF which was impaired by inflammatory cytokines secreted from THP-1 macrophages. These in vitro observations suggest that Lox enhances intestinal EBF, for which the metabolism of Lox to Lox-RS by CBR1 has an important role.

Keywords: Carbonyl reductase 1; Claudin; Epithelial barrier function; Loxoprofen; Nonsteroidal anti-inflammatory drug.

MeSH terms

Anti-Inflammatory Agents, Non-Steroidal / metabolism
Anti-Inflammatory Agents, Non-Steroidal / pharmacology*
Caco-2 Cells
Carbonyl Reductase (NADPH) / metabolism*
Cell Differentiation / drug effects*
Cytokines / metabolism
Humans
Intestinal Mucosa / cytology
Intestinal Mucosa / drug effects*
Intestinal Mucosa / metabolism*
Phenylpropionates / metabolism
Phenylpropionates / pharmacology*

Substances

Anti-Inflammatory Agents, Non-Steroidal
Cytokines
Phenylpropionates
loxoprofen
Carbonyl Reductase (NADPH)