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標題: n-butylidenephthalide減少唐氏症誘導萬能幹細胞分化之神經細胞的類阿茲海默症病徵
n-butylidenephthalide Attenuates Alzheimer's Disease-Like Cytopathy in Down Syndrome Induced Pluripotent Stem Cell-Derived Neurons
作者: 張嘉佑
Chia-Yu Chang
關鍵字: induced pluripotent stem cells;neural stem cells;誘導萬能幹細胞;神經幹細胞;阿茲海默症
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Alzheimer's disease (AD) is a critical neuronal degenerative disease that still has no functional cure yet. Down syndrome (DS) patients with early-onset dementia share similar neurodegenerative features with Alzheimer's disease (AD). To recapitulate the AD cell model for drug screening, we first developed an economical and efficiency culture method for long term culture of human pluripotent stem cells. DS induced pluripotent stem cells (DS-iPSCs), reprogrammed from mesenchymal stem cells in amniotic fluid, were maintained in our culture system and directed toward a neuronal lineage. Neuroepithelial precursor cells with high purity and forebrain characteristics were robustly generated on day 10 (D10) of differentiation. Accumulated amyloid deposits, Tau protein hyperphosphorylation and Tau intracellular redistribution emerged rapidly in DS neurons within 45 days but not in normal embryonic stem cell-derived neurons. n-butylidenephthalide (Bdph), a major phthalide ingredient of Angelica sinensis, was emulsified by pluronic F127 to reduce its cellular toxicity and promote canonical Wnt signaling. Interestingly, we found that F127-Bdph showed significant therapeutic effects in reducing secreted Aβ40 deposits, the total Tau level and the hyperphosphorylated status of Tau in DS neurons. Taken together, our culture system can keep DS-iPSC with highly pluripotency and the DS-iPSC derived neural cells can serve as an ideal cellular model of DS and AD and have potential for high-throughput screening of candidate drugs. We also suggest that Bdph may benefit DS or AD treatment by scavenging Aβ aggregates and neurofibrillary tangles.

阿茲海默症為一種嚴重的神經退化性疾病,並且目前尚無有效的醫療方式。唐氏症患者常會罹患早發性的失智症並和阿茲海默症有類似的病理特徵。為了建立高效率的阿茲海默症細胞模型並進行藥物篩選,我們首先發展了一種較為便宜且有效率的人類萬能幹細胞無滋養層培養系統。由唐氏症胎兒羊水中取得的間質幹細胞經由重新編程後,回復成為誘導性萬能幹細胞。我們將此唐氏症誘導性萬能幹細胞進行神經分化,在分化第十天可以得到高純度的前腦神經上皮細胞。並在四十五天內我們可以發現唐氏症患者萬能幹細胞分化而來的神經元表現早期阿茲海默症的病徵,包含類澱粉蛋白的堆積、Tau 蛋白的過度磷酸化以及在神經元內表現位置改變。我們將一種當歸的主成分 n-butylidenephthalide (Bdph) 以水膠 pluronic F127 包覆以減低其細胞毒性,並且發現 F127-Bdph 有促進 Wnt 訊息傳遞路徑的活性。在經過 F127-Bdph 處理後我們發現唐氏症神經元分泌至培養液中的類澱粉蛋白以及 Tau 和磷酸化的 Tau 蛋白均有顯著的下降。結合以上結果,我們發現唐氏症萬能幹細胞為一種適當的阿茲海默症細胞模型,並對高效率的藥物篩選有良好的潛力。我們也發現一種新的小分子可能對阿茲海默症的治療有所助益。
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