CAJ | 학술논문

用膜片钳、反义寡核苷酸、免疫荧光及激光共聚焦显微镜等技术,研究MDR1基因在牛睫状体色素上皮(pigmented ciliary epithelial,PCE)细胞容积激活性氯电流中的作用.PCE细胞表达MDR1基因产物-P糖蛋白(P-gp).反义MDR1寡核苷酸抑制MDR1基因的表达(P-gp免疫荧光减少93%),延缓容积激活性氯电流的出现(潜伏期延长109%),并导致激活率降低62%及电流峰值减小56%.而核酸转染剂阳离子脂质体和非配对性的寡核苷酸对电流没有显著性影响.上述观察结果表明,睫状体色素上皮细胞容积激活性氯电流与内源性MDR1 表达有关.
용막편겸、반의과핵감산、면역형광급격광공취초현미경등기술,연구MDR1기인재우첩상체색소상피(pigmented ciliary epithelial,PCE)세포용적격활성록전류중적작용.PCE세포표체MDR1기인산물-P당단백(P-gp).반의MDR1과핵감산억제MDR1기인적표체(P-gp면역형광감소93%),연완용적격활성록전류적출현(잠복기연장109%),병도치격활솔강저62%급전류봉치감소56%.이핵산전염제양리자지질체화비배대성적과핵감산대전류몰유현저성영향.상술관찰결과표명,첩상체색소상피세포용적격활성록전류여내원성MDR1 표체유관.
The role of multidrug resistance (MDR1) gene in the activation of volume-activated chloride currents in bovine pigmented ciliary epithelial (PCE) cells was investigated by the patch-clamp technique, the antisense approach, the immunofluorescent technique and the confocal microscopy. PCE cells express P-glycoprotein (P-gp, the product of MDR1 gene). An MDR1 antisense oligonucleotide suppressed MDR1 expression (93% reduction of P-gp immunofluorescence), delayed the activation of a volume-activated chloride current (latency prolonged by 109%), reduced the activation rate by 62% and decreased the peak value of the current by 56%. The transfection reagent lipofectin and the mismatch control oligonucleotide did not significantly affect the current. The data indicate that the volume-activated chloride current is associated with the endogenous expression of MDR1 gene in PCE cells.