心肌细胞肿胀激活氯离子通道单通道特性.doc

心肌细胞肿胀激活氯离子通道单通道特性 　 作者：王军，徐洪涛，吴燕红，孙喜庆 【关键词】 心肌/细胞学 Single channel properties of swellingactivated chloride channel in cardiac myocytes 　　【Abstract】 AIM: To characterize the single channel properties of swellingactivated chloride channel in freshly isolated mouse cardiac myocytes. METHODS: The cardiac myocytes were isolated from mouse hearts and put in a hypotonic solution. Cellattached and insideout patch clamp methods were employed to recorll the swellingactivated Cl- channel current (ICl.swell). RESULTS: Under isotonic conditions, no single channel activities were observed in most patches. Hypotonic superfusioninduced cell swelling resulted in the appearance of active channel. Under symmetrical condition, the current induced by hypotonicity showed outward rectification and an unitary conductance of （38.1±2.5）pS. The open probability of the channel was 0.76±0.08 at +100 mV in insideout patch recordings. The reversal potential (-34.5±2.8) mV of the current was close to the calculated equilibrium potential for Cl- (ECl=-38.6 mV). When ［Cl-］ changed, the reversal potential (Erev) of the current also shifted. The Cl- channel blocker DIDS reversibly inhibited the channel activity when added to the bath solution. CONCLUSION: In mouse cardiac myocytes, swellingactivated Cl- single channels show outward rectification and sensitivity to disulfonic stilbene blockerDIDS. 　　【Keywords】 myocardium/cytology; swellingactivated chloride channel; patchclamp techniques 　　【摘要】 目的： 研究急性分离小鼠心肌细胞肿胀激活氯离子通道的单通道特征. 方法： 采用膜片钳单通道（细胞贴附式和内面向外式）方法，记录急性分离的小鼠心肌细胞暴露于低渗溶液时出现的肿胀激活氯通道电流(ICl.swell). 结果： 等渗状态下，用细胞贴附式记录，多数心肌细胞膜片无通道活动，用低渗溶液灌流细胞，在细胞容积增大的同时，出现单通道电流的激活. 该肿胀激活单通道氯电压在-100 mV~+100 mV均有激活. 在电压+100 mV时，其单通道电导为(38.1±2.5) pS，开放概率为0.76±0.08，电流电压曲线显示其反转电位(-34.5±2.8 ) mV, 接近氯离子平衡电位的理论值(ECl=-38.6 mV), 且反转电位随氯离子浓度的改变而改变. 氯通道阻断剂DIDS可逆性阻断该单通道电流. 结论： 小鼠心肌细胞肿胀激活氯通道的单通道电导为(38.1±2.5) pS，电流呈外向整流特征并对DIDS敏感. 　　【关键词】 心肌/细胞学；肿胀激活氯离子通道；膜片钳术 　　0引言 　　调节性细胞容积减小（regulatory volume decrease, RVD）指的是细胞暴露于低渗透压环境时，细胞膨胀、容