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標題: | 探討NLRP3-Caspase 1 機制在鋅離子引發大鼠初代培養神經元死亡的角色 The Roles of NLRP3-Caspase 1 Pathway in Zn2+-induced Cell Death of Primary-Cultured Rat Embryonic Cortical Neuron |
作者: | You-Hao He 何祐豪 |
指導教授: | 潘建源(Chien-Yuan Pan) |
關鍵字: | 神經退化性疾病,多巴胺能神經細胞,鋅離子,發炎反應,基因表現, Neurodegenerative disease,dopaminergic neurons,Zinc ion,Neuroinflammation,Gene expression, |
出版年 : | 2019 |
學位: | 碩士 |
摘要: | 帕金森氏症(Parkinson’s disease, PD)是非常普遍的神經退化性疾病僅次於阿茲海默症,研究顯示在全世界高於60 歲的人口中有2%的人會罹患PD。而PD的致病機制主要是因為腦部黑質的多巴胺能神經細胞死亡,而造成神經細胞死亡原因很多如氧化壓力、粒線體功能失常、DNA損傷及小膠質細胞的活化等機制。在我們過去的研究中證實了多巴胺會引發細胞自噬及胞質內鋅離子濃度上升最終導致神經細胞死亡。另外過去實驗室透過次世代定序分析發現多巴胺會造成IL-1β及許多與神經發炎反應相關的基因表現量上升,而這些基因都涉及細胞焦亡的途徑但確切機制尚未清楚。為了探究在細胞死亡的過程中鋅離子對於神經發炎反應之間的影響,因此我們使用初代細胞培養的大鼠神經細胞處理多巴胺250 µM 及 氯化鋅 200 µM 18小時引發細胞死亡並測試抑制神經發炎反應對於抵抗細胞死亡的效果。除此之外細胞預處理發炎反應的抑制劑MCC950及VX-765 都可以減少氯化鋅所造成的細胞死亡。從PCR實驗數據中我們發現神經細胞處理多巴胺及氯化鋅後神經發炎相關的基因 IL-1β表現確實增加。因此根據我們的實驗結果可以說明發炎反應參與了鋅離子濃度上升引發的細胞死亡,而此類型的細胞死亡應該是細胞焦亡。我們的發現提供了一個新的關於多巴胺所造成的細胞死亡機制,且在未來可以發展成為神經退化性疾病的療法。 Parkinson’s disease (PD) is the second most common neurodegenerative disease, affecting about 2% of the world’s population over 60 years old. The pathogenic mechanism of PD is the loss of dopaminergic neurons in the substantia nigra (SN), which may be caused by the oxidative stress, mitochondrial dysfunction, DNA damage, microglial cell activation etc. Our previous work has verified that dopamine (DA) can induce autophagy and elevate intracellular Zn2+ concentration resulting in the death of cultured neurons, however, it is not clear the roles Zn2+ plays in these processes. Our preliminary results using NGS analysis shows that DA treatment increase the expression of IL-1β and several neuroinflammation-related genes suggesting the involvement of pyroptosis but not well-characterized yet. It is not clear how Zn2+-neuroinflammation pathway causes the cell death; therefore, we further investigated the roles of inflammasome plays in primary cultured rat embryonic cortical neurons after DA or Zn2+ treatment. We treated the neurons with DA (250 µM) and /or Zn2+ (200 µM) for 18 hrs to induce neuron death and tested the effects of inhibitor against the NLRP3-inflammasome pathway. MCC950, an NLRP3-inflammasome inhibitor, reduced ZnCl2-induced neuron death. VX-765 (caspase 1 inhibitor) can reverse the ZnCl2-induced neuron death. Therefore, these results suggest the involvement of neuroinflammation in Zn2+-induced cell death, which may be mediated by the pyroptosis pathway. The PCR results show that the expression levels of IL-1β increased under the DA and Zn2+ treatment. Our findings provide a new mechanism involved in DA-induced neuron death from which we can develop a new therapeutic strategy to alleviate the neurodegeneration. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/73880 |
DOI: | 10.6342/NTU201903490 |
全文授權: | 有償授權 |
顯示於系所單位: | 生命科學系 |
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