富氫水對抗乙型類澱粉胜肽誘發神經毒性之神經保護作用

阿茲海默症是一種腦部退化性疾病。許多證據認為乙型類澱粉胜肽（amyloid-beta peptide；Aβ）會誘發活性氧（reactive oxygen species；ROS）上升造成氧化壓力，導致神經細胞死亡。另外，AMPK、SIRT1與FOXO3a是與抗老化相關的ROS調控蛋白。近來研究認為富氫水（hydrogen-rich water；HW）中大量氫分子具有抗氧化能力。但對於HW抑制Aβ誘發神經細胞死亡的作用機轉尚不清楚。本研究目的在於調查HW能否對抗Aβ誘發的神經細胞死亡，並探討其作用機轉。首先，評估Aβ誘發的神經毒性，發現在Aβ處理下的SK-N-MC神經細胞存活率隨Aβ濃度上升而下降。然後，以超微弱化學發光法測量HW的ROS清除能力，發現HW具有清除H2O2的能力。接著，觀察神經細胞凋亡現象，發現Aβ活化cleavage Caspase3與PARP，誘發神經凋亡，HW則抑制它們活化，減少凋亡。此外，HW活化AMPK、SIRT1與FOXO3a，並減少細胞核中PSer253-FOXO3a的表現。HW對於LPS誘發EOC13.31神經膠細胞發炎反應具有抑制IL-1β與TNF-α表現的能力。因此，本研究認為HW透過掃除H2O2與活化AMPK與SIRT1並抑制FOXO3a而對抗Aβ誘發的神經凋亡，除此，抑制神經發炎反應，達到神經保護作用。

Alzheimer’s disease (AD) is a neurodegenerative disorder. Several lines of evidences suggest amyloid-beta peptides (Aβ) induced reactive oxygen species (ROS) leading neuronal cell death. Additionally, AMP-activated protein kinase (AMPK), Silence information regulator 1 (SIRT1) and Forkhead box protein O3a (FOXO3a) are new important signaling molecules factors which are ROS- modulating proteins, highly related to anti-ageing. In recent studies, showed hydrogen-rich water (HW), containing a huge amount of molecular hydrogen, known to be ROS scavenger, might also carry an antioxidant property. However, the mechanism of how HW inhibits cell death induced by Aβ still unclear. The aim of this study is to investigate whether HW protects against Aβ-induced cell death and the possible mechanisms. In this study, we first investigated the neurotoxicity induced by Aβ, and found the cell viability decrease as the concentration of Aβ increase. Then, we measured the ROS scavenge ability of HW, and discovered an ability of H2O2 scavenging by HW. Next we observed the apoptosis phenomenon; we found cleavage caspase3 and PARP activated by Aβ, and the inhibition by HW. Moreover, we observed the activation of AMPK, SIRT1 and FOXO3a, and PSer253-FOXO3a was decreased in the nucleus by treated with HW. Besides, HW inhibited the expression of IL-1β and TNF-α induced by LPS in EOC13.31 cells. We suggest that the neuroprotective effects of HW against Aβ-induced cell death may be by scavenging H2O2 and activation of AMPK and SIRT1, besides inactivation of FOXO3a, and also inhibition of inflammation.