Tachikawa Masanori, Toki Hidetoh, Tomi Masatoshi, Hosoya Ken-ichi
Department of Pharmaceutics, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama 930-0194, Japan.
Microvasc Res. 2008 Jan;75(1):68-72. doi: 10.1016/j.mvr.2007.05.002. Epub 2007 May 18.
The purpose of this study was to quantify gene expression levels of the ATP-binding cassette (ABC) transporter A and C subfamilies ABCA1-A9, and ABCC1-6/Mrp1-6, C10/Mrp7 in mouse retinal vascular endothelial cells (RVEC) using a combination of a magnetic isolation method for mouse RVEC and real-time quantitative PCR analysis. The transcript level of endothelial cell markers, such as CD31, Tie-2, claudin-5, occludin, ABCB1a/mdr1a, and ABCG2, were more than 20-fold higher than those in the non-RVEC fraction, suggesting that RVEC in the RVEC fraction are concentrated at least 20-fold compared with those of the non-RVEC fraction. In the ABCA1 to A9 families, the transcript level of ABCA3 and A9 in the RVEC fraction was 1.2- and 32-fold higher than that in the non-RVEC fraction. Although ABCA3 was expressed in both the RVEC and non-RVEC fractions, A9 is predominantly expressed in the RVEC fraction. In the ABCC1 to C6 and C10 families, the transcript level of ABCC3, C4, and C6 in the RVEC fraction was 27-, 251-, and 242-fold higher, respectively, than that in the non-RVEC fraction, suggesting that ABCC3, C4, and C6 are predominantly expressed in the RVEC. In conclusion, ABCA3, ABCA9, ABCC3, ABCC4, and ABCC6 mRNAs are predominantly expressed at the inner blood-retina barrier (inner BRB) and appear to play a major role in the efflux transport of their substrates at the inner BRB.
本研究的目的是结合小鼠视网膜血管内皮细胞(RVEC)的磁分离方法和实时定量PCR分析,对ATP结合盒(ABC)转运蛋白A和C亚家族ABCA1 - A9以及ABCC1 - 6/Mrp1 - 6、C10/Mrp7在小鼠视网膜血管内皮细胞中的基因表达水平进行定量。内皮细胞标志物如CD31、Tie - 2、claudin - 5、occludin、ABCB1a/mdr1a和ABCG2的转录水平比非RVEC组分中的转录水平高20倍以上,这表明与非RVEC组分相比,RVEC组分中的RVEC至少浓缩了20倍。在ABCA1至A9家族中,RVEC组分中ABCA3和A9的转录水平分别比非RVEC组分高1.2倍和32倍。虽然ABCA3在RVEC和非RVEC组分中均有表达,但A9主要在RVEC组分中表达。在ABCC1至C6和C10家族中,RVEC组分中ABCC3、C4和C6的转录水平分别比非RVEC组分高27倍、251倍和242倍,这表明ABCC3、C4和C6主要在RVEC中表达。总之,ABCA3、ABCA9、ABCC3、ABCC4和ABCC6 mRNA主要在内血视网膜屏障(内BRB)表达,并且似乎在内BRB的底物外排转运中起主要作用。