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 μmol/L of Mn (MnSO4·H2O). Both the uptake and transport of Mn from AP-BL or from BL-AP at different time-points 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-BL was determined with media containing Mn concentrations from 0 to 2,500 μmol/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 μmol/L of Mn for 120 min. The kinetics of Mn transport showed a carrier-mediated process when Mn concentrations were lower than 1,000 μmol/L and a linear increment when Mn concentrations exceeded 1,000 μmol/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 carrier-mediated 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.

译文

:进行了两个实验,以研究锰(Mn)在Caco-2细胞单层中的迁移动力学以及Mn(转运)载体在顶(AP)和基底外侧(BL)膜中的基因表达。在实验1中,用含146μmol/ L Mn(MnSO4·H2O)的培养基处理细胞。评估了在不同时间点从AP-BL或BL-AP吸收和转运Mn的情况,以确定Mn转运动力学的最佳时间。在AP-BL中,Mn的运输以比BL-AP方向高的效率值线性增加,但是,Mn的吸收显示120分钟内的渐近模式。在实验2中,使用含有Mn浓度从0到2500μmol/ L的培养基分别在40和120分钟测定AP-BL中Mn转运的动力学,以及二价金属转运蛋白1(DMT1)和铁转运蛋白(FPN1)的mRNA水平。在用含0或800μmol/ L Mn的培养基处理120分钟的Caco-2细胞中测定)。当Mn浓度低于1,000μmol/ L时,Mn迁移的动力学表现出一个由载体介导的过程,而当Mn浓度超过1,000μmol/ L时,在40或120 min时,其迁移呈线性增加。锰处理降低(P <0.01)DMT1 mRNA水平和增加(P <0.01)FPN1 mRNA水平。本研究的结果表明,AP-BL中的Mn转运既符合载体介导的饱和和非饱和扩散过程,而Mn转运载体DMT1和FPN1介导Caco-2细胞中Mn的根尖吸收和基底外侧出口。

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