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目的探讨容量敏感外向整流性(VSOR)氯通道在H2O2介导系膜细胞凋亡中的作用和可能的机制。方法全细胞膜片钳技术用于检测VSOR氯电流。细胞凋亡通过吖啶橙／溴化乙锭荧光染色、电子显微镜、TUNEL染色和半胱氨酸天冬氨酸蛋白酶(caspase)-3活性确定。结果150μmol／L H2O2激活系膜细胞VSOR氯电流,H2O2激活的氯电流具有典型的VSOR氯电流电生理特性。包括:外向整流性、电压依赖性失活。对细胞外高渗透压敏感及被VSOR氯通道阻断剂抑制。VSOR氯通道阻断剂DIDS(100μmol／L),NPPB(10μmol／L)和尼氟灭酸(10μmol／L)明显抑制H2O2介导的系膜细胞凋亡。150μmol／L H2O2处理2 h内,细胞容量明显下降,但这种细胞容量下降被100μmol／L DIDS,10μmol／L NPPB和10μmol／L尼氟灭酸抑制。结论VSOR氯通道参与H2O2介导的系膜细胞凋亡,其机制与介导凋亡性容量下降有关。
Objective To investigate the role and possible mechanism of capacity-sensitive outward rectifier (VSOR) chloride channel in H2O2-mediated apoptosis of mesangial cells. Methods Whole cell patch clamp technique was used to detect VSOR chloride current. Apoptosis was confirmed by acridine orange / ethidium bromide staining, electron microscopy, TUNEL staining and caspase-3 activity. Results The VSOR chloride current was activated by 150μmol / L H2O2. The chlorine current activated by H2O2 showed typical electrophysiological properties of VSOR chloride current. Including: outward rectification, voltage-dependent inactivation. Sensitivity to extracellular osmolarity and inhibition by VSOR chloride channel blockers. VSOR chloride channel blockers DIDS (100μmol / L), NPPB (10μmol / L) and niflumic acid (10μmol / L) significantly inhibited H2O2-mediated apoptosis of mesangial cells. The cell volume decreased obviously with the treatment of 150μmol / L H 2 O 2 for 2 h. However, the decrease of cell volume was inhibited by 100μmol / L DIDS, 10μmol / L NPPB and 10μmol / L niflumic acid. Conclusion VSOR chloride channel is involved in H2O2-mediated apoptosis of mesangial cells, and its mechanism is related to the decrease of apoptotic capacity.