NADPH oxidase 4 regulates vascular inflammation in aging and atherosclerosis.
NADPH oxidase 4 regulates vascular inflammation in aging and atherosclerosis.
复制标题
DOI：
10.1016/j.yjmcc.2016.12.004
复制
发表时间：
2017-01
影响因子：
5
通讯作者：
Runge MS
中科院分区：
文献类型：
--
作者：
Lozhkin A;Vendrov AE;Pan H;Wickline SA;Madamanchi NR;Runge MS
我们最近报道，在衰老过程中，NADPH氧化酶4(NOX4)的表达和活性增加会导致细胞和线粒体氧化应激、血管炎症、功能障碍和动脉粥样硬化。本研究的目的是阐明这些作用的分子机制(S)，并确定NOX4对小鼠血管平滑肌细胞(VSMCs)促炎基因表达的调节作用。一种新的多肽介导的siRNA转染法被用来抑制NOX4的表达，而细胞毒性最小。使用蜂毒素衍生的多肽p5RHH，我们获得了显著更高的转染率(92%比脂质体的85%)和更低的毒性(在MTT法中p<；0.001与脂质体相比，在乳酸脱氢酶检测中p<；0.0001与脂联胺)。转化生长因子β-1显著上调VSMC中NOX4mRNA和蛋白的表达。P5RHH介导的NOX4siRNA显著抑制转化生长因子β-1诱导的NOX4mRNA(p<；0.01)和蛋白(p<；0.0001)水平的上调，并减少过氧化氢的产生(p<；0.0001)。促炎症基因CCL2、CCL5、IL6和VCAM1在老年和幼年野生型小鼠的VSMCs、APOE−/−小鼠动脉粥样硬化的动脉和人颈动脉中的表达显著上调，并与NOX4的表达相关。P5RHH介导的NOX4siRNA可显著抑制转化生长因子β-1诱导的小鼠血管平滑肌细胞中这些促炎基因的表达，其中对IL-6表达的抑制程度最高。P5RHH抑制转化生长因子β激活的蛋白1(TAK1，又称MAP3K7)，Jun和Rela，而不是NFKB2，下调转化生长因子β1诱导的VSMC中NOX4的表达。综上所述，这些数据表明，NOX4的表达和激活增加，可能是由于衰老过程中转化生长因子β1水平的增加，诱导了血管平滑肌细胞的促炎表型，从而加强了动脉粥样硬化。
We recently reported that increased NADPH oxidase 4 (NOX4) expression and activity during aging results in enhanced cellular and mitochondrial oxidative stress, vascular inflammation, dysfunction, and atherosclerosis. The goal of the present study was to elucidate the molecular mechanism(s) for these effects and determine the importance of NOX4 modulation of proinflammatory gene expression in mouse vascular smooth muscle cells (VSMCs). A novel peptide-mediated siRNA transfection approach was used to inhibit Nox4 expression with minimal cellular toxicity. Using melittin-derived peptide p5RHH, we achieved significantly higher transfection efficiency (92% vs. 85% with Lipofectamine) and decreased toxicity (p < 0.001 vs. Lipofectamine in MTT and p < 0.0001 vs. Lipofectamine in LDH assays) in VSMCs. TGFβ1 significantly upregulates Nox4 mRNA (p < 0.01) and protein (p < 0.01) expression in VSMCs. p5RHH-mediated Nox4 siRNA transfection greatly attenuated TGFβ1-induced upregulation of Nox4 mRNA (p < 0.01) and protein (p < 0.0001) levels and decreased hydrogen peroxide production (p < 0.0001). Expression of pro-inflammatory genes Ccl2, Ccl5, Il6, and Vcam1 was significantly upregulated in VSMCs in several settings cells isolated from aged vs. young wild-type mice, in atherosclerotic arteries of Apoe−/− mice, and atherosclerotic human carotid arteries and correlated with NOX4 expression. p5RHH-mediated Nox4 siRNA transfection significantly attenuated the expression of these pro-inflammatory genes in TGFβ1-treated mouse VSMCs, with the highest degree of inhibition in the expression of Il6. p5RHH peptide-mediated knockdown of TGFβ-activated kinase 1 (TAK1, also known as Map3k7), Jun, and Rela, but not Nfkb2, downregulated TGFβ1-induced Nox4 expression in VSMCs. Together, these data demonstrate that increased expression and activation of NOX4, which might result from increased TGFβ1 levels seen during aging, induces a proinflammatory phenotype in VSMCs, enhancing atherosclerosis.
登录
查看更多内容
TGF-β和活性氧的相互调节：纤维化的不良周期。
影响因子：
11.4
作者：
Liu RM;Desai LP
通讯作者：
Desai LP
影响因子：
20.1
作者：
Libby P;Hansson GK
通讯作者：
Hansson GK
影响因子：
14.8
作者：
通讯作者：
--
影响因子：
7.4
作者：
Craige SM;Kant S;Reif M;Chen K;Pei Y;Angoff R;Sugamura K;Fitzgibbons T;Keaney JF Jr
通讯作者：
Keaney JF Jr
影响因子：
3.5
作者：
Bai G;Hock TD;Logsdon N;Zhou Y;Thannickal VJ
通讯作者：
Thannickal VJ