榛仁球蛋白對脂多醣所誘導小鼠腦部微小膠細胞活化之抗發炎作用

研究背景:發炎反應與許多疾病的發生有關聯性，其中退化性神經疾病的病理進程已被證實與發炎反應有關，而腦中微小膠細胞是造成發炎反應最主要的細胞，許多研究指出，抑制腦中的發炎反應能有效減緩退化性神經疾病的病理進程。因此，本研究使用小鼠腦中的微小膠細胞進行實驗，利用革蘭氏陰性菌的脂多醣（Lipopolysaccharide；LPS）刺激微小膠細胞產生發炎反應，以模擬神經退化性疾病的病理模式，如阿茲海默氏症與巴金森氏症等疾病。研究目的：探討補骨脂所萃取出來的天然化合物榛仁球蛋白（Corylin）其抑制LPS誘導微小膠細胞發炎反應的成效及作用機轉。研究方法：測試Corylin對抑制LPS誘發腦中微小膠細胞產生一氧化氮的能力，並找出有效的作用濃度。此外，為避免Corylin使用的劑量對細胞有毒性，我們也測試Corylin對細胞存活之影響。接著使用酵素結合免疫吸附分析法來探討Corylin對抑制LPS誘發腦中微小膠細胞產生發炎相關細胞激素（TNF-α、IL-1β與IL-6）的作用。最後本研究使用西方墨點法分析發炎相關的分子機制，進一步探討Corylin對抑制LPS誘發腦中微小膠細胞產生發炎反應的作用機轉。研究結果：本研究結果顯示，Corylin能有效的抑制LPS所誘發的發炎反應，其中Corylin透過抑制LPS誘導MAPKs (JNK，p38 MAPK和ERK)的磷酸化，進而使iNOS與COX-2表現量降低，並且抑制促發炎細胞激素(TNF-α、IL-6與IL-1β)的釋放。此外，Corylin能藉由抑制LPS誘導NLRP3 inflammasome的活性，使微小膠細胞內NLRP3、ASC、Caspase-1與IL-1β的蛋白質表現下降。結論：實驗結果顯示Corylin會抑制LPS誘導的腦部炎症反應，具有改善發炎相關神經退化性疾病的治療潛力。

關鍵字

榛仁球蛋白；微小膠細胞；抗發炎

並列摘要

Background：Inflammation has been found to be associated with many neurodegenerative diseases, including parkinsonism and dementia. In the present study, we used lipopolysaccharide (LPS) to simulate murine microglia cells (BV2 cells) as an experimental model to mimic the inflammatory environment in brain. Aim: We used LPS-activated BV2 cells as an experimental model to examine the anti-inflammatory ability of corylin, a main compound isolated from Psoralea corylifolia L. which commonly used in Chinese herbal medicine. Method：The production of NO by LPS-activated BV2 cells was measured using Greiss reaction. The secretion of proinflammatory cytokines including tumor necrosis factor (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) by LPS-activated BV2 cells was analyzed using ELISA. The expression of iNOS, COX-2, NLRP3, IL-1β and MAPKs in LPS-activated BV2 cells was examined by Western blot. Result：Our experimental results showed that corylin suppressed the production of proinflammatory cytokines by LPS-activated BV2 cells. In addition, corylin inhibited the expression levels of iNOS and COX-2, attenuated the phosphorylation of ERK, JNK and p38, inhibited the activation of caspase-1 and IL-1β, and decreased the expression levels of NLRP3 (Nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing-3 OR Nod-like receptor protein 3) and ASC (Apoptosis-associated Speck-like protein containing a caspase activation and recruitment domain) by LPS-activated BV2 cells. These results indicate the anti-inflammatory effects of corylin may act through attenuating LPS-induced inflammation and inhibiting the activation of NLRP3 inflammasome in LPS-activated BV2 cells. Conclusion：Our experimental results suggest that corylin might have potential in treating brain inflammation and attenuating the progression of neurodegeneration diseases.

並列關鍵字

Corylin ； Microglia cells ； Anti-inflammatory

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榛仁球蛋白對脂多醣所誘導小鼠腦部微小膠細胞活化之抗發炎作用

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