钙/钙调素依赖性蛋白激酶Ⅱ抑制剂KN-93对兔心衰模型心室肌细胞晚钠电流的影响  

Effect of Ca^(2+)/calmodulin-dependent protein kinaseⅡinhibitor KN-93 on late sodium current in rabbits model of heart failure

作　　者：刘衍冬 颜素娟[2] 闫双冰 董泉彬 郑安财 李凡 裴兆辉 李菊香[2] 

机构地区：[1]南昌市第三医院心内二科 [2]南昌大学第二附属医院心内科 [3]南昌大学医学院,江西南昌330006

出　　处：《中国病理生理杂志》2017年第11期1964-1968,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81200132);江西省研究生创新专项资金资助项目(No.YC2014-S085)

摘　　要：目的:探讨异丙肾上腺素(isoproterenol,ISO)诱导兔心衰(heart failure,HF)时,兔心室肌细胞晚钠电流(late sodium current,I_(Na L))的变化及钙/钙调素依赖性蛋白激酶Ⅱ(Ca^(2+)/calmodulin-dependent protein kinaseⅡ,Ca MKⅡ)抑制剂KN-93对I_(Na L)的影响。方法:经兔耳缘静脉注射ISO(300μg·kg^(-1)·d^(-1),连续15 d)诱导兔HF模型;1月后行心脏彩超检查和HE染色观察心肌形态改变来评价HF模型;Western blot分析Na_V1.5、Ca MKⅡδ和磷酸化Ca MKⅡδ的蛋白水平;经Langendorff装置灌注消化生理盐水(normal saline,NS)组和HF组的兔心室肌细胞,采用全细胞膜片钳技术记录I_(Na L)。结果:与NS组相比,HF组兔心率增加(P<0.01),心室腔增大(P<0.05),心功能明显降低(P<0.01);心肌细胞排列紊乱,呈空泡样变性,细胞间质存在水肿,纤维组织增多。HF组的Na_V1.5、Ca MKⅡδ及磷酸化Ca MKⅡδ的蛋白水平与NS组相比均明显增加(P<0.01)。HF组的I_(Na L)与NS组相比明显增加(P<0.01);加入海葵毒素Ⅱ(sea anemone toxinⅡ,ATXⅡ)后,HF组及NS组I_(Na L)均明显增加,且HF组比NS组增加更明显(P<0.01);在ATXⅡ诱导电流增加稳定后,加入KN-93,NS组及HF组中I_(Na L)均明显减少(P<0.05)。结论:Ca MKⅡ抑制剂KN-93能抑制心衰增加的I_(Na L),这一作用可能与影响Ca MKⅡδ活性及Ca MKⅡδ参与I_(Na L)的调控有关。AIM：To investigate the change of late sodium current（INa L）and the effect of Ca2＋/calmodulindependent protein kinaseⅡ（Ca MKⅡ）inhibitor KN-93 on INa Lin the cardiomyocytes after isoproterenol-induced heart failure（HF）in rabbits.METHODS：The rabbit model of HF was induced by injecting isoproterenol（300μg·kg-1·d-1）for 15 d.One month later,all rabbits received by echocardiography and HE staining to observe the morphological changes of myocardium for evaluating the HF model.The protein expression of NaV1.5,Ca MKⅡδand phosphorylated Ca MKⅡδwas determined by Western blot.The ventricular myocytes were isolated from the rabbits of normal saline（NS）group and HF group by Langendorff perfusion,and the whole-cell patch-clamp technique was used to record INa L.RE-SULTS：Compared with NS group,the heart rate in HF group was increased（P〈0.01）,the ventricular cavity was enlarged（P〈0.05）,and the cardiac function was decreased（P〈0.01）.Compared with NS group,the cardiomyocytes in HF group arranged in disorder,vacuolar degeneration and myocardial interstitial edema were observed,and fibrous tissue increased.The protein levels of NaV1.5,Ca MKⅡδand phosphorylated Ca MKⅡδin HF group were higher than those in NS group（P〈0.01）.INa Lin HF group significantly increased compared with NS group（P〈0.01）.After adding sea anemone toxinⅡ（ATXⅡ）,the density of INa Lin HF group and NS group was significantly increased,but that in HF group increased more obviously than that in NS group（P〈0.01）.After ATXⅡhad induced stable current,we added KN-93 into NS group and HF group,and we found that the ATXⅡ-increased INa Lin NS group and HF group was significantly decreased（P〈0.05）.CONCLUSION：Ca MKⅡinhibitor KN-93 inhibits the increase in INa Lin HF rabbits,which may be related to the activity of Ca MKⅡδand the regulation of Ca MKⅡδon INa L.

关 键 词：钙/钙调素依赖性蛋白激酶Ⅱ 心力衰竭 晚钠电流 KN-93 

分 类 号：R541.6[医药卫生—心血管疾病]