蛋白激酶C结合蛋白1参与心力衰竭小鼠心脏去甲肾上腺素转运蛋白的表达下调  

Protein interacting with kinase Cα mediates the down-regulation of myocardial norepinephrine transporter expression in mice with adriamycin-induced congestive heart failure

作　　者：靳文英[1] 乔正国[1] 郑春华[1] 李素芳[1] 陈红[1] 

机构地区：[1]北京大学人民医院心脏中心,100044

出　　处：《中华心血管病杂志》2014年第3期219-224,共6页Chinese Journal of Cardiology

基　　金：国家自然科学基金（31000472）;国家科技重大专项＂重大新药创制＂项目（2012ZX09303019）

摘　　要：目的 观察去甲肾上腺素转运蛋白（NET）和蛋白激酶C结合蛋白1（PICK1）的相互作用,并检测慢性心力衰竭小鼠心脏NET和PICK1 mRNA和蛋白的表达变化.方法 （1）细胞实验：293T细胞分为4组,分别转染质粒NET（NET组）、GFP-PICK1（GFP-PICK1组）、NET＋ GFP-PICK1（NET＋ GFP-PICK1组）以及NET＋ GFP-PICK1 （KD-AA）[NET＋ GFP-PICK1 （KD-AA）组],48 h后行免疫荧光化学染色进行亚细胞定位检测.（2）动物实验：雄性C57BL/6J小鼠分为阿霉素组和正常对照组（对照组）,每组各20只小鼠,分别于腹腔注射阿霉素（2 mg·kg-1·次-1,累计用量22 mg/kg）和生理盐水.实验第10周末采用实时荧光定量聚合酶链反应和蛋白质免疫印迹法检测心脏NET、PICK1和肾上腺素能受体（β1-AR）的mRNA和蛋白表达水平.结果 （1）PICK1与NET结合并介导NET的亚细胞定位.（2）阿霉素组和对照组小鼠颈交感神经节中NET mRNA的表达量分别为0.59±0.15和0.64±0.18,差异无统计学意义（P＞0.05）.但阿霉素组PICK1 mRNA表达量明显低于对照组（0.73 ±0.13比1.18 ±0.15,P＜0.05）,且β1-AR的mRNA表达量亦显著低于对照组（0.62 ±0.12比1.00 ±0.11,P＜0.05）.（3）阿霉素组小鼠心脏NET蛋白表达量明显低于对照组（0.72±0.04比0.75 ±0.03,P＜0.05）,而酪氨酸羟化酶蛋白表达量则显著高于对照组（0.95±0.06比0.84 ±0.07,P＜0.01）.（4）阿霉素组和对照组小鼠的心脏交感神经纤维密度无明显差异.结论 慢性心力衰竭小鼠心脏NET、PICK1的mRNA和蛋白表达均下调,PICK1介导的NET亚细胞定位障碍可能是心力衰竭时NET功能障碍的机制之一.Objective To investigate the interaction between myocardial norepinephrine （NET） and protein interacting with kinase Ca （ PICK1 ）, and examine the myocardial expression pattern of NET and PICK1 in mice with adriamycin-induced congestive heart failure. Methods （1） Cellular experiments： 293T cells were transfected with NET, GFP-PICK1, NET ＋ GFP-PICK1 or NET ＋ GFP-PICK1 （ KD-AA）, respectively. Immunofluorescence staining was performed 48 h after the transfection. （2） Animal experiments ： 40 male C57BL/6J mice were divided into control group and adriamycin group （ intraperitoneal injection of 2 mg/kg adriamycin with a cumulative amount of 22 mg/kg）. The myocardial mRNA and protein expression level of NET, PICK1 and adrenergic receptor （~31-AR） were detected by real-time PCR and Western blot after 10 weeks. Results （ 1 ） PICK1 mediates the intracellular trafficking of NET. （ 2 ） Compared to controls, cardiac mRNA expression of NET remained unchanged, but PICK1 and 131-AR mRNA level were significantly reduced in the heart failure mice. （3） Myocardial NET protein expression level was significantly reduced, whereas tyrosine hydroxylase （TH） protein expression was significantly upregulated in heart failure mice. （4） The myocardial density of sympathetic nerve fibers remained unchanged in heart failure mice. Conclusions Cardiac expression of NET and PICK1 are down-regulated in heart failure mice. Reduced PICKl-mediated intracellular trafficking of NET may be involved in theimpairment of NET function in this congestive heart failure mice model.

关 键 词：心力衰竭 充血性 蛋白激酶C 交感神经系统 

分 类 号：R541.6[医药卫生—心血管疾病]