活化的乙醛脱氢酶2通过调节双孔钾离子通道TASK-1减轻糖尿病大鼠心肌损伤

Activation of aldehyde dehydrogenase 2 attenuates myocardial injury in diabetic rats by regulating two-pore potassium channel TASK-1

目的:观察激活乙醛脱氢酶2(aldehyde dehydrogenase 2,ALDH2)对糖尿病心肌损伤中双孔钾离子通道TASK-1的影响及其对糖尿病心肌损伤的保护作用。方法:将雄性SD大鼠随机分为正常组(N组)、糖尿病4周组(DM4W组)、糖尿病8周组(DM8W组),糖尿病8周组+低浓度乙醇干预组(DM8W+Et OH组)。采用腹腔单次注射链脲佐菌素(55mg/kg)复制糖尿病大鼠模型。行心脏超声测定大鼠心功能,ELISA法检测心肌组织羟脯氨酸含量,HE染色观察心肌组织结构,PAS染色观察心肌组织阳性物质沉积情况,Western印迹检测心肌组织TASK-1蛋白表达情况,膜片钳记录心室肌细胞复极30%和90%时的动作电位时程(APD30,APD90)和双孔钾离子通道TASK-1电流,同时根据该钾通道对酸碱敏感的电生理特性判断是否为双孔钾离子通道TASK-1电流。结果:与N组相比,DM4W组和DM8W组大鼠左室舒张末期内径(end-diastole left ventricular diameter,LVIDd)及左室收缩末期内径(end-systolic left ventricular diameter,LVIDs)、羟脯氨酸含量、TASK-1蛋白表达增加,左室内径缩短率(leftventricular fractional shortening,LVFS)和射血分数(leftventricular ejection fraction,LVEF)均下降,APD30和APD90延长,TASK-1电流减少(P<0.01);HE染色结果显示DM4W组和DM8W组大鼠心肌细胞及纤维排列紊乱,心肌细胞肥大,心肌间隙增宽;PAS染色显示DM4W组和DM8W组大鼠心肌组织阳性物质沉积增加。与DM4W组相比,DM8W组大鼠上述指标的变化更加明显(P<0.05或P<0.01)。与DM8W相比,DM8W+Et OH组左室心功能指标LVIDd,LVIDs,LVFS,LVEF有所恢复,羟脯氨酸含量和TASK-1蛋白表达减少,TASK-1电流增加,APD30和APD90缩短(均P<0.01);HE染色显示心肌细胞损伤较前减轻,PAS染色显示大鼠心肌组织阳性物质沉积减少。结论:ALDH2被低浓度的乙醇激活后可减轻糖尿病所致的心肌损伤及纤维化,其机制可能与其调节双孔钾离子通道TASK-1蛋白表达和TASK-1电流有关。

Objective:To investigate the effect of activating aldehyde dehydrogenase 2(ALDH2)on TASK-1 two-pore potassium channel in myocardial injury of diabetic rats.Methods:Diabetic rats were induced by intraperitoneal injection of streptozotocin(55 mg/kg).The diabetic rats were divided into 4 groups:normal group,diabetes at 4th week(DM4W)group,diabetes at 8th week(DM8W)group,and diabetes at 8th week+low concentration of ethanol intervention(DM8W+EtOH)group.The cardiac function of rats was determined by cardiac ultrasonography.The content of hydroxyproline was detected by ELISA.The appearance of myocardial morphous and positive material were observed by HE and PAS staining.The protein expression of TASK-1 was detected by Western blot.Whole-cell patch clamp technique was used to record the action potential duration at 30%and 90%repolarization(APD30,APD90)and twopore potassium channel TASK-1 current in rat ventricular myocytes.Meanwhile,according to the sensitive electrophysiological characteristics of the potassium channel to acid and base,whether it is two-port potassium channel TASK-1current can be determined.Results:Compared with the N group,end-diastole left ventricular diameter(LVIDd),endsystolic left ventricular diameter(LVIDs),hydroxyproline content,TASK-1 protein expression increased,APD30 and APD90 extend,left ventricular fractional shortening(LVFS)and left ventricular ejection fraction(LVEF),and TASK-1 current decreased(all P<0.01)in the DM4W group and the DM8W group.HE staining showed that myocardial cell and fiber arrangement disorder,myocyte hypertrophy,myocardial widened and PAS staining reveals that positive material increased in the DM4W group and the DM8W group.Compared with the DM4W group,these changs are more obvious in DM8W rats(P<0.01 or P<0.05).Compared with the DM8W group,in the DM8W+EtOH group,the left ventricular function was restored,the hydroxyproline content and expression of TASK-1 protein were decreased,the TASK-1 current was increased,and APD30 and APD90 were shortened(all P<0.01).HE staining showed that myocardial cell injury was ameliorate and PAS staining showed decreased deposition of positive substances in the DM8W+EtOH group.Conclusion:Activation of aldehyde dehydrogenase 2 by low concentration of ethanol can reduce myocardial injury and fibrosis caused by diabetes,and its mechanism may be related to the changes of the two-por potassium channel TASK-1.