硫酸吲哚酚诱导心肌细胞肥大化的研究-第三军医大学学报.DOC

硫酸吲哚酚诱导心肌细胞肥大化的研究 杨 可，孙 蕾，管 旭，肖堂利，徐新丽，何 婷，赵景宏 （400037 重庆，第三军医大学新桥医院肾内科indoxyl sulfate，IS100、250、500、1000μmol/L2,7-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小时后，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小时后， 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 （P0.001），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（P0.001）。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。 [关键词] 硫酸吲哚酚；心肌细胞；肥大；氧化应激 [中图法分类号] [文献标志码] A Study of cardiomyocyte hypertrophy induced by indoxyl sulfate Yang Ke, Sun Lei, Guan Xu, Xiao Tangli, Xu Xinli, He Ting, Zhao Jinghong （Department of Nephrology, Hospital of Third Military Medical University, Chongqing, 400037, China） [Abstract] Objective To investigate the function and mechanism of Indoxyl Sulfate (IS) in inducing cardiac hypertrophy. Methods ardiac myocytes (NRCM) were isolated and treated with IS (100, 250, 500 and 1000μmol/L) for 3h before using fluorescence probe 2,7-dichlorofluorescin dictate (DCFH-DA) to detect the variation of production of intracellular reactive oxygen species (ROS). NRCM were incubated with IS (500μmol/L) for 24h, and then real-time PCR was applied to determine the changes of gene expression level of atrial natriuretic peptide (ANF), B-type natriuretic peptide (BNP) and beta-myosin heavy chain (β-MHC) mRNA. The cardiomyocytes were treated with IS (500μmol/L) for 48h, 3H-leucine incorporation assay and immunocytochemical technique were adopted to measure protein synthesis rate and to mark α-Smooth muscle actin (α-actinin) for observing morphology of NRCM respectively. Results IS caused a dose dependently increase of ROS synthesis (P0.001), and the expression levels of ANF, BNP and β-MHC mRNA as determined by real-time PCR were enhanced as well. IS (500μmol /L)