Regulation of GADD153 induced by mechanical stress in cardiomyocytes
Regulation of GADD153 induced by mechanical stress in cardiomyocytes
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DOI：
10.1111/j.1365-2362.2009.02193.x
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发表时间：
2009-11-01
影响因子：
5.5
通讯作者：
Shyu, K. G.
中科院分区：
文献类型：
--
作者：
Cheng, W. P.;Wang, B. W.;Shyu, K. G.
细胞凋亡调控基因GADD 153（Growth arrest and DNA damage-inducible gene 153）在内质网应激时表达增加。然而，GADD 153在机械应力下心肌细胞中的表达知之甚少。本研究旨在探讨机械应力诱导心肌细胞GADD 153表达和凋亡的调控机制。材料与方法成年SD大鼠行主动脉-腔静脉分流术，诱导容量超负荷。将生长在柔性膜基底上的新生大鼠心肌细胞在60个周期min-1下通过真空拉伸至最大伸长的20%。结果在分流组（n = 8）中，超声心动图测量的心室尺寸增加通过口服伴侣4-苯基丁酸（PBA）500 mg/kg（d-1）治疗3d而逆转至正常。与假手术组（n = 8）相比，分流组GADD 153蛋白和mRNA表达上调。PBA处理使GADD 153蛋白逆转至基线值。TUNEL法显示PBA可减少容量超负荷诱导的细胞凋亡。周期性牵张14 h后GADD 153蛋白和mRNA表达显著增加。在牵张前30 min加入c-jun N-末端激酶（JNK）抑制剂SP 600125、JNK小干扰RNA和肿瘤坏死因子-α（TNF-α）抗体，减少了GADD 153蛋白的诱导。牵张增强了GADD 153-Mut质粒、SP 600125和TNF-α抗体对牵张诱导的GADD 153启动子活性的抑制作用。GADD 153 siRNA、GADD 153-Mut质粒和PBA均能逆转牵张诱导的GADD 153介导的细胞凋亡。PBA处理逆转了机械应力诱导的心肌细胞GADD 153表达和细胞凋亡。
P>BackgroundGrowth arrest and DNA damage-inducible gene 153 (GADD153), an apoptosis regulated gene, increased during endoplasmic reticulum stress. However, the expression of GADD153 in cardiomyocytes under mechanical stress is little known. We aimed to investigate the regulation mechanism of GADD153 expression and apoptosis induced by mechanical stress in cardiomyocytes.Materials and methodsAorta-caval shunt was performed in adult Sprague-Dawley rats to induce volume overload. Rat neonatal cardiomyocytes grown on a flexible membrane base were stretched by vacuum to 20% of maximum elongation, at 60 cycles min-1.ResultsThe increased ventricular dimension measured using echocardiography in the shunt group (n = 8) was reversed to normal by treatment with chaperon 4-phenylbutyric acid (PBA) (n = 8) at 500 mg kg-1 day-1 orally for 3 days. GADD153 protein and mRNA were up-regulated in the shunt group when compared with sham group (n = 8). Treatment with PBA reversed the protein of GADD153 to the baseline values. The TUNEL assay showed that PBA reduced the apoptosis induced by volume overload. Cyclic stretch significantly increased GADD153 protein and mRNA expression after 14 h of stretch. Addition of c-jun N-terminal kinase (JNK) inhibitor SP600125, JNK small interfering RNA and tumour necrosis factor-alpha (TNF-alpha) antibody 30 min before stretch, reduced the induction of GADD153 protein. Stretch increased, while GADD153-Mut plasmid, SP600125 and TNF-alpha antibody abolished the GADD153 promoter activity induced by stretch. GADD153 mediated apoptosis induced by stretch was reversed by GADD153 siRNA, GADD153-Mut plasmid and PBA.ConclusionsMechanical stress enhanced apoptosis and GADD153 expression in cardiomyocytes. Treatment with PBA reversed both GADD153 expression and apoptosis induced by mechanical stress in cardiomyocytes.