The Effects of Resveratrol in 6-OHDA Treated N2a Cells

• Main Authors: LIN, YA-YIN, 林雅茵
• Other Authors: Chiang, Ming-Chang
• Format: Others
• Language: en_US
• Published: 2017
• Online Access: http://ndltd.ncl.edu.tw/handle/ew95yx

碩士 === 輔仁大學 === 生命科學系碩士班 === 105 === In Parkinson’s Disease (PD), the main reason causes PD is the increasing oxidative stress in brain, which producing reactive oxygen species (ROS), inducing pro-apoptosis in neuron cells that might directly and indirectly damage the functions of mitochondria. Gene and environment are two factors that cause PD. Environmental factors include 6-hydroxydopamine (6-OHDA), MPTP, rotenone, etc. In this article, we use 6-OHDA to establish PD cell model in N2a cell line. 6-OHDA is a sort of catecholaminergic neurotoxin that has a similar structure to dopamine and norepinephrine also has high affinity to lots of catecholaminergic plasma membrane. Thus, it could increase oxidative stress inside cells, such as ROS. The above symptoms will directly inhibit electron transport chain and cause cell death. The function of AMP-activated protein kinase (AMPK) in regulating energy, metabolism, and cell regeneration is reported in plenty of papers. AMPK also plays a key role in neurodegenerative disorders due to its anti-oxidant ability by reducing ROS generation. Resveratrol (3,5,4’-trihydroxy-trans-stilbene) is a polyphenolic compound, which is common in berries, grapes, and red wines. Many researches point out that it’s capable of antioxidant and anti-inflammation. And it shows protective effects in cardiovascular diseases and neurodegenerative disease as well. Resveratrol also related to the activation of AMPK; in hence, it’s been regarded as a potential treatment drug. In this article, we want to observe if resveratrol could have some protective effects on 6-OHDA treated N2a cell, such as antioxidant property by active AMPK pathway. At this point, we found that cell viability in pretreated with resveratrol is significant increasing than treated with 6-OHDA only in MTT assay. In TEM results, we could found that mitochondria morphology of N2a cell could be recovered by pre-treat with resveratrol. In DHE results show that ROS rate of combined group is significantly lower than 6-OHDA group. In immunofluorescence, the expression of p-AMPK, SIRT1, PGC-1 and HO-1 in 6-OHDA group is lower than others. In Q-PCR results, HO-1 gene expression in combined groups is higher than 6-OHDA treated group. In 3D culture, the expression rate of p-AMPK and SIRT1 are the same as 2D cell culture. Thus, we suggested that resveratrol have an antioxidant effect on 6-OHDA treated N2a cell.