Pien-Tze-Huang, a Chinese patent formula, attenuates NLRP3 inflammasome-related neuroinflammation by enhancing autophagy via the AMPK/mTOR/ULK1 signaling pathway
Pien-Tze-Huang, a Chinese patent formula, attenuates NLRP3 inflammasome-related neuroinflammation by enhancing autophagy via the AMPK/mTOR/ULK1 signaling pathway
复制标题
中成药片子黄通过AMPK/mTOR/ULK1信号通路增强自噬，从而减轻NLRP3炎症相关的神经炎症
DOI：
10.1016/j.biopha.2021.111814
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发表时间：
2021-06-17
影响因子：
7.5
通讯作者：
Huang, Mingqing
中科院分区：
文献类型：
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作者：
Huang, Zhenwei;Zhou, Xian;Huang, Mingqing
NLRP 3炎性体是缺血性卒中诱导的神经炎症和随后的脑损伤的关键介质。我们前期的研究证实了著名中成药片仔癀（PTH）对脑缺血再灌注损伤大鼠海马神经元凋亡的抑制作用。本研究旨在阐明甲状旁腺素（PTH）在脂多糖（LPS）诱导的BV 2小胶质细胞和脑缺血/再灌注损伤大鼠神经炎症中的作用机制。用LPS刺激BV 2细胞12小时，并用不同浓度的PTH处理。分别通过实时PCR和蛋白质印迹分析PTH对相关基因（IL-6、IL-1 β、IL-18、TNF-α、考克斯-2和iNOS mRNA）和蛋白质（NLRP 3炎性体、自噬和AMPK/mTOR/ULK信号传导）的调节。在大脑中动脉闭塞大鼠模型中进行了类似的分析，包括神经功能缺损、梗死体积、小胶质细胞活化以及调节自噬和NLRP 3的关键基因和蛋白。我们的研究结果表明，PTH显着抑制LPS诱导的BV 2细胞中关键的促炎介质的产生和NLRP 3和caspase-1 p20的蛋白表达。它还通过AMPK/mTOR/ULK相关通路调节关键的自噬蛋白来增强自噬反应。自噬抑制剂（3-MA）和AMPK阻断剂（化合物C）部分阻断了PTH引起的炎症反应和NLRP 3表达的降低。在大鼠中，甲状旁腺素显著减少梗死面积，抑制小胶质细胞活化，改善神经元缺损。它还促进自噬并降低NLRP 3活性。我们的研究表明，PTH抑制NLRP 3炎性小体介导的神经炎症，这是通过AMPK/mTOR/ULK 1途径增强自噬在体外和体内。
NLRP3 inflammasome is a key mediator in ischemic stroke-induced neuroinflammation and subsequent brain injury. Our previous study demonstrated the potent activity of Pien-Tze-Huang (PTH), a well-known Chinese patent formula, in reducing mitochondria-mediated neuronal apoptosis in cerebral ischemia/reperfusion impaired rats. This study aims to elucidate the mechanistic action of PTH related to neuroinflammation in LPSinduced BV2 microglial cells and cerebral ischemia/reperfusion impaired rats. BV2 cells were stimulated with LPS for 12 h and treated with PTH with various concentrations. Modulation by PTH of relevant genes (IL-6, IL-1 beta, IL-18, TNF-alpha, COX-2 and iNOS mRNA) and proteins (NLRP3 inflammasome, autophagy and AMPK/mTOR/ULK signaling) was analyzed by real-time PCR and western blot, respectively. Similar analyses were conducted in middle cerebral artery occlusion rat model including neurological deficit, infarct volume, microglial activation, and key genes and proteins in modulating autophagy and NLRP3. Our results showed that PTH significantly inhibited the production of key proinflammatory mediators and protein expressions of NLRP3 and caspase-1 p20 in LPS induced BV2 cells. It also enhanced the autophagy response by modulating the key autophagy proteins via AMPK/mTOR/ULK related pathway. The reduced inflammatory responses and NLRP3 expressions by PTH were partially blocked by the autophagy inhibitor (3-MA) and AMPK blocker (compound C). In rats, PTH significantly reduced infarct size, suppressed microglial activation, and improved neuron deficit. It also promoted autophagy and reduced NLRP3 activity. Our study demonstrated that PTH inhibited NLRP3 inflammasome-mediated neuroinflammation, which was associated with enhanced autophagy via AMPK/mTOR/ULK1 pathway in vitro and in vivo.