Wen Yueqiang, Chen Jieru, Liu Lingling, Guo Guanhua, Zeng Yinsi, Zhang Yujing, Zeng Zhili, Wang Zebin, Peng Xuan, Liang Jianbo
Department of Nephrology, The Second Affiliated Hospital, Guangzhou Medical University, Guangzhou, China.
Department of Intensive Care Unit, The Second Affiliated Hospital, Guangzhou Medical University, Guangzhou, China.
J Recept Signal Transduct Res. 2020 Apr;40(2):141-147. doi: 10.1080/10799893.2020.1719518. Epub 2020 Jan 31.
Angiotensin II (AngII) induced Calcineurin binding protein 1 (Cabin1) protein expression significantly increased during Renal tubular epithelial cells (RTEC) injury. However, the detailed function of Cabin1 protein in RTEC was not characterized well. In this study, we aimed to explore the downstream target of Cabin1 model. Rat kidney epithelial cells were cultured and stimulated with AngII. Electron microscopy was performed to observe mitochondrial morphology change. Immunofluorescence staining was detected to observe the distribution of cytoskeleton and Cabin1. Mitochondrial morphology change and protein expression were detected by electrical microscopy and western blot. AngII induced the disruption of cytoskeleton at 24 and 48 h. Western blot analysis showed AngII significantly induced the overexpression of Cabin1. AngII induced a great deal of small, long and irregular mitochondria in RTEC, aspect ratio which reflects the length-to-width ratio of mitochondria remarkably increased at 12 and 24 h. Knocking down Cabin1 aggravated mitochondrial morphological abnormality in AngII treated RTEC. In comparison with control, Cabin1, p53 and cyto C level were significantly increased in AngII treated cells, while SIRT1 level was obviously decreased. Knocked down Cabin1 plus AngII stimulated, SIRT1 was further decreased, while p53 and cyto C were significantly increased. Cabin1 involves in RTEC mitochondrial dysfunction through SIRT1/p53 pathway. Cabin1 may be used as a new marker for the mechanisms of RTEC injury.
血管紧张素II（AngII）诱导的钙调神经磷酸酶结合蛋白1（Cabin1）蛋白表达在肾小管上皮细胞（RTEC）损伤期间显著增加。然而，Cabin1蛋白在RTEC中的详细功能尚未得到很好的表征。在本研究中，我们旨在探索Cabin1的下游靶点模型。培养大鼠肾上皮细胞并用AngII刺激。进行电子显微镜观察线粒体形态变化。检测免疫荧光染色以观察细胞骨架和Cabin1的分布。通过电子显微镜和蛋白质印迹法检测线粒体形态变化和蛋白质表达。AngII在24小时和48小时诱导细胞骨架破坏。蛋白质印迹分析显示AngII显著诱导Cabin1的过表达。AngII在RTEC中诱导大量小的、长的和不规则的线粒体，反映线粒体长宽比的纵横比在12小时和24小时显著增加。敲低Cabin1会加重AngII处理的RTEC中的线粒体形态异常。与对照相比，AngII处理的细胞中Cabin1、p53和细胞色素C水平显著增加，而SIRT1水平明显降低。敲低Cabin1加AngII刺激后，SIRT1进一步降低，而p53和细胞色素C显著增加。Cabin1通过SIRT1/p53途径参与RTEC线粒体功能障碍。Cabin1可能作为RTEC损伤机制的新标志物。
