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Cis 9, trans 11, but not trans 10, cis 12 CLA isomer, impairs intestinal epithelial barrier function in IPEC-J2 cells and mice through activation of GPR120-[Ca2+]i and MLCK signaling pathway

Abstract

This study aimed to investigate the effects of conjugated linoleic acid (CLA) on intestinal epithelial barrier function and explore the underlying mechanisms. IPEC-J2 cells and mice were treated with different CLA isomers. The intestinal epithelial barrier function determined by transepithelial electrical resistance (TEER), the expression of tight junction proteins, and the involvement of G-protein coupled receptor 120 (GPR120), intracellular calcium ([Ca2+]i) and myosin light chain kinase (MLCK) were assessed. In vitro, c9, t11-CLA, but not t10, c12-CLA isomer, impaired epithelial barrier function in IPEC-J2 by downregulating expression of tight junction proteins. Meanwhile, c9, t11-CLA isomer enhanced GPR120 expression, while knockdown of GPR120 eliminated the impaired epithelial barrier function induced by c9, t11-CLA isomer. In addition, c9, t11-CLA isomer increased [Ca2+]i and activated MLCK signaling pathway in a GPR120-dependent manner. However, chelation of [Ca2+]i reversed c9, t11-CLA isomer-induced MLCK activation and epithelial barrier function impairment of IPEC-J2. Furthermore, inhibition of MLCK totally abolished the impairment of epithelial barrier function induced by c9, t11-CLA. In vivo, dietary supplementation with c9, t11-CLA rather than t10, c12-CLA isomer decreased expression of intestinal tight junction proteins and GPR120, increased intestinal permeability, and activated MLCK signaling pathway in mice. Taken together, our findings showed that c9, t11-CLA, but not t10, c12-CLA isomer, impaired intestinal epithelial barrier function in IPEC-J2 cells and mice through activation of GPR120-[Ca2+]i and MLCK signaling pathway. These data provided new insight into the regulation of intestinal epithelial barrier by different CLA isomers and more reference for CLA application in human and animals.

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Article information


Accepted
21 Mar 2020
First published
23 Mar 2020

Food Funct., 2020, Accepted Manuscript
Article type
Paper

Cis 9, trans 11, but not trans 10, cis 12 CLA isomer, impairs intestinal epithelial barrier function in IPEC-J2 cells and mice through activation of GPR120-[Ca2+]i and MLCK signaling pathway

H. Su, W. Zhao, F. Zhang, M. Song, F. Liu, J. Zheng, M. Ling, X. Yang, Q. Yang, H. He, L. Chen, X. Lai, X. Zhu, L. Wang, P. Gao, G. Shu, Q. Jiang and S. Wang, Food Funct., 2020, Accepted Manuscript , DOI: 10.1039/D0FO00376J

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