Song Gyun Jee, Nam Youngpyo, Jo Myungjin, Jung Myungsu, Koo Ja Young, Cho Wansang, Koh Minseob, Park Seung Bum, Suk Kyoungho
Department of Pharmacology, Brain Science & Engineering Institute, BK21 Plus KNU Biomedical Convergence Program, Kyungpook National University School of Medicine, Daegu, Republic of Korea.
Department of Chemistry, Seoul National University, Seoul, Republic of Korea.
Neuropharmacology. 2016 Oct;109:159-169. doi: 10.1016/j.neuropharm.2016.06.009. Epub 2016 Jun 8.
Neuroinflammation is a key process for many neurodegenerative diseases. Activated microglia and astrocytes play an essential role in neuroinflammation by producing nitric oxide (NO), inflammatory cytokines, chemokines, and neurotoxins. Therefore, targeting glia-mediated neuroinflammation using small-molecules is a potential therapeutic strategy. In this study, we performed a phenotypic screen using microglia cell-based assay to identify a hit compound containing N-carbamoylated urethane moiety (SNU-BP), which inhibits lipopolysaccharide (LPS)-induced NO production in microglia. SNU-BP inhibited pro-inflammatory cytokines and inducible nitric oxide synthase in LPS-stimulated microglia, and potentiated interleukin-4-induced arginase-1 expression. PPAR-γ was identified as a molecular target of SNU-BP. The PPAR response element reporter assay revealed that SNU-BP specifically activated PPAR-γ, but not PPAR-δ or -α, confirming that PPAR-γ is the target protein of SNU-BP. The anti-inflammatory effect of SNU-BP was attenuated by genetic and pharmacological inhibition of PPAR-γ. In addition, SNU-BP induced an anti-inflammatory phenotype in astrocytes as well, by inhibiting pro-inflammatory NO and TNF-α, while increasing anti-inflammatory genes, such as arginase-1 and Ym-1. Finally, SNU-BP exhibited an anti-inflammatory effect in the LPS-injected mouse brain, demonstrating a protective potential for neuroinflammatory diseases.
神经炎症是许多神经退行性疾病的关键过程。活化的小胶质细胞和星形胶质细胞通过产生一氧化氮（NO）、炎性细胞因子、趋化因子和神经毒素在神经炎症中发挥重要作用。因此，使用小分子靶向胶质细胞介导的神经炎症是一种潜在的治疗策略。在本研究中，我们使用基于小胶质细胞的检测方法进行了表型筛选，以鉴定一种含有N-氨基甲酰化氨基甲酸酯部分的命中化合物（SNU-BP），其可抑制小胶质细胞中脂多糖（LPS）诱导的NO产生。SNU-BP抑制LPS刺激的小胶质细胞中的促炎细胞因子和诱导型一氧化氮合酶，并增强白细胞介素-4诱导的精氨酸酶-1表达。PPAR-γ被鉴定为SNU-BP的分子靶点。PPAR反应元件报告基因检测显示，SNU-BP特异性激活PPAR-γ，而不激活PPAR-δ或-α，证实PPAR-γ是SNU-BP的靶蛋白。SNU-BP的抗炎作用通过PPAR-γ的基因和药理学抑制而减弱。此外，SNU-BP还通过抑制促炎NO和TNF-α，同时增加抗炎基因（如精氨酸酶-1和Ym-1），在星形胶质细胞中诱导抗炎表型。最后，SNU-BP在注射LPS的小鼠脑中表现出抗炎作用，显示出对神经炎症性疾病的保护潜力。
