University of Rochester, Rochester, New York; and.
Am J Physiol Regul Integr Comp Physiol. 2014 Mar 1;306(5):R291-303. doi: 10.1152/ajpregu.00252.2013. Epub 2014 Jan 8.
The choroid plexus epithelium forms the blood-cerebrospinal fluid barrier and accumulates essential minerals and heavy metals. Choroid plexus is cited as being a "sink" for heavy metals and excess minerals, serving to minimize accumulation of these potentially toxic agents in the brain. An understanding of how low doses of contaminant metals might alter transport of other solutes in the choroid plexus is limited. Using primary cultures of epithelial cells isolated from neonatal rat choroid plexus, our objective was to characterize modulation of apical uptake of the model organic cation choline elicited by low concentrations of the contaminant metal cadmium (CdCl₂). At 50-1,000 nM, cadmium did not directly decrease or increase 30-min apical uptake of 10 μM [(3)H]choline. However, extended exposure to 250-500 nM cadmium increased [(3)H]choline uptake by as much as 75% without marked cytotoxicity. In addition, cadmium induced heat shock protein 70 and heme oxygenase-1 protein expression and markedly induced metallothionein gene expression. The antioxidant N-acetylcysteine attenuated stimulation of choline uptake and induction of stress proteins. Conversely, an inhibitor of glutathione synthesis l-buthionine-sulfoximine (BSO) enhanced stimulation of choline uptake and induction of stress proteins. Cadmium also activated ERK1/2 MAP kinase. The MEK1 inhibitor PD98059 diminished ERK1/2 activation and attenuated stimulation of choline uptake. Furthermore, inhibition of ERK1/2 activation abated stimulation of choline uptake in cells exposed to cadmium with BSO. These data indicate that in the choroid plexus, exposure to low concentrations of cadmium may induce oxidative stress and consequently stimulate apical choline transport through activation of ERK1/2 MAP kinase.
脉络丛上皮形成血脑屏障,并积累必需的矿物质和重金属。脉络丛被认为是重金属和多余矿物质的“汇”,有助于减少这些潜在有毒物质在大脑中的积累。对于低剂量污染物金属如何改变脉络丛中其他溶质的转运,我们的理解有限。本研究使用从新生大鼠脉络丛分离的上皮细胞原代培养物,旨在研究污染物金属镉(CdCl₂)对模型有机阳离子胆碱的顶端摄取的调节作用。在 50-1000 nM 浓度下,镉不会直接减少或增加 30 分钟内 10 μM [(3)H]胆碱的顶端摄取。然而,延长暴露于 250-500 nM 镉可使 [(3)H]胆碱摄取增加多达 75%,而无明显细胞毒性。此外,镉诱导热休克蛋白 70 和血红素加氧酶-1 蛋白表达,并显著诱导金属硫蛋白基因表达。抗氧化剂 N-乙酰半胱氨酸减弱了对胆碱摄取的刺激和应激蛋白的诱导。相反,谷胱甘肽合成抑制剂 l-丁硫氨酸亚砜(BSO)增强了对胆碱摄取的刺激和应激蛋白的诱导。镉还激活了 ERK1/2 MAP 激酶。MEK1 抑制剂 PD98059 减弱了 ERK1/2 的激活,并减弱了对胆碱摄取的刺激。此外,在暴露于 BSO 的镉的细胞中抑制 ERK1/2 的激活可减弱对胆碱摄取的刺激。这些数据表明,在脉络丛中,暴露于低浓度的镉可能会通过激活 ERK1/2 MAP 激酶诱导氧化应激,从而刺激顶端的胆碱转运。