反义多重耐药性基因抑制眼睫状体非色素上皮细胞容积激活性氯电流.docVIP

???? 反义多重耐药性基因抑制眼睫状体非色素?上皮细胞容积激活性氯电流 ???? ????［摘　要］　目的：研究多重耐药性(MDR1)基因及其产物P糖蛋白(P-gp)与容积激活性Cl-电流的关系。方法：用膜片钳全细胞记录技术记录牛眼睫状体非色素上皮(NPCE)细胞容积激活性Cl-电流，反义寡核苷酸阻抑细胞MDR1基因表达，在激光共聚焦显微镜下检测细胞P-gp免疫荧光。结果：P-gp免疫荧光与人源反义MDR1呈剂量依赖性减弱关系,容积激活性Cl-电流被人源反义MDR1特异性地部分阻抑，电流潜伏期延长，峰电流值减少，电流抑制率与人源反义MDR1呈现剂量依赖性增强关系,(r=0.99， P＜0.01)。 P-gp表达抑制率和容积激活性Cl-电流抑制率高度正相关(r=0.99， P＜0.01)。结论：MDR1基因及其产物P-gp参与NPCE细胞容积激活性Cl-电流的形成。　　［主题词］　离子通道； 寡核苷酸类，反义;抗药性;基因表达　　［中分类号］　Q 25　　　［文献标识码］　A［文章编号］　1000-4718(2000)07-0658-05 Multidrug-resistance antisense inhibits volume-activated chloridecurrent in non-pigmented ciliary epithelial cells CHEN Li-xin, Wang Li-wei(Department of Physiology, Guangdong Medical College, Zhanjiang 524023, China) 　　［Abstract］　AIM： To study the relationship between multidrug-resistance (MDR1) gene product P-glycoprotein (P-gp) and the volume-activated chloride current. METHODS:The volume-activated chloride current in bovine non-pigmented ciliary epithelial cells was recorded using a whole cell recording technique. An antisense technique was used to inhibit the expression of MDR1 gene. The immunofluorescence of P-gp was monitored with a real-time laser confocal microscope.RESULTS:P-gp immunofluorescence correlated negatively with the concentration of the human MDR1 antisense oligonucleotide. The antisense oligonucleotide inhibited the volume-activated chloride current specifically and partially. The latency of activation of the current increased and the peak current decreased. The percentage of inhibition of peak current correlated positively to the concentration of the antisense oligonucleotide(r＝0.99, P＜0.01) and to the reduction(%) of P-gp immunofluorescence(r=0.99,P＜0.01).CONCLUSION:P-gp, the product of MDR1 gene, plays an important role in the activation pathway of volume-activated chloride current in non-pigmented ciliary epithelial cells.　　［MeSH］　Ion channels; Oligonucleotides, antisense; Drug resistance; Gene expression 　　睫状体非色素上皮(non-pigmented ciliary epithelium, NPCE)细胞分泌房水机制与