Nox4 and soluble epoxide hydrolase synergistically mediate homocysteine-induced inflammation in vascular smooth muscle cells
Nox4 and soluble epoxide hydrolase synergistically mediate homocysteine-induced inflammation in vascular smooth muscle cells
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Nox4和可溶性环氧化物水解酶协同介导同型半胱氨酸诱导的血管平滑肌细胞炎症
DOI：
10.1016/j.vph.2019.01.001
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发表时间：
2019-09-01
影响因子：
4
通讯作者：
Huang, Lan
中科院分区：
文献类型：
--
作者：
Liu, Xi;Qin, Zhexue;Huang, Lan
背景：高同型半胱氨酸血症导致血管平滑肌细胞（VSMC）炎症反应。同时，Nox 4依赖的活性氧（ROS）信号和可溶性环氧化物水解酶（sEH）/环氧二十碳三烯酸（ESTs）均参与血管炎症。在此，我们假设，Nox 4和可溶性环氧化物水解酶交叉调节同型半胱氨酸诱导的VSMC inflammation.Methods和结果：在培养的VSMC中，炎症因子VCAM 1和ICAM 1的表达通过实时PCR和Western印迹法测定，而上清液MCP 1通过ELISA测定。在用50 μ M同型半胱氨酸刺激VSMC后，我们观察到VCAM 1和ICAM 1 mRNA水平分别增加了1.15和1.0倍。用100 μ M处理培养的VSMCs的上清液中MCP 1水平增加到1.76倍。正如预期的那样，同型半胱氨酸诱导Nox 4表达和Nox 4依赖的ROS产生。在同型半胱氨酸存在下，sEH表达也以剂量依赖性方式上调。此外，我们用siRNA敲除了Nox 4。敲低Nox 4可降低ROS的产生和同型半胱氨酸诱导的sEH表达。用腺病毒过表达Nox 4刺激sEH表达。类似地，敲低或化学抑制sEH减弱了同型半胱氨酸对Nox 4的上调。在体内，在同型半胱氨酸喂养的小鼠，伴随着上调的Nox 4和sEH与增加VCAM 1和ICAM 1的表达在主动脉wall.Conclusions：在VSMCs同型半胱氨酸诱导的炎症反应伴随着Nox 4和sEH上调。Nox 4和可溶性环氧化物水解酶协同促进同型半胱氨酸诱导的炎症。
Background: Hyperhomocysteinemia leads to a vascular smooth muscle cell (VSMC) inflammatory response. Meanwhile, Nox4 dependent reactive oxygen species (ROS) signaling and soluble epoxide hydrolase (sEH)/epoxyeicosatrienoic acids (EETs) are both involved in vascular inflammation. Herein, we hypothesized that Nox4 and soluble epoxide hydrolase cross regulated during homocysteine-induced VSMC inflammation.Methods and results: In cultured VSMCs, the expression of the inflammatory factors VCAM1 and ICAM1 was measured by real-time PCR and Western blotting, while supernatant MCP1 was measured by ELISA. Upon VSMC stimulation with 50 mu M homocysteine, we observed the VCAM1 and ICAM1 mRNA levels were increased by 1.15 and 1.0 folds, respectively. The MCP1 levels in the supernatant of cultured VSMCs treated with 100 mu M increased to 1.76 folds. As expected, homocysteine induced Nox4 expression and Nox4-dependent ROS generation. The sEH expression was also upregulated in the presence of homocysteine in a dose-dependent manner. Furthermore, we knocked down Nox4 with siRNA. Knockdown of Nox4 decreased ROS generation and homocysteine-induced sEH expression. Overexpression of Nox4 with an adenovirus stimulated sEH expression. Similarly, knockdown or chemical inhibition of sEH blunted the upregulation of Nox4 by homocysteine. In vivo, in homocysteine-fed mice, concomitant upregulation of Nox4 and sEH was associated with increased VCAM1 and ICAM1 expression in the aortic wall.Conclusions: The inflammatory response induced by homocysteine in VSMCs was accompanied by Nox4 and sEH upregulation. Nox4 and soluble epoxide hydrolase synergistically contribute to homocysteine-induced inflammation.