Nrf2 Inhibits Periodontal Ligament Stem Cell Apoptosis under Excessive Oxidative Stress.
Nrf2 Inhibits Periodontal Ligament Stem Cell Apoptosis under Excessive Oxidative Stress.
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Nrf2抑制过度氧化应激下牙周膜干细胞凋亡
DOI：
10.3390/ijms18051076
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发表时间：
2017-05-17
影响因子：
5.6
通讯作者：
Chen Y
中科院分区：
文献类型：
--
作者：
Liu Y;Yang H;Wen Y;Li B;Zhao Y;Xing J;Zhang M;Chen Y
本研究旨在分析牙周炎氧化微环境中Nrf 2介导的牙周膜干细胞抗凋亡的新机制。我们用H2 O2处理的PDLSC建立了氧化应激模型。我们采用实时荧光定量PCR、Western blotting、TUNEL染色、荧光分析和转移遗传学等方法来确定氧化应激和凋亡的程度以及核因子-红细胞2相关因子2（Nrf 2）的功能。结果表明，随着活性氧（ROS）和丙二醛（MDA）水平的升高，过氧化氢（H2 O2）处理的氧化应激效应明显。氧化分子在H2 O2暴露后发生改变，Nrf 2的信号传导被激活，其下游效应物血红素加氧酶-1（HO-1）、NAD（P）H：醌氧化还原酶1（NQO 1）和γ-谷氨酰半胱氨酸合成酶（γ-GCS）增加。此外，凋亡水平随着氧化应激而逐渐增加，除了Bcl-2的下调外，caspase-9，caspase-3，Bax和c-Fos水平也上调。然而，caspase-8的水平没有改变。抗氧化作用的增强并不能减轻细胞凋亡的发生。此外，Nrf 2过表达有效地提高了抗氧化水平并增加了细胞增殖。同时，过表达有效地抑制TUNEL染色，降低caspase-9，caspase-3，Bax和c-Fos的分子水平，但不影响caspase-8。相反，沉默Nrf 2水平的表达具有相反的效果。总的来说，Nrf 2通过其调节氧化应激和通过激活氧化酶抗内源性凋亡的作用使PDLSC活化。
The present study aimed to analyze novel mechanisms underlying Nrf2-mediated anti-apoptosis in periodontal ligament stem cells (PDLSCs) in the periodontitis oxidative microenvironment. We created an oxidative stress model with H2O2-treated PDLSCs. We used real-time PCR, Western blotting, TUNEL staining, fluorogenic assay and transfer genetics to confirm the degree of oxidative stress and apoptosis as well as the function of nuclear factor-erythroid 2-related factor 2 (Nrf2). We demonstrated that with upregulated levels of reactive oxygen species (ROS) and malondialdehyde (MDA), the effect of oxidative stress was obvious under H2O2 treatment. Oxidative molecules were altered after the H2O2 exposure, whereby the signaling of Nrf2 was activated with an increase in its downstream effectors, heme oxygenase-1 (HO-1), NAD(P)H:quinone oxidoreductase 1 (NQO1) and γ-glutamyl cysteine synthetase (γ-GCS). Additionally, the apoptosis levels gradually increased with oxidative stress by the upregulation of caspase-9, caspase-3, Bax and c-Fos levels in addition to the downregulation of Bcl-2. However, there was no alterations in levels of caspase-8. The enhanced antioxidant effect could not mitigate the occurrence of apoptosis. Furthermore, Nrf2 overexpression effectively improved the anti-oxidative levels and increased cell proliferation. At the same time, overexpression effectively restrained TUNEL staining and decreased the molecular levels of caspase-9, caspase-3, Bax and c-Fos, but not that of caspase-8. In contrast, silencing the expression of Nrf2 levels had the opposite effect. Collectively, Nrf2 alleviates PDLSCs via its effects on regulating oxidative stress and anti-intrinsic apoptosis by the activation of oxidative enzymes.