硫酸吲哚酚诱导心肌细胞肥大的研究  

Cardiomyocyte hypertrophy induced by indoxyl sulfate in vitro

作　　者：杨可[1] 孙蕾[1] 管旭[1] 肖堂利[1] 徐新丽[1] 何婷[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》2013年第24期2630-2633,共4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：目的研究尿毒症毒素硫酸吲哚酚(indoxyl sulfate,IS)对心肌细胞肥大的影响及其可能的机制。方法不同浓度的IS(100、250、500、1 000μmol/L)孵育体外培养的新生大鼠心肌细胞(RNCM)3 h后,2,7-dichlorofluorescin dictate(DCFH-DA)荧光探针检测细胞内活性氧(ROS)水平。500μmol/L IS孵育心肌细胞24 h后,Real-time PCR检测心肌细胞肥大标志基因心房尿钠肽(atrial natriuretic peptide,ANF)、B型尿钠肽(B-type natriuretic peptide,BNP)及β肌球蛋白重链(beta-myosin heavy chain,β-MHC)的mRNA表达。500μmol/L IS孵育NRCM 48 h后,3H亮氨酸掺入法检测心肌细胞蛋白合成速率,α-actinin免疫荧光染色观察心肌细胞面积。结果 IS可以剂量依赖性的诱导心肌细胞ROS生成增多(P<0.05),500μmol/L IS孵育可以导致心肌细胞肥大标志基因ANF、BNP和β-MHC mRNA表达上调,并显著增加心肌细胞3H亮氨酸摄入量和细胞面积(P<0.05)。结论 IS能够诱导体外培养的新生大鼠心肌细胞肥大,其机制可能与诱导氧化应激增强有关。Objective To investigate the function and mechanism of indoxyl sulfate （IS） in inducing cardiac hypertrophy. Methods Rat neonatal cardiac myocytes （RNCMs） were isolated and treated with IS （100, 250, 500 and 1 000μmol/L） for 3 h before using fluorescence probe 2,7-dichlorofluorescin dictate （DCFH-DA） to detect the variation of production of intracellular reactive oxygen species （ROS）. RNCMs were incubated with IS （500 μmol/L） for 24 h, and then real-time PCR was applied to determine the expression of atrial natriuretic peptide （ANF）, B-type natriuretic peptide （BNP） and beta-myosin heavy chain （J3-MHC） mRNA. The cardiomyocytes were treated with IS （500 μmol/L） for 48 h, 3 H-leucine incorporation assay and immunocytochemical technique were adopted to measure protein synthesis rate and to mark a-smooth muscle actin （ct-actinin） to observe morphology of RNCMs separately. Results IS caused a dose-dependent increase of ROS synthesis （P 〈 0.05 ） , and the expression levels of ANF, BNP and β-MHC mRNA were enhanced as well. IS （500 μmol/L） also exerted its function in increasing protein synthesis rate （P 〈 0. 05 ） and cell area in RNCMs. Conclusion IS can induce hypertrophy in cardiomyocytes, probably through the enhancement of oxidative stress.

关 键 词：硫酸吲哚酚 心肌细胞 肥大 氧化应激 

分 类 号：R-332[医药卫生] R322.11