淀粉样β蛋白对猪脑微血管内皮细胞 ATP 结合盒转运蛋白表达和活性的影响。
Effect of amyloid beta on ATP-binding cassette transporter expression and activity in porcine brain microvascular endothelial cells.
机构信息
Division of Pharmacy & Optometry, University of Manchester, Manchester M13 9PT, UK.
Division of Pharmacy & Optometry, University of Manchester, Manchester M13 9PT, UK.
出版信息
Biochim Biophys Acta Gen Subj. 2018 Oct;1862(10):2314-2322. doi: 10.1016/j.bbagen.2018.07.021. Epub 2018 Jul 23.
BACKGROUND
Deposition of amyloid-β peptide (Aβ) within the brain is characteristic of Alzheimer's disease. Little is known of the effects of Aβ on blood-brain barrier (BBB) ATP-binding Cassette (ABC) efflux transporters which influence BBB permeability. The effects of Aβ on ABCB1, ABCC5 and ABCG2 activity and expression and pregnane X receptor (PXR) and constitutive androstane receptor (CAR) transcription factors expression were determined in primary porcine brain endothelial cells (PBECs).
METHODS
The effect of Aβ on transporter activity was determined by measurement of intracellular accumulation of the fluorescent probes calcein (ABCB1), GS-MF (ABCC5) and Hoechst 33342 (ABCG2). Expression of transporters and transcription factors was assessed by Western blotting.
RESULTS
Treatment of PBECs with Aβ significantly decreased activity of ABCB1 (Aβ at 10 μg/ml, 25 μg/ml and 50 μg/ml), ABCC5 (Aβ at 25 μg/ml and 50 μg/ml) and ABCG2 (Aβ at 10 μg/ml, 25 μg/ml and 50 μg/ml). Aβ also significantly decreased expression of ABCB1 (p < 0.05 at 25 μg/ml and 50 μg/ml), ABCG2 (p < 0.05 at 25 μg/ml and p ≤ 0.001 at 50 μg/ml), ABCC5 (p < 0.05 at 25 μg/ml and 50 μg/ml), PXR (p < 0.05 at 10 μg/ml, 25 μg/ml and 50 μg/ml Aβ) and CAR (p < 0.05 at 25 μg/ml and 50 μg/ml Aβ).
CONCLUSION
Aβ inhibits multiple ABC transporters and PXR and CAR in PBECs.
GENERAL SIGNIFICANCE
Aβ reduces ABC transporter activity and expression in BBB endothelial cells and has the potential to influence BBB permeability characteristics.
背景
淀粉样蛋白-β肽 (Aβ) 在大脑中的沉积是阿尔茨海默病的特征。目前人们对 Aβ 对血脑屏障 (BBB) ATP 结合盒 (ABC) 外排转运体的影响知之甚少，而这些转运体影响 BBB 的通透性。本研究旨在确定 Aβ 对原代猪脑内皮细胞 (PBEC) 中 ABCB1、ABCC5 和 ABCG2 活性和表达以及孕烷 X 受体 (PXR) 和组成型雄烷受体 (CAR) 转录因子表达的影响。
方法
通过测量荧光探针 calcein（ABCB1）、GS-MF（ABCC5）和 Hoechst 33342（ABCG2）在细胞内的积累来测定 Aβ 对转运体活性的影响。通过 Western blot 测定转运体和转录因子的表达。
结果
Aβ 处理 PBEC 可显著降低 ABCB1（Aβ 10μg/ml、25μg/ml 和 50μg/ml）、ABCC5（Aβ 25μg/ml 和 50μg/ml）和 ABCG2（Aβ 10μg/ml、25μg/ml 和 50μg/ml）的活性。Aβ 还显著降低了 ABCB1（p<0.05 在 25μg/ml 和 50μg/ml）、ABCG2（p<0.05 在 25μg/ml 和 p≤0.001 在 50μg/ml）、ABCC5（p<0.05 在 25μg/ml 和 50μg/ml）、PXR（p<0.05 在 10μg/ml、25μg/ml 和 50μg/ml Aβ）和 CAR（p<0.05 在 25μg/ml 和 50μg/ml Aβ）的表达。
结论
Aβ 抑制 PBEC 中的多种 ABC 转运体以及 PXR 和 CAR。
一般意义
Aβ 降低了 BBB 内皮细胞中 ABC 转运体的活性和表达，有可能影响 BBB 的通透性特征。
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