Cerebral ischemia/reperfusion injury (IRI) is one of major causes of ischemic organ damage. It is well established that inflammatory cytokines serve as regulatory factors in cerebral oxygen glucose deprivation/reoxygenation (OGD/R). However, the involving mechanism is not clear enough. OGD/R PC12 cells were used as a hypoxia/reoxygenation model. IL-32 expression and cell viability were detected by qRT-PCR and CCK-8 assay, respectively. Cell apoptosis were determined by flow cytometry and western blotting. Protein levels of inflammatory factors, and the activity of MPO, MDA and SOD were analyzed. Furthermore, western blot assay was carried out to assess protein levels of Nrf2, keap1, NQO-1, p-p65, p-IκBα, p65 and IκBα. The results revealed that IL-32 expression was significantly upregulated in PC12 cells induced by OGD/R. Nrf2, keap1 and NQO-1 level was reduced while phosphorylation level of p65 and IκBα was up-regulated in OGD/R-induced PC12 cells. Mechanism investigations found that IL-32 silence elevated the level of Nrf2, Keap1 and NQO-1, reduced p-p65 and p-IκBα level, and regulated the contents of TNF-a, IL-1β, IL-6 and MCP-1 in OGD/R PC12 cells. In addition, knockdown of IL-32 suppressed production of intracellular ROS, elevated SOD activity, reduced MPO and MDA content, and enhanced cell viability. Furthermore, cell apoptosis was induced in OGD/R PC12 cells with IL-32 silence. However, Nrf2 inhibitor reversed the effects of IL-32 knockdown on OGD/R PC12 cells. This research suggests that IL-32 silence may alleviate OGD/R and Nrf2 plays an important role in the protection by IL-32 silence on PC12 cells induced by OGD/R.

中文翻译：

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抑制IL-32可通过激活Nrf2 /NF-κB途径保护PC12细胞免受氧葡萄糖剥夺/复氧诱导的损伤。

脑缺血/再灌注损伤（IRI）是缺血性器官损害的主要原因之一。众所周知，炎性细胞因子是脑氧葡萄糖剥夺/复氧（OGD / R）的调节因子。但是，参与机制还不够清楚。OGD / R PC12细胞用作缺氧/复氧模型。通过qRT-PCR和CCK-8测定分别检测IL-32表达和细胞生存力。通过流式细胞仪和蛋白质印迹法测定细胞凋亡。分析了炎症因子的蛋白水平以及MPO，MDA和SOD的活性。此外，进行了蛋白质印迹分析以评估Nrf2，keap1，NQO-1，p-p65，p-IκBα，p65和IκBα的蛋白质水平。结果表明，由OGD / R诱导的PC12细胞中IL-32表达明显上调。Nrf2，在OGD / R诱导的PC12细胞中，keap1和NQO-1水平降低，而p65和IκBα的磷酸化水平上调。机制研究发现，IL-32沉默可提高Nrf2，Keap1和NQO-1的水平，降低p-p65和p-IκBα的水平，并调节TNF-a，IL-1β，IL-6和MCP-1的含量在OGD / R PC12细胞中。此外，IL-32的抑制可抑制细胞内ROS的产生，SOD活性升高，MPO和MDA含量降低以及细胞活力增强。此外，在具有IL-32沉默的OGD / R PC12细胞中诱导细胞凋亡。但是，Nrf2抑制剂逆转了IL-32敲低对OGD / R PC12细胞的作用。这项研究表明，IL-32沉默可能减轻OGD / R，Nrf2在IL-32沉默对OGD / R诱导的PC12细胞的保护中起着重要作用。