Oberbauer R, Schreiner G F, Biber J, Murer H, Meyer T W
Department of Medicine, Palo Alto Veterans Administration Medical Center, CA 94303, USA.
Proc Natl Acad Sci U S A. 1996 May 14;93(10):4903-6. doi: 10.1073/pnas.93.10.4903.
A 20-mer phosphorothioate oligonucleotide (AS1) was designed to hybridize to the message for the rat kidney sodium phosphate cotransporter NaPi-2 close to the translation initiation site. Single intravenous doses of this oligonucleotide were given to rats maintained on a low phosphorus diet to increase NaPi-2 expression. At 3 days after oligonucleotide infusion, rats receiving 2.5 micromol of AS1 exhibited a reduction in renal NaPi-2 to cyclophilin mRNA ratio by 40% +/- 17%, and rats receiving 7.5 micromol of AS1 exhibited a reduction in NaPi-2 to cyclophilin mRNA ratio by 46% +/- 21%. Reversed-sequence AS1 was without effect. The higher dose of 7.5 micromol of AS1 also reduced the rate of phosphate uptake into renal brush border membrane vesicles and the expression of NaPi-2 protein detected by Western blotting in these vesicles. Reversed sequence AS1 was again without effect on these parameters. These results suggest that systemically infused oligonucleotides can exert antisense effects in the renal proximal tubule.
设计了一种20聚体硫代磷酸酯寡核苷酸（AS1），使其与大鼠肾钠磷共转运体NaPi-2的信使核糖核酸在靠近翻译起始位点处杂交。将该寡核苷酸单次静脉注射给维持低磷饮食以增加NaPi-2表达的大鼠。在寡核苷酸输注后3天，接受2.5微摩尔AS1的大鼠肾NaPi-2与亲环蛋白信使核糖核酸的比率降低了40%±17%，接受7.5微摩尔AS1的大鼠NaPi-2与亲环蛋白信使核糖核酸的比率降低了46%±21%。反向序列的AS1没有作用。7.5微摩尔的高剂量AS1也降低了磷酸盐摄取到肾刷状缘膜囊泡中的速率以及通过蛋白质印迹法在这些囊泡中检测到的NaPi-2蛋白的表达。反向序列的AS1再次对这些参数没有影响。这些结果表明，全身输注的寡核苷酸可在肾近端小管发挥反义作用。
