UCP2在硫酸吲哚酚诱导心肌细胞肥大中的作用  

Effects of uncoupling protein 2 on cardiomyocyte hypertrophy induced by indoxyl sulfate

作　　者：徐新丽[1] 杨可[1] 管旭[1] 肖堂利[1] 何婷[1] 余彦琳 刘亮[1] 赵景宏[1] 

机构地区：[1]第三军医大学新桥医院肾内科,重庆400037

出　　处：《第三军医大学学报》2014年第22期2277-2282,共6页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81270290;30700316)~~

摘　　要：目的探讨解耦联蛋白2(uncoupling protein 2,UCP2)在硫酸吲哚酚(indoxyl sulfate,IS)诱导心肌细胞肥大中的作用和机制。方法分离培养新生Sprague-Dawley大鼠心肌细胞,0、100、250、500、1 000μmol/L IS孵育心肌细胞1 h或500μmol/L IS孵育心肌细胞0、15、30、45、60 min后,Western blot检测心肌细胞UCP2蛋白表达变化。UCP2慢病毒转染心肌细胞后,通过α-actinin免疫荧光染色观察细胞大小,3H-亮氨酸掺入法评价蛋白合成速率,Real-time PCR检测肥大相关基因ANF、BNP和β-MHC mRNA的表达,同时利用MitosoxTM探针检测活性氧(reactive oxygen species,ROS)生成情况。结果与对照组相比,IS处理组心肌细胞的UCP2蛋白水平明显下降(P<0.05);与IS处理组相比,UCP2过表达可以明显抑制IS诱导的心肌细胞ROS生成、体积增大、3H-亮氨酸摄入增加及肥大相关基因(ANF、BNP和β-MHC mRNA)的表达增高,差异有统计学意义(P<0.05)。结论 IS诱导心肌细胞肥大可能与其下调UCP2表达及促进氧化应激有关,而UCP2过表达或许可以通过抑制氧化应激拮抗IS诱导的心肌细胞肥大。Objective To investigate the effects of uncoupling protein 2（ UCP2） on cardiomyocyte hypertrophy induced by indoxyl sulfate（ IS）. Methods Neonatal rat cardiomyocytes were separately treated with IS of 0,100,250,500,and 1 000 μmol / L for 1 h or with 500 μmol / L IS for 0,15,30,45,and60 min. Then UCP2 expression was tested with Western blotting. After neonatal rat cardiomyocytes were infected with lentivirus,we observed the expression of UCP2 by Western blotting and detected reactive oxygen species（ ROS） production by Mitosox^TM. Cell size was tested by immunofluorescent staining of α-actinin.ANF,BNP and β-MHC mRNA and [^3H] leucine incorporation were examined as indicators of cardiomyocyte hypertrophy. Results Compared with the control group,IS treatment reduced the expression of UCP2 in a dose-and time-dependent manner（ P〈0. 05）. UCP2 overexpression significantly suppressed ROS generation,inhibited the increase of cell volume and [^3H] leucine incorporation,and reduced the mRNA expression of ANF,BNP,and β-MHC induced by IS（ P〈0. 05）. Conclusion IS may induce cardiomyocyte hypertrophy by down-regulating UCP2 expression and enhancing oxidative stress. UCP2 overexpression may antagonize ISinduced cardiomyocyte hypertrophy by inhibiting oxidative stress.

关 键 词：硫酸吲哚酚 心肌细胞肥大 解偶联蛋白2 机制 

分 类 号：R341[医药卫生—基础医学] R363.21