Inhibition of CPU0213, a Dual Endothelin Receptor Antagonist, on Apoptosis via Nox4-Dependent ROS in HK-2 Cells
Inhibition of CPU0213, a Dual Endothelin Receptor Antagonist, on Apoptosis via Nox4-Dependent ROS in HK-2 Cells
复制标题
CPU0213（一种双重内皮素受体拮抗剂）通过 HK-2 细胞中 Nox4 依赖性 ROS 抑制细胞凋亡。
DOI：
10.1159/000445615
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发表时间：
2016-01-01
影响因子：
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通讯作者：
Xu, Ming
中科院分区：
文献类型：
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作者：
Li, Qing;Li, Ji;Xu, Ming
背景/目标：我们以前的研究表明，一种新的内皮素受体拮抗剂CPU 0213有效地使糖尿病肾病的肾功能正常化。然而，介导CPU 0213的肾保护作用的分子机制仍然未知。方法和结果：本研究首次检测了CPU 0213对人肾小管上皮细胞（HK-2）凋亡的影响。结果表明，高糖可显著上调HK-2细胞Bax蛋白表达，下调Bcl-2蛋白表达，CPU 0213可逆转高糖对HK-2细胞Bax蛋白表达的影响。显示Annexin V-FITC结合的HK-2细胞的百分比被CPU 0213显著抑制，这证实了CPU 0213对凋亡的抑制作用。鉴于内皮素（ET）系统对氧化应激的调节，我们确定了氧化还原信号在CPU 0213调节细胞凋亡中的作用。结果表明，在HK-2细胞中，CPU 0213处理显著减弱了超氧化物（O-2（-中心点））的产生。我们进一步发现CPU 0213显著抑制Nox 4蛋白的表达，这种基因沉默模拟了CPU 0213在高糖刺激下对细胞凋亡的作用。最后，我们检测了CPU 0213对ET-1受体的作用，发现CPU 0213显著抑制高糖诱导的内皮素A和B受体的蛋白表达。结论：高糖诱导HK-2细胞凋亡时，NO_x_4依赖的O_2 ~-的产生是细胞凋亡的关键。内皮素受体拮抗剂CPU 0213通过产生Nox 4依赖的O-2（-中心点）具有抗凋亡作用，这说明CPU 0213在糖尿病肾病中具有肾保护作用。(C)2016作者（s）由S. Karger AG，巴塞尔
Background/Aims: Our previous studies have indicated that a novel endothelin receptor antagonist CPU0213 effectively normalized renal function in diabetic nephropathy. However, the molecular mechanisms mediating the nephroprotective role of CPU0213 remain unknown. Methods and Results: In the present study, we first detected the role of CPU0213 on apoptosis in human renal tubular epithelial cell (HK-2). It was shown that high glucose significantly increased the protein expression of Bax and decreased Bcl-2 protein in HK-2 cells, which was reversed by CPU0213. The percentage of HK-2 cells that showed Annexin V-FITC binding was markedly suppressed by CPU0213, which confirmed the inhibitory role of CPU0213 on apoptosis. Given the regulation of endothelin (ET) system to oxidative stress, we determined the role of redox signaling in the regulation of CPU0213 on apoptosis. It was demonstrated that the production of superoxide (O-2(-center dot)) was substantially attenuated by CPU0213 treatment in HK-2 cells. We further found that CPU0213 dramatically inhibited expression of Nox4 protein, which gene silencing mimicked the role of CPU0213 on the apoptosis under high glucose stimulation. We finally examined the role of CPU0213 on ET-1 receptors and found that high glucose-induced protein expression of endothelin A and B receptors was dramatically inhibited by CPU0213. Conclusion: Taken together, these results suggest that this Nox4-dependenet O2-production is critical for the apoptosis of HK-2 cells in high glucose. Endothelin receptor antagonist CPU0213 has an anti-apoptosis role through Nox4-dependent O-2(-center dot) production, which address the nephroprotective role of CPU0213 in diabetic nephropathy. (C) 2016 The Author(s) Published by S. Karger AG, Basel