Chen Jingkao, Yin Wei, Tu Yalin, Wang Shengnan, Yang Xiaohong, Chen Qiuhe, Zhang Xiao, Han Yifan, Pi Rongbiao
School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou 510006, China; International Joint Laboratory (SYSU-PolyU HK) of Novel Anti-dementia Drugs of Guangdong, Guangzhou 510006, China; Guangdong Province Key Laboratory of Brain Function and Disease, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, China.
Department of Biochemistry, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou 510080, China.
Eur J Pharmacol. 2017 Jul 5;806:1-9. doi: 10.1016/j.ejphar.2017.03.025. Epub 2017 Mar 16.
Microglia and astrocytes are largely responsible for inflammatory injury in the brain of Alzheimer's disease (AD). Increasing evidence has indicated that Rho kinase (ROCK) plays an important role in the regulation of neuroinflammation. Previously, we synthesized a new chemical entity L-F001 and proved its potential inhibitory effects on ROCK and oxidative stress. Here, we investigated the anti-inflammatory effects and the molecular mechanisms of L-F001 in vitro and in vivo. L-F001 remarkably suppressed lipopolysaccharides (LPS)-elevated expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) as well as LPS-induced production of nitric oxide (NO), reactive oxygen species, interleukin-6 (IL-6) and tumor necreactive oxygen speciesis factor-α (TNF-α) in microglial BV-2 cells and in cultured astrocytes. Furthermore, L-F001 inhibited the degradation of IκB and nuclear translocation of nuclear factor kappa B (NF-κB) p65 subunit. Moreover, L-F001 induced the upregulation of heme-oxygenase-1 (HO-1) and glutamate cysteine ligase modifier subunit (GCLM) expression, two downstream effectors of nuclear factor (erythroid-derived 2)-like 2 (Nrf2). It was interesting that L-F001 also activated phosphatidylinositol 3-kinase (PI3K) pathway and induced M1 (CD16/32, M1 marker)/ M2 (CD206, M2 maker) transition in BV-2 cells which was significantly blocked by a PI3K inhibitor, wortmannin. Finally, L-F001 markedly attenuated the level of pro-inflammatory mediators in a murine model of systemic acute brain inflammation induced by LPS. Taken together, these results indicate that the novel multifunctional ROCK inhibitor L-F001 suppresses neuroinflammation in vitro and in vivo via NF-κB inhibition and Nrf2 activation, suggesting that L-F001 may be a promising drug candidate for treating neuroinflammation-associated CNS diseases, including AD.
小胶质细胞和星形胶质细胞在阿尔茨海默病（AD）大脑的炎症损伤中起主要作用。越来越多的证据表明，Rho激酶（ROCK）在神经炎症调节中起重要作用。此前，我们合成了一种新的化学实体L-F001，并证明了其对ROCK和氧化应激的潜在抑制作用。在此，我们研究了L-F001在体外和体内的抗炎作用及分子机制。L-F001显著抑制脂多糖（LPS）升高的诱导型一氧化氮合酶（iNOS）和环氧化酶-2（COX-2）的表达以及LPS诱导的小胶质细胞BV-2细胞和培养的星形胶质细胞中一氧化氮（NO）、活性氧、白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）的产生。此外，L-F001抑制IκB的降解和核因子κB（NF-κB）p65亚基的核转位。此外，L-F001诱导血红素加氧酶-1（HO-1）和谷氨酸半胱氨酸连接酶修饰亚基（GCLM）表达上调，这是核因子（红系衍生2）样2（Nrf2）的两个下游效应分子。有趣的是，L-F001还激活磷脂酰肌醇3-激酶（PI3K）途径并诱导BV-2细胞中M1（CD16/32，M1标志物）/M2（CD206，M2标志物）转变，而PI3K抑制剂渥曼青霉素可显著阻断这种转变。最后，L-F001显著降低了LPS诱导的全身性急性脑炎症小鼠模型中促炎介质的水平。综上所述，这些结果表明新型多功能ROCK抑制剂L-F001通过抑制NF-κB和激活Nrf2在体外和体内抑制神经炎症，提示L-F001可能是治疗包括AD在内的神经炎症相关中枢神经系统疾病的有前景的候选药物。
