Homocysteine-induced oxidative stress upregulates chymase in mouse mastocytoma cells
Homocysteine-induced oxidative stress upregulates chymase in mouse mastocytoma cells
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DOI：
10.1038/hr.2009.205
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发表时间：
2010-02-01
影响因子：
5.4
通讯作者：
Urata, Hidenori
中科院分区：
文献类型：
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作者：
Fujimi, Kanta;Uehara, Yoshinari;Urata, Hidenori
过氧化氢(H_2O_2)、O-2(中心点-)和OH中心点等活性氧物种(ROS)参与了缺血/再灌注损伤、炎症和动脉粥样硬化的发病过程。我们以前的研究表明，血管紧张素II(Ang II)形成糜酶的增加可能参与了动脉粥样硬化的发展。然而，目前对凝乳酶表达的调控机制还不是很清楚。在这项研究中，我们测试了氧化应激是否上调了小鼠肥大细胞蛋白酶、小鼠肥大细胞蛋白酶(MMCP)-5或MMCP-4。我们还检测了这些蛋白在治疗后的表达和活性。培养的小鼠肥大细胞瘤细胞(MMC)具有糜酶依赖的Ang II形成活性，用过氧化氢和几种氨基硫醇加或不加抗氧化剂处理。用定量RT-PCR法检测MMCP-5和MMCP-4的表达水平，以血管紧张素转换酶I为底物，用高效液相色谱法测定凝乳酶依赖的血管紧张素Ⅱ形成活性。同型半胱氨酸(0.1~3mmoL L(-1))作用于MMC后，MMCP-5和MMCP-4的表达明显增加，Ang II的形成活性也明显增强。这些作用被过氧化氢酶的加入明显抑制，并被过氧化氢酶和超氧化物歧化酶的结合进一步抑制。单独与过氧化氢孵育可显著增加血管紧张素转换酶II的形成活性，而与过氧化氢酶共同处理则可完全抑制该活性。此外，在GATA抑制条件下，MMCP-5和MMCP-4的表达水平被显著抑制，同型半胱氨酸诱导的糜酶活性降低。同型半胱氨酸通过氧化应激的机制增加肥大细胞糜酶的表达和活性。我们的结果表明，氧化应激与局部Ang II形成系统之间存在生化联系。高血压研究(2010年)33,149154；DOI：10.1038/hr.2009.205；2009年12月4日在线发布
Reactive oxygen species (ROS) such as hydrogen peroxide (H2O2), O-2(center dot-) and OH center dot participate in the pathogenesis of ischemia/reperfusion injury, inflammation and atherosclerosis. Our previous studies have suggested that increased angiotensin II (Ang II)forming chymase may be involved in the development of atherosclerosis. However, the regulatory mechanism of chymase expression has not yet been clarified. In this study, we tested whether oxidative stress upregulates mouse mast cell proteinase chymase, mouse mast cell proteinase ( MMCP)-5 or MMCP-4. We also examined the expression and activity of these proteins after treatment. Cultured mouse mastocytoma cells (MMC) displaying chymase-dependent Ang II-forming activity were treated with H2O2 and several aminothiols with or without anti-oxidants. The levels of MMCP-5 and MMCP-4 expression were determined by quantitative RT-PCR; the level of chymase-dependent Ang II-forming activity was measured by high performance liquid chromatography using Ang I as a substrate. Treatment of MMC with homocysteine (0.1-3 mmol l(-1)) significantly increased MMCP-5 and MMCP-4 expression, as well as Ang II-forming activity. These effects were significantly inhibited by the addition of catalase and further suppressed by the combination of catalase and superoxide dismutase. Incubation with hydrogen peroxide alone caused a significant increase in Ang II-forming activity, which was completely suppressed by co-treatment with catalase. Furthermore, MMCP-5 and MMCP-4 expression levels were drastically suppressed and chymase induction by homocysteine was diminished under the GATA-inhibited condition. Homocysteine increased mast cell chymase expression and activity through the mechanism of oxidative stress. Our results suggest that there is a biochemical link between oxidative stress and the local Ang II-forming system. Hypertension Research (2010) 33, 149-154; doi: 10.1038/hr.2009.205; published online 4 December 2009