Kinetics of manganese transport and gene expressions of manganese transport carriers in Caco-2 cell monolayers
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Kinetics of manganese transport and gene expressions of manganese transport carriers in Caco-2 cell monolayers Xiaoli Li • Jingjing Xie • Lin Lu • Liyang Zhang Lingyan Zhang • Yaxue Zou • Qiuyue Wang • Xugang Luo • Sufen Li
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Received: 20 May 2013 / Accepted: 21 August 2013 Ó Springer Science+Business Media New York 2013
Abstract Two experiments were conducted to investigate the kinetics of manganese (Mn) transport in Caco-2 cell monolayers and the gene expressions of Mn transport carriers in apical (AP) and basolateral (BL) membranes. In experiment 1, the cells were treated with the medium containing 146 lmol/L of Mn (MnSO4H2O). Both the uptake and transport of Mn from AP–BL or from BL–AP at different timepoints were assessed to determine the optimal time for kinetics of Mn transport. The transport of Mn increased linearly with higher efficiency values in AP–BL than in BL–AP direction, however, the uptake of Mn revealed an asymptotic pattern within 120 min. In experiment 2, the kinetics of Mn transport in AP– X. Li J. Xie L. Lu L. Zhang L. Zhang X. Luo (&) Mineral Nutrition Research Division, Institute of Animal Science, Chinese Academy of Agricultural Sciences (CAAS), No. 2 Yuanmingyuan West Road, Haidian District, Beijing 100193, People’s Republic of China e-mail: [email protected] X. Li College of Animal Science and Technology, Henan University of Science and Technology, Luoyang 471003, People’s Republic of China Y. Zou Q. Wang S. Li (&) Key Laboratory of Preventive Veterinary Medicine in Hebei Province, Department of Animal Science, Hebei Normal University of Science and Technology, Qinhuangdao 066004, People’s Republic of China e-mail: [email protected]
BL was determined with media containing Mn concentrations from 0 to 2,500 lmol/L at 40 and 120 min, respectively, and mRNA levels of divalent metal transporter 1 (DMT1) and ferroportin (FPN1) were determined in Caco-2 cells treated with the medium containing 0 or 800 lmol/L of Mn for 120 min. The kinetics of Mn transport showed a carrier-mediated process when Mn concentrations were lower than 1,000 lmol/L and a linear increment when Mn concentrations exceeded 1,000 lmol/L at either 40 or 120 min. Mn treatment decreased (P \ 0.01) DMT1 mRNA level and increased (P \ 0.01) FPN1 mRNA level. The results from the present study suggested that Mn transport in AP–BL fit both carriermediated saturable and non-saturable diffusion processes, and Mn transport carriers DMT1 and FPN1 mediate the apical uptake and basolateral exit of Mn in Caco-2 cells. Keywords Manganese Transport Divalent metal transporter 1 Ferroportin Caco-2 cell
Introduction Manganese (Mn) is an essential metal found in various biological tissues, and it is necessary for normal functions of physiological processes including amino acid, lipid, protein and carbohydrate metabolism (Erikson et al. 2005). Manganese also plays an essential role in immune system functioning,
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Biometals
regulation of cellular energy, growth of bone and connective tissue and blood clotting (Eriks
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