探討親蛋白質腎毒素硫酸吲哚酚誘發斑馬魚之神經退化性影響

Indoxyl sulfate induce neuron degeneration in zebrafish

邱振浩(Chen-Hao Chiu)

指導教授 ： 劉旺達

高雄醫學大學/生命科學院/生物科技學系碩士班/碩士(2019年)

摘要

本研究的目的是使用斑馬魚作為模型來研究硫酸吲哚酚對中樞神經系統細胞的影響並瞭解其機制。本研究使用在神經元專一表現EGFP的基因轉殖斑馬魚品系Tg(HuC：EGFP)來觀察硫酸吲哚酚對其的影響效應。結果顯示，硫酸吲哚酚處理的斑馬魚腦和脊髓中神經元的數量減少。神經元的結構或功能的喪失表示硫酸吲哚酚可能在斑馬魚中誘導神經元退化。通過使用ROS探針染色的檢測方法闡明了硫酸吲哚酚誘導的ROS反應。藉由組織切片染色分析更驗證了受損的結果。此外，通過3D列印裝置來進行硫酸吲哚酚處理的斑馬魚胚胎活動力的影響。我們以Ingenuity Pathway Analysis (IPA)分析經硫酸吲哚酚處理後斑馬魚胚胎的全基因表現。其中，預測活化的途徑有氧化壓力和發炎反應相關的訊息傳導路徑以及NMDA相關傳導途徑受到抑制，此可能與硫酸吲哚酚誘導的神經元損失相關。

關鍵字

硫酸吲哚酚 ； 斑馬魚 ； 神經退化

並列摘要

The purpose of this study is using zebrafish as a model to investigate the mechanism of IS-induced CNS cells lose. In this study, the generated transgenic zebrafish line Tg(HuC:EGFP) that expresses EGFP in neuron was used to exam the IS effect. Results revealed that the number of neuron was decreased in IS-treated fish brain and spinal cord which implied IS may induce neuron degeneration in zebrafish. I clarified IS-induced ROS response by using whole mount ROS-detection method. Thus, neuron damage was verified by histological analysis. Moreover, IS-treated embryos show less activity than the control by locomotor activity test. We identified the global gene expression of IS-treated embryos, and analyzed by Ingenuity Pathway Analysis. In which, activated oxidative stress and inflammatory response signaling pathway, and inhibited NMDA associate pathway were predicted to associate with IS-induced neuron loss.

並列關鍵字

Indoxyl sulfate ； zebrafish ； neuron degeneration

參考文獻

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