High glucose activates Raw264.7 macrophages through RhoA kinase-mediated signaling pathway
High glucose activates Raw264.7 macrophages through RhoA kinase-mediated signaling pathway
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DOI：
10.1016/j.cellsig.2014.11.012
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发表时间：
2015-02-01
影响因子：
4.8
通讯作者：
Kao, Ying-Hsien
中科院分区：
文献类型：
--
作者：
Cheng, Cheng-I;Chen, Po-Han;Kao, Ying-Hsien
高血糖已被证明会加速动脉粥样硬化形成，这是一种由巨噬细胞激活引起的炎症过程。尽管高葡萄糖 (HG) 先前已被证明可增强巨噬细胞中的 ROCK 活性并增强其体外激活，但 ROCK 信号传导在 HG 介导的巨噬细胞激活中的作用仍不清楚。本研究旨在阐明 HG 介导的 ROCK 上调和巨噬细胞激活的潜在信号转导途径，包括 c-Jun 或 NF-kappa B 途径。使用巨噬细胞系 RAW264.7 研究 HG 对 RhoA/ROCK 活性和巨噬细胞功能的致动脉粥样硬化影响。暴露于 HG 显着诱导 RhoA 膜易位、RhoA 激酶活性和肌球蛋白结合亚基（RhoA 激酶底物）的磷酸化。巨噬细胞的行为，包括细胞增殖、粘附、迁移和 TNF-α 从头合成，也因 HG 暴露而增加。然而，羟基法舒地尔的药理学 ROCK 抑制作用减弱了 HG 增强的粘附和 TNF-α 的产生。 HG 刺激后，同时观察到 c-Jun 和转录因子 NF-κ B 的核转位。羟基法舒地尔的药理学 ROCK 抑制作用和 siRNA 介导的 ROCK1 或 ROCK2 基因沉默证实了巨噬细胞中 ROCK 依赖性 JNK 和 ERK 磷酸化，但未证实 NF-κ B 激活。此外，这两种干预措施都有效改善了模拟糖尿病条件下 HG 介导的巨噬细胞活化。这些发现表明，高血糖主要通过 ROCK/JNK 和 ROCK/ERK 途径激活巨噬细胞，从而导致更促炎的表型，并最终导致动脉粥样硬化形成。总之，ROCK 抑制可能成为糖尿病患者动脉粥样硬化治疗和预防的新治疗策略。 (C) 2014 爱思唯尔公司保留所有权利。
Hyperglycemia has been shown to accelerate atherogenesis, an inflammation process resulting from macrophage activation. Although high glucose (HG) was previously demonstrated to accentuate ROCK activity in macrophages and enhance their activation in vitro, the role of ROCK signaling in HG-mediated macrophage activation remains unclear. This study aimed to elucidate potential signal transduction pathways of HG-mediated ROCK upregulation and macrophage activation, including c-Jun or NF-kappa B pathways. A macrophage cell line, RAW264.7, was used to investigate the atherogenic effects of HG on RhoA/ROCK activity and macrophage functions. Exposure to HG significantly induced RhoA membrane translocation, RhoA-kinase activity, and phosphorylation of myosin-binding subunit, a RhoA-kinase substrate. Macrophage behaviors, including cell proliferation, adhesion, migration, and TNF-alpha de novo synthesis, were also increased by HG exposure. However, pharmacological ROCK inhibition by hydroxyfasudil attenuated the HG-enhanced adhesion and TNF-alpha production. Nuclear translocation of c-Jun and transcription factor NF-kappa B was simultaneously noted after HG stimulation. Pharmacological ROCK inhibition by hydroxyfasudil and siRNA-mediated ROCK1 or ROCK2 gene silencing confirmed the ROCK-dependent JNK and ERK phosphorylation, but not NF-kappa B activation in macrophages. In addition, both interventions effectively ameliorated the HG-mediated macrophage activation under the conditions mimicking diabetes. These findings suggest that hyperglycemia activates macrophages mainly through ROCK/JNK and ROCK/ERK pathways, which results in a more pro-inflammatory phenotype and eventually contributes to atherogenesis. In conclusion, ROCK inhibition might become a novel therapeutic strategy in atherosclerosis treatment and prevention in diabetic patients. (C) 2014 Elsevier Inc All rights reserved.