BACKGROUND Idebenone is a well-appreciated mitochondrial protectant while the mechanisms underlying the neuroprotection in cerebral ischemia and reperfusion (I/R) remain elusive. It has been manifested NLRP3 inflammasom activation contributed to I/R induced damage. It raises questions how exactly NLRP3 inflammasom was activated in microglia and neuron and whether idebenone reverses the process in I/R. METHODS I/R rat model was utilized and BV2, primary microglia and PC12 cells were subjected to oxygen-glucose deprivation (OGD). Then, western-blotting, q-PCR, immunofluorescence staining, ELISA, flow cytometry and immunoprecipitation analysis were performed. RESULTS We found ROS-NLRP3 singaling was activated in BV2 cells at OGD/R 24 h. Importantly, microglial NLRP3 activation was essential for NLRP3 activation in PC12 cells under microglial-neuronal co-culture circumstance, which has been confirmed to induced neuronal apoptosis. Further, we found mitochondrial dysfunction in OGD/R led to mt-DNA translocation as well as generation of mt-ROS, resulting cytosolic accumulation of oxidized mt-DNA. Ultimately, oxidized mt-DNA binding to NLRP3 contributed to further activation of NLRP3 and dramatically augmented inflammation in BV2 and PC12 cells. Furthermore, idebenone treatment inhibited the process, thus suppressing the NLRP3-mediated inflammatory injury after OGD/R. In vivo, NLRP3 was activated in microglia of I/R rats and inhibition of NLRP3 was observed in idebenone treatment group, which had less neurological deficit and less infarct volume. INTERPRETATION Our data revealed the anti-inflammatory effects of idebenone via suppressing activation of NLRP3 and ameliorating NLRP3-mediating damage in I/R, which may provide new insight in therapeutic strategy for ischemic stroke.

中文翻译：

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艾地苯醌可通过抑制NLRP3炎症小体活性来减轻缺血和再灌注中的脑炎性损伤。

背景技术艾地苯醌是一种广为人知的线粒体保护剂，而在脑缺血和再灌注（I / R）中神经保护的潜在机制仍然难以捉摸。已经表明NLRP3炎性激素活化导致了I / R诱导的损伤。这就引发了一个问题，即小胶质细胞和神经元中NLRP3炎性激素的激活方式如何以及艾地苯醌是否会逆转I / R过程。方法采用I / R大鼠模型，对BV2，原发性小胶质细胞和PC12细胞进行氧葡萄糖剥夺（OGD）。然后，进行蛋白质印迹，q-PCR，免疫荧光染色，ELISA，流式细胞术和免疫沉淀分析。结果我们发现在OGD / R 24 h BV2细胞中激活了ROS-NLRP3信号。重要的，在神经胶质-神经元共培养的情况下，小胶质细胞的NLRP3激活对于PC12细胞中NLRP3的激活是必不可少的，这已被证实可诱导神经元凋亡。此外，我们发现OGD / R中的线粒体功能障碍导致mt-DNA易位以及mt-ROS的产生，从而导致氧化的mt-DNA发生胞质积累。最终，氧化的mt-DNA与NLRP3的结合有助于NLRP3的进一步活化，并显着增加BV2和PC12细胞的炎症。此外，艾地苯醌处理抑制了该过程，从而抑制了OGD / R后NLRP3介导的炎症损伤。在体内，NLRP3在I / R大鼠的小胶质细胞中被激活，在艾地苯醌治疗组中观察到NLRP3的抑制，这具有较少的神经系统缺陷和较小的梗塞体积。