以神經微膠細胞發炎反應為標的之阿茲海默氏症治療策略
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2017
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阿茲海默氏症是常見的漸進性神經退化性疾病，主要病理特徵包括細胞外間質的β類澱粉樣纖維(Aβ fibril)堆積，及細胞內高度磷酸化tau形成的神經纖維糾結。目前對於阿茲海默氏症致病機制仍未完全了解，患者也無法被完全治癒，因此發展阿茲海默氏症的治療策略是相當重要的。近年研究顯示神經發炎反應在阿茲海默氏症上扮演重要的角色，因此抑制神經發炎反應已成為治療阿茲海默氏症的重要標的。與發炎反應相關的微膠細胞是中樞神經的免疫細胞，具有典型的M1 (發炎反應)及替代的M2 (免疫抑制)兩種活化路徑，因此調節微膠細胞活化狀態已被認為是治療神經發炎性疾病的策略。先前本實驗室於脂多醣(LPS)刺激的RAW264.7小鼠巨噬細胞實驗，發現化合物VB-030、037能降低一氧化氮(NO)、TNF-α、IL-1β、PEG2等發炎因子的釋放。延續此發現，本研究首先利用LPS或β類澱粉樣纖維刺激BV-2微膠細胞模式，探究化合物VB-030、037的抗發炎能力，結果發現VB-037能顯著降低LPS/β類澱粉樣纖維刺激BV-2細胞之NO及Iba1蛋白表現，表示VB-037具有抗β類澱粉樣纖維誘導細胞發炎的潛力，同時也利用小鼠發炎抗體矩陣(Mouse inflammation antibody array)揭示VB-037之抗發炎標的IL-1α，並利用西方轉漬分析證實VB-037能顯著降低活化之BV-2細胞之促發炎因子IL-1α。為了測試VB-037對於神經細胞的保護效果，我們利用受發炎刺激之BV-2細胞制約培養液或共處理發炎因子LPS與IFN-γ，建構誘導表現Aβ-GFP融合蛋白的人類SH-SY5Y神經母細胞瘤細胞發炎模式，實驗結果發現VB-037能提升表現Aβ-GFP融合蛋白的SH-SY5Y之細胞存活率，降低細胞損傷後之乳酸脫氫酶(LDH)釋放量，並促進神經突生長，同時前處理VB-037能降低受LPS與IFN-γ刺激之SH-SY5Y細胞IL1A及磷酸化P38 (T180/Y182)、JNK (T183/Y185)、JUN (S63、S73)蛋白表現量。綜合本研究的發現，VB-037具有抗β類澱粉樣纖維誘導之神經發炎作用及神經保護作用，有潛能發展為抗β類澱粉樣纖維誘導神經發炎之新穎藥物。
Alzheimer’s disease (AD) is a progressive neurodegenerative disease characterized by the presence of neurofibrillary tangles and amyloid plaques. Studies have shown that activated microglia are parallel to the disease region where pro-inflammatory cytokines are highly expressed, indicating that neuroinflammation may play an important role in AD. Microglia are the resident immune cell in the central nervous system and possess states of M1 (pro-inflammatory response) and M2 (immunosuppressive response). Regulating the activation state of microglia has been considered a potential therapy for neuroinflammatory disorders. Previously our laboratory found that compounds VB-030 and 037 reduced lipopolysaccharide (LPS)-induced release of NO, TNF-α, IL-1β and PEG2 from mouse macrophage RAW264.7 cells. In this study, anti-inflammation abilities of these two compounds were firstly tested by using BV-2 mouse microglia. VB-037 exhibited anti-inflammation activity by reducing NO release and Iba1 expression in LPS or Aβ fibril-stimulated BV-2 microglia. The therapeutic target of VB-037, IL-1α, was identified in BV-2 microglia using mouse inflammation antibody array. VB-037 further protected SH-SY5Y cells expressing Aβ-GFP from cell death and reduced lactate dehydrogenase (LDH) release induced by LPS/IFN-γ-stimulated BV-2 conditioned media or LPS and IFN-γ co-stimulation. VB-037 also promoted neurite outgrowth in Aβ-GFP expressing SH-SY5Y cells. The reduced expressions of IL1A, phospho-P38 (T180/Y182), phospho-JNK (T183/Y185), phospho-JUN (S63, S73) by VB-037 pre-treatment were further confirmed by Western blotting. These results indicated that VB-037 possessed anti-inflammation and neuro-protection effect on inflamed Aβ-GFP SH-SY5Y cells. In conclusion, VB-037 could be a novel therapeutic molecule on Alzheimer’s disease based on the anti-inflammation and neuro-protection effects.
Alzheimer’s disease (AD) is a progressive neurodegenerative disease characterized by the presence of neurofibrillary tangles and amyloid plaques. Studies have shown that activated microglia are parallel to the disease region where pro-inflammatory cytokines are highly expressed, indicating that neuroinflammation may play an important role in AD. Microglia are the resident immune cell in the central nervous system and possess states of M1 (pro-inflammatory response) and M2 (immunosuppressive response). Regulating the activation state of microglia has been considered a potential therapy for neuroinflammatory disorders. Previously our laboratory found that compounds VB-030 and 037 reduced lipopolysaccharide (LPS)-induced release of NO, TNF-α, IL-1β and PEG2 from mouse macrophage RAW264.7 cells. In this study, anti-inflammation abilities of these two compounds were firstly tested by using BV-2 mouse microglia. VB-037 exhibited anti-inflammation activity by reducing NO release and Iba1 expression in LPS or Aβ fibril-stimulated BV-2 microglia. The therapeutic target of VB-037, IL-1α, was identified in BV-2 microglia using mouse inflammation antibody array. VB-037 further protected SH-SY5Y cells expressing Aβ-GFP from cell death and reduced lactate dehydrogenase (LDH) release induced by LPS/IFN-γ-stimulated BV-2 conditioned media or LPS and IFN-γ co-stimulation. VB-037 also promoted neurite outgrowth in Aβ-GFP expressing SH-SY5Y cells. The reduced expressions of IL1A, phospho-P38 (T180/Y182), phospho-JNK (T183/Y185), phospho-JUN (S63, S73) by VB-037 pre-treatment were further confirmed by Western blotting. These results indicated that VB-037 possessed anti-inflammation and neuro-protection effect on inflamed Aβ-GFP SH-SY5Y cells. In conclusion, VB-037 could be a novel therapeutic molecule on Alzheimer’s disease based on the anti-inflammation and neuro-protection effects.
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阿茲海默氏症, 微膠細胞, 發炎反應, 喹啉衍生物, Alzheimer’s disease, microglia, inflammation, quinoline derivatives