Suppression of Netrin-1 attenuates angiotension II-induced cardiac remodeling through the PKC/MAPK signaling pathway.
Suppression of Netrin-1 attenuates angiotension II-induced cardiac remodeling through the PKC/MAPK signaling pathway.
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DOI：
10.1016/j.biopha.2020.110495
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发表时间：
2020-07
期刊：
影响因子：
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通讯作者：
Gaojun Wu;Zhengxian Wang;P. Shan;Shanjun Huang;Shuang Lin;Weijian Huang;Zhouqing Huang
中科院分区：
文献类型：
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作者：
Gaojun Wu;Zhengxian Wang;P. Shan;Shanjun Huang;Shuang Lin;Weijian Huang;Zhouqing Huang
背景血管紧张素II（angiotensin II，Ang II）引起的心肌重构是心力衰竭的重要病理过程。Netrin-1是一种轴突导向因子，已被证明参与非神经元组织中的炎症反应、肿瘤发生和血管生成。然而，Netrin-1在心脏重塑中的作用尚未完全阐明。MethodsThe大鼠心肌细胞系H9 c2和原代新生大鼠心肌细胞用Ang II处理。用siRNA转染细胞以沉默Netrin-1表达。实时定量聚合酶链反应和蛋白质印迹法检测心肌细胞纤维化、凋亡和肥大的标志物。用Annexin V-EGFP/PI细胞凋亡检测试剂盒检测血管紧张素II诱导的H9 c2细胞和新生大鼠心肌细胞的凋亡水平。缬沙坦阻断AT 1 R可降低Netrin-1表达。重要的是，Netrin-1 siRNA的应用显著减轻了由Ang II诱导的心肌肥大、纤维化（由Myhc、胶原I和TGF-β反映）和凋亡（由Caspase 3、Bax和Bcl-2水平反映）的程度。此外，Netrin-1的沉默显著降低了PKCα、JNK和P38的磷酸化。我们分别用PKCα、JNK和P38抑制剂LY 317615、SP 600125和SB 203580处理H9 c2细胞，从而显著降低心肌肥厚、纤维化和凋亡。结论Ang II通过上调Netrin-1和激活AT 1 R/PKCα/MAPK（JNK，P38）通路，引起心肌肥厚、纤维化和凋亡。抑制Netrin-1可通过抑制PKCα/MAPK（JNK和P38）信号通路减轻Ang II诱导的心脏重构。因此，Netrin-1可能是Ang II介导的心脏重塑的一个新的治疗靶点。
BackgroundMyocardial remodeling caused by angiotensin II (Ang II) is essential for the pathological process of heart failure. Netrin-1, which is an axonal guidance cue, has been shown to be involved in the inflammatory response, tumorigenesis, and angiogenesis in non-neuronal tissues. However, the role of Netrin-1 in cardiac remodeling has not been fully elucidated.MethodsThe rat cardiomyocyte cell line H9c2 and primary neonatal rat cardiomyocytes were treated with Ang II. Cells were transfected with siRNA to silence Netrin-1 expression. Real-time polymerase chain reaction and Western blot analysis were used to detect the markers for fibrosis, apoptosis, and hypertrophy in cardiomyocytes. An Annexin V-EGFP/PI cell apoptosis detection kit was used to measure the level of apoptosis caused by angiotensin II.ResultsWe found that Netrin-1 expression was upregulated in the H9c2 cells and the neonatal rat cardiomyocytes stimulated by Ang II. The increased Netrin-1 expression was decreased by valsartan to block AT1R. Importantly, the application of Netrin-1 siRNA significantly alleviated the degrees of myocardial hypertrophy, fibrosis (reflected by Myhc, collagen I, and TGF-β) and apoptosis (reflected by the level of Caspase 3, Bax, and Bcl-2) induced by Ang II. In addition, the silencing of Netrin-1 substantially decreased the phosphorylation of PKCα, JNK, and P38. We treated H9c2 cells with LY317615, SP600125, and SB203580, inhibitors of PKCα, JNK, and P38, respectively, thereby resulting in a substantial decrease in hypertrophy, fibrosis, and apoptosis.ConclusionsAng II produces cardiac hypertrophy, fibrosis, and apoptosis through the upregulation of Netrin-1 and the activation of the AT1R/PKCα/MAPK (JNK, P38) pathway. Suppression of Netrin-1 can relieve Ang II-induced cardiac remodeling via inhibition of the PKCα/MAPK (JNK and P38) signaling pathway. Thus, Netrin-1 may be a novel therapeutic target for Ang II-mediated cardiac remodeling.