Resveratrol inhibits oxygen-glucose deprivation-induced MMP-3 expression and cell apoptosis in primary cortical cells via the NF-κB pathway
Resveratrol inhibits oxygen-glucose deprivation-induced MMP-3 expression and cell apoptosis in primary cortical cells via the NF-κB pathway
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DOI：
10.3892/mmr.2014.2239
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发表时间：
2014-08-01
影响因子：
3.4
通讯作者：
Fei, Zhou
中科院分区：
文献类型：
--
作者：
Huang, Tao;Gao, Dakuan;Fei, Zhou
白藜芦醇（Resveratrol，Res）或反式-3，4 '，5-三羟基二苯乙烯（trans-3，4'，5-trihydroxystilbene）已被证明在脑缺血中发挥神经保护作用。本研究的目的是调查是否Res具有神经保护作用的原代皮层神经元进行短暂的氧-葡萄糖剥夺（OGD）通过抑制基因编码的基质溶素-1，也被称为基质金属蛋白酶-3（MMP-3）的表达，并通过抑制细胞凋亡。原代皮质细胞暴露于OGD，然后再给氧以诱导短暂缺血。在存在或不存在核因子（NF）-κ B抑制剂吡咯烷二硫代氨基甲酸酯（PDTC; 10 μ M）或500 μ M一氧化氮（NO）供体NOC-18的情况下，在短暂缺血期间将Res（50 μ M）加入培养基中。使用四唑还原（MTT）测定法评估细胞活力。流式细胞仪检测细胞凋亡情况。Western blot和RT-PCR检测MMP-3表达，Western blot检测诱导型一氧化氮合酶（iNOS）、NF-κ B B、caspase-3、切割型caspase-3、B细胞淋巴瘤2（Bcl-2）和Bcl-2相关X蛋白（Box）表达。采用分光光度法检测NO。我们发现，细胞活力显着降低瞬时OGD，这种效果被逆转的Res治疗。此外，OGD还可诱导细胞凋亡、Bax表达和caspase-3活化，抑制Bcl-2表达，Res可逆转OGD的上述作用。RES处理通过抑制NF-κ B表达显著降低了由短暂OGD诱导的MMP-3水平。此外，Res抑制OGD诱导的iNOS表达和NO合成。MMP-3的表达诱导NO衰减Res治疗和部分恢复外源性NO使用NOC-18。综上所述，这些发现表明OGD通过典型的凋亡信号传导和通过调节MMP-3的表达来诱导细胞凋亡; Res可以通过NF-κ B-iNOS/NO途径逆转OGD诱导的MMP-3表达和细胞凋亡。因此，Res可能是一种很有前途的治疗中风相关神经元损伤的药物。
Resveratrol (Res) or trans-3,4',5-trihydroxystilbene, has been proven to exert neuroprotective effects in cerebral ischemia. The aim of the present study was to investigate whether Res has neuroprotective effects in primary cortical neurons subjected to transient oxygen-glucose deprivation (OGD) via inhibiting the expression of the gene encoding stromelysin-1, also known as matrix metalloproteinase-3 (MMP-3), and via inhibiting cell apoptosis. Primary cortical cells were exposed to OGD, followed by reoxygenation to induce transient ischemia. Res (50 mu M) was added into the culture medium during transient ischemia in the presence or absence of the nuclear factor (NF)-kappa B inhibitor pyrrolidine dithiocarbamate (PDTC; 10 mu M) or 500 mu M of the nitric oxide (NO) donor NOC-18. Cell viability was assessed using the tetrazolium reduction (MTT) assay. Cell apoptosis was evaluated by flow cytometry. MMP-3 expression was analyzed by western blot and reverse transcription-polymerase chain reaction (RT-PCR), while the levels of inducible NO synthase (iNOS), NF-kappa B, caspase-3, cleaved caspase-3, B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Box) were assayed by western blot. NO was detected using a spectrophotometric method. We found that the cellular viability was significantly reduced by transient OGD and that this effect was reversed by Res treatment. In addition, OGD was shown to induce cell apoptosis, the expression of Bax and the activation of caspase-3, and inhibit the expression of Bcl-2, and these effects were also reversed by Res treatment. Res treatment significantly reduced the level of MMP-3 that was induced by transient OGD, via inhibition of NF-kappa B expression. In addition, Res inhibited iNOS expression and NO synthesis that were induced by OGD. MMP-3 expression induced by NO was attenuated by Res treatment and was partially restored by exogenous NO using NOC-18. Taken together, these findings indicate that OGD induces apoptosis through canonical apoptosis signaling and by modulating the expression of MMP-3; Res can reverse the OGD-induced MMP-3 expression and cell apoptosis via the NF-kappa B-iNOS/NO pathway. Therefore, Res may be a promising agent for the treatment of neuronal injury associated with stroke.