Zhou Qin, Guo Dongkai, Li Xinjia, Wang Yixuan, Ye Xiaoli, Xue Sudong, Wang Xiaoyu
Department of Pharmacy, The Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou Municipal Hospital, Suzhou, Jiangsu, 215002, P.R. China.
Department of pharmacy, The Affiliated Suzhou Science and Technology Town Hospital of Nanjing Medical University, Suzhou, Jiangsu, 215153, P.R. China.
An Acad Bras Cienc. 2020 Nov 20;92(4):e20200241. doi: 10.1590/0001-3765202020200241. eCollection 2020.
Microglia are the resident immune cells in the central nervous system (CNS), which play important roles in the repair of neuroinflammatory injury. The present study investigated the anti-neuroinflammatory effects of vinpocetine induced by lipopolysaccharide (LPS) in BV2 microglia. BV2 microglia were pretreated with vinpocetine, and then stimulated with LPS (100 ng/mL). The cytotoxicity of BV2 microglia was assessed by MTT assay. The expression levels of nitrite oxide were measured by Griess assay. Proinflammatory cytokines and mediators were determined by Western blot, ELISA, or quantitative real-time PCR. Vinpocetine significantly decreased the generation of nitric oxide-inducible nitric oxide synthase (iNOS), cyclooxygenase- (COX-) 2 in a dose-dependent manner. In addition, vinpocetine decreased the production of pro-inflammatory cytokines such as tumor necrosis factor alpha (TNF-α), interleukin (IL)-6 and IL-1β. Furthermore, it was observed that phosphorylation levels of AMPK (Thr-172) decreased in LPS-stimulated BV2 microglia. Vinpocetine treatment increased AMPK phosphorylation in LPS-stimulated BV2 microglia. AMPK inhibition by siRNA blocked the anti-inflammatory effects of vinpocetine induced by LPS in BV2 microglia. The overall results demonstrate that vinpocetine has anti-inflammatory effects on LPS-stimulated BV2 microglia via inducing phosphorylation of AMPK, suggesting that vinpocetine is a potential therapeutic agent in neuroinflammatory injury.
小胶质细胞是中枢神经系统（CNS）中的常驻免疫细胞，在神经炎性损伤修复中发挥重要作用。本研究调查了长春西汀对脂多糖（LPS）诱导的BV2小胶质细胞的抗神经炎症作用。用长春西汀预处理BV2小胶质细胞，然后用LPS（100 ng/mL）刺激。通过MTT法评估BV2小胶质细胞的细胞毒性。通过格里斯试剂法测量亚硝酸盐的表达水平。通过蛋白质免疫印迹法、酶联免疫吸附测定法或定量实时聚合酶链反应来测定促炎细胞因子和介质。长春西汀以剂量依赖性方式显著降低一氧化氮诱导型一氧化氮合酶（iNOS）、环氧化酶-（COX-）2的生成。此外，长春西汀降低了促炎细胞因子如肿瘤坏死因子α（TNF-α）、白细胞介素（IL）-6和IL-1β的产生。此外，观察到在LPS刺激的BV2小胶质细胞中，AMPK（Thr-172）的磷酸化水平降低。长春西汀处理增加了LPS刺激的BV2小胶质细胞中AMPK的磷酸化。通过小干扰RNA抑制AMPK可阻断长春西汀对LPS诱导的BV2小胶质细胞的抗炎作用。总体结果表明，长春西汀通过诱导AMPK磷酸化对LPS刺激的BV2小胶质细胞具有抗炎作用，提示长春西汀是神经炎性损伤的一种潜在治疗药物。
