Nrf2與神經退化性疾病：啟動子多型性與 以氧化壓力為目標的治療策略

NFE2-related factor 2 (Nrf2) in neurodegenerative disease: promoter polymorphism and therapeutic strategy targeting oxidative stress

巫逸琦(Yi-Ci Wu)

指導教授 ： 李桂楨

國立臺灣師範大學/理學院/生命科學研究所/碩士(2012年)

摘要

Nuclear factor-erythroid 2 (NF-E2)-related factor 2 (Nrf2)是basic leucine zipper (bZIP)的轉錄因子，可調節抗氧化路徑基因的活化，並維持細胞內氧化還原的平衡。氧化壓力的增加與許多神經性退化性疾病有相關性。舉例來說，帕金森氏症(PD)的致病機制就會受到氧化壓力的影響，增強Nrf2的活化會對於神經細胞有保護性的功效。在polyQ不正常擴增的脊髓小腦萎縮症中，擴增的CAG三核苷酸轉譯出的polyQ蛋白，聚集在細胞內增加細胞內的氧化壓力。本論文先以PCR-RFLP技術，對Nrf2基因啟動子-653 A/G、-651 G/A和-617 C/A多型性，進行帕金森氏症患者(n = 480)與年齡、性別配合的正常人(n = 526)的病例-對照組研究。結果多型性基因型、等位基因頻率、單套型等均並未發現相關性。另外，本研究建立誘導式ATXN3/Q14~75的Flp-In SH-SY5Y細胞株，添加retinoic acid誘導分化後，表現具有病理特徵的ATXN3/Q75融合蛋白會形成聚集，並伴隨神經纖維生長減緩。藉高通量影像分析及免疫轉漬分析，此細胞株處理中草藥NH004、NH008、NH021及NH008某組成份衍生物NH008-1可活化Nrf2的表現，並降低ATXN3/Q75融合蛋白的聚集，故可能發展為有潛能的治療策略。

關鍵字

Nrf2 ； 氧化壓力 ； 聚集 ； 多型性 ； ATXN3

並列摘要

Nuclear factor-erythroid 2 (NF-E2)-related factor 2 (Nrf2) is a member of the basic leucine zipper transcription factors that regulate the expression of many antioxidant pathway genes and maintains cellular redox homeostasis. Increased oxidative stress is involved in the pathogenesis of many neurodegenerative diseases. For example, oxidative stress has been implicated as a major contributing factor in Parkinson’s disease (PD) and varying efficiency in the oxidative protection by Nrf2 may influence PD pathogenesis. In polyQ-mediated spinocerebellar ataxias, the expansions of translated CAG repeats in the disease genes result in long polyQ tracts in the respective proteins, leading to accumulation of aggregated polyQ proteins and increased oxidative stress. In this study, PCR-RFLP test was developed to examine the frequency of Nrf2 -653 A/G, -651 G/A and -617 C/A promoter polymorphisms in a larger cohort of PD (n = 480, 49.2% female, age at onset 61.8±11.2 years) and age- and gender-matched controls (n = 526, 50.5% female, age 60.3±13.1 years). No association between polymorphic genotype, allele or haplotype and PD was observed. In addition, Flp-In SH-SY5Y cells with ATXN3/Q14~75 expression in an inducible fashion were established. In retinoic acid-induced differentiated SH-SY5Y cells, the expressed ATXN3/Q75 formed aggregates, accompanying with reducing neurite outgrowth. By combining high content image analysis and immunoblotting, treatment of ATXN3/Q75 cells with Chinese herbs NH004, NH008, NH021 and NH008 derivative NH008-1 activate Nrf2 expression, accompanying decreasing ATXN3/Q75 aggregates. Thus NH004, NH008, NH021 and NH008-1 may be potential therapeutic strategies for polyQ-mediated disease.

並列關鍵字

Nrf2 ； oxidative stress ； aggregate ； polymorphism ； ATXN3

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