Inhibition of hydrogen sulfide production by gene silencing attenuates inflammatory activity of LPS-activated RAW264.7 cells
Inhibition of hydrogen sulfide production by gene silencing attenuates inflammatory activity of LPS-activated RAW264.7 cells
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DOI：
10.1007/s00253-013-5080-x
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发表时间：
2013-09-01
影响因子：
5
通讯作者：
Bhatia, Madhav
中科院分区：
文献类型：
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作者：
Badiei, Alireza;Rivers-Auty, Jack;Bhatia, Madhav
硫化氢是一种炎症介质，由巨噬细胞中的胱硫醚γ-裂解酶（CSE）的活性产生。以前，CSE的药理学抑制已被报道具有相互矛盾的结果，这可能是由于缺乏特异性的药理学试剂。因此，本研究使用了一种非常特异的小干扰RNA（siRNA）方法来抑制体外环境中CSE的产生。我们发现脂多糖（LPS）激活巨噬细胞导致CSE mRNA和蛋白水平升高，以及促炎细胞因子和一氧化氮（NO）的产生增加。我们成功地使用siRNA来特异性地降低活化的巨噬细胞中CSE mRNA和蛋白的水平。此外，LPS激活的巨噬细胞中的促炎细胞因子的水平显着低于siRNA转染的细胞相比，在未转染的控制。然而，通过转染细胞的NO的产生水平较高，表明CSE活性对NO产生具有抑制作用。这些发现表明，CSE酶在巨噬细胞的活化中具有至关重要的作用，并且其活性对这些细胞的NO产生具有抑制作用。
Hydrogen sulfide is an inflammatory mediator and is produced by the activity of the enzyme cystathionine gamma-lyase (CSE) in macrophages. Previously, pharmacological inhibition of CSE has been reported to have conflicting results, and this may be due to the lack of specificity of the pharmacological agents. Therefore, this study used a very specific approach of small interfering RNA (siRNA) to inhibit the production of the CSE in an in vitro setting. We found that the activation of macrophages by lipopolysaccharide (LPS) resulted in higher levels of CSE mRNA and protein as well as the increased production of proinflammatory cytokines and nitric oxide (NO). We successfully used siRNA to specifically reduce the levels of CSE mRNA and protein in activated macrophages. Furthermore, the levels of proinflammatory cytokines in LPS-activated macrophages were significantly lower in siRNA-transfected cells compared to those in untransfected controls. However, the production levels of NO by the transfected cells were higher, suggesting that CSE activity has an inhibitory effect on NO production. These findings suggest that the CSE enzyme has a crucial role in the activation of macrophages, and its activity has an inhibitory effect on NO production by these cells.