氯通道ClC-3反义寡核苷酸对过氧化氢诱导的大鼠主动脉平滑肌细胞凋亡的影响

目的研究ClC3反义寡核苷酸对H2O2诱导的大鼠主动脉平滑肌细胞凋亡的影响。方法蛋白免疫印迹法检测ClC3蛋白表达;形态学方法、DNA琼脂糖电泳、MTT法和流式细胞仪观察和分析H2O2诱导的大鼠主动脉平滑肌细胞形态学改变、DNA断裂、细胞存活率和凋亡率及ClC3反义寡核苷酸转染对其影响。结果ClC3反义寡核苷酸转染抑制内源性ClC3蛋白表达后,可加重H2O2诱导大鼠主动脉平滑肌细胞形态学改变及DNA断裂,细胞凋亡率由52.8%±13.6%增至75.7%±5.8%(n=6,P<0.01),而细胞存活率由48.9%±4.3%进一步降低为31.3%±4.3%(n=6,P<0.01)。结论ClC3反义寡核苷酸转染促进H2O2诱导的大鼠主动脉平滑肌细胞凋亡。

Effects of ClC-3 Antisense Oligonucleotide on H2O2-Induced Apoptosis in Rat Aortic Vascular Smooth Muscle Cells

Aim To investigate the effects of ClC-3 antisense oligonucleotide on H2_O2-induced apoptosis in rat aortic vascular smooth muscle cells. Methods Western-blot was performed to detect the expression of ClC-3 protein. Morphological determination, DNA agarose gel electrophoresis, MTT assay and flow cytometry analysis were performed to measure the morphological changes, DNA ladder, viability and apoptotic rate and effects of ClC-3 antisense oligonucleotide on rat aortic smooth muscle cells. Results The antisense oligonucleotide specific to ClC-3 mRNA decreased the expression of ClC-3 protein and enhanced the apoptotic events induced by H₂O₂ including ultrastructural alteration and DNA ladder. The apoptotic cell death rate increased from 52.8%±13.6% to 75.7%±5.8% (n=6,p<0.01),and whereas the viability further decreased from 48.9%±4.3% to 31.3%±4.3 % (n=6,p<0.01) by ClC-3 antisense oligonucleotide transfection. Conclusion ClC-3 antisense oligonucleotide potentiated apoptosis induced by H₂O₂ in rat aortic smooth muscle cells.