Yan Junqiang, Sun Jiachun, Huang Lina, Fu Qizhi, Du Ganqin
Department of Neurology, The First Affiliated Hospital of He Nan University of Science and Technology, Henan, People's Republic of China.
J Neurosci Res. 2014 May;92(5):634-40. doi: 10.1002/jnr.23329. Epub 2014 Jan 31.
This study investigates the impact of simvastatin on neuroinflammation in experimental parkinsonian cell models. 6-Hydroxydopamine (6-OHDA)-treated pheochromocytoma-12 (PC12) cells were used to investigate the neuroprotective nature of simvastatin. After incubation with 6-OHDA, simvastatin, and/or N-methyl-D-aspartic acid receptor 1 (NMDAR1) siRNA for 24 hr, test kits were used to detect the levels of lactate dehydrogenase (LDH) and glutamate released from PC12 cells exposed to different culture media. The mRNA levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 were determined by RT-PCR, and the protein levels were analyzed by Western blot. NMDAR1 were also determined by RT-PCR and the protein levels were analyzed by Western blot. LDH and glutamate levels in 6-OHDA-incubated PC12 cells increased compared with those in the controls, and incubation with simvastatin inhibited this elevation. Silencing of NMDAR1 with siRNA inhibited the expression of LDH and glutamate to a degree similar to simvastatin. The expression levels of NMDAR1, TNF-α, IL-1β, and IL-6 were significantly upregulated after treatment with 6-OHDA. The 6-OHDA-stimulated mRNA and protein levels of the proinflammatory cytokines NMDAR1, TNF-α, IL-1β, and IL-6 were reduced by simvastatin. Silencing of NMDAR1 with siRNA decreased the NMDAR1, TNF-α, IL-1β, and IL-6 mRNA and protein expression levels in 6-OHDA-stimulated PC12 cells. Simvastatin could also inhibit the expression of NMDAR1 and cytokines to a degree similar to silencing of NMDAR1 with siRNA. Our results suggest that NMDAR1 modulation could explain the anti-inflammatory mechanisms of simvastatin in experimental parkinsonian cell models.
本研究调查了辛伐他汀对实验性帕金森病细胞模型中神经炎症的影响。使用经6-羟基多巴胺（6-OHDA）处理的嗜铬细胞瘤-12（PC12）细胞来研究辛伐他汀的神经保护特性。在用6-OHDA、辛伐他汀和/或N-甲基-D-天冬氨酸受体1（NMDAR1）小干扰RNA（siRNA）孵育24小时后，使用检测试剂盒检测暴露于不同培养基的PC12细胞释放的乳酸脱氢酶（LDH）和谷氨酸水平。通过逆转录聚合酶链反应（RT-PCR）测定肿瘤坏死因子（TNF）-α、白细胞介素（IL）-1β和IL-6的mRNA水平，并通过蛋白质印迹法分析蛋白质水平。还通过RT-PCR测定NMDAR1并通过蛋白质印迹法分析蛋白质水平。与对照组相比，经6-OHDA孵育的PC12细胞中的LDH和谷氨酸水平升高，而用辛伐他汀孵育可抑制这种升高。用siRNA沉默NMDAR1在一定程度上抑制了LDH和谷氨酸的表达，其程度与辛伐他汀相似。用6-OHDA处理后，NMDAR1、TNF-α、IL-1β和IL-6的表达水平显著上调。辛伐他汀降低了6-OHDA刺激的促炎细胞因子NMDAR1、TNF-α、IL-1β和IL-6的mRNA和蛋白质水平。用siRNA沉默NMDAR1降低了6-OHDA刺激的PC12细胞中NMDAR1、TNF-α、IL-1β和IL-6的mRNA和蛋白质表达水平。辛伐他汀还可以在一定程度上抑制NMDAR1和细胞因子的表达，其程度与用siRNA沉默NMDAR1相似。我们的结果表明，NMDAR1调节可以解释辛伐他汀在实验性帕金森病细胞模型中的抗炎机制。
