Azoreductase and Target Simultaneously Activated Fluorescent Monitoring for Cytochrome c Release under Hypoxia
Azoreductase and Target Simultaneously Activated Fluorescent Monitoring for Cytochrome c Release under Hypoxia
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偶氮还原酶和靶标同时激活荧光监测缺氧条件下细胞色素 c 的释放
DOI：
10.1021/acs.analchem.8b00554
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发表时间：
2018
影响因子：
7.4
通讯作者：
Yang Ronghua
中科院分区：
文献类型：
--
作者：
Tang Jianru;Huang Caixia;Shu Jinyong;Zheng Jing;Ma D;an;Li Jishan;Yang Ronghua
低氧诱导的细胞凋亡与退行性疾病、自身免疫性疾病和肿瘤疾病密切相关。在细胞凋亡的过程中，细胞色素c(Cytc)的释放被认为是内在途径的关键因素。最近提出了基于亚细胞定位的跟踪活细胞中细胞色素c释放的策略。然而，它们在区分低氧诱导的细胞凋亡过程中细胞色素c的释放与其他刺激过程中缺乏特异性。本文提出了一种将金纳米颗粒(AuNPs)和细胞色素c靶向适配子组成的双链DNA杂交复合体(DSDHC)整合在一起的偶氮还原酶和靶向同时激活的荧光适配子纳米传感器。值得注意的是，DSDHC上标记的偶氮苯部分的使用首先确保了适体纳米传感器可以连接到AuNPs的表面，然后被缺氧相关的偶氮还原酶特异性地还原。线粒体在低氧条件下释放细胞色素c后，细胞色素c的竞争性置换激活了适体纳米传感器的荧光，荧光增强主要依赖于细胞色素c的释放。受此启发，提出了一种新的缺氧条件下细胞色素c的定量分析和动态成像策略。对低氧条件下Cytc释放动态的高空间分辨率监测将为了解低氧条件下的细胞凋亡机制提供一个潜在的丰富机会，从而进一步促进低氧环境下的风险评估和风险降低。
Hypoxia-induced cell apoptosis is closely related to degenerative diseases, autoimmune disorders, and tumor disease. In the process of apoptosis, the release of cytochrome c (Cyt c) is deemed to be a critical factor of the intrinsic pathway. Strategies for tracking Cyt c release in living cells based on the subcellular localization have been proposed recently. However, they are inherently lack of specificity for distinguishing the release of Cyt c in apoptotic process induced by hypoxia from other stimulus. In this paper, an azoreductase and target simultaneously activated fluorescent aptameric nanosensor integrating gold nanoparticles (AuNPs) and Cyt c-targeted aptamer-consisted double-stranded DNA hybridization complex (DSDHC) was proposed. It is worth noting that the employment of azobenzene moiety labeled on the DSDHC first ensured the aptameric nanosensor could be conjugated to the surface of AuNPs and then specifically reduced by hypoxia-related azoreductase. Upon Cyt c released from mitochondrion under hypoxia, the competitive displacement of Cyt c subsequently activated the fluorescence of the aptameric nanosensor and the fluorescence enhancement depended principally on the content of Cyt c release. Inspired by this, a new strategy for quantitative analysis andin situimaging of Cyt c under hypoxic condition was proposed. The high spatial resolution monitoring of the dynamics of Cyt c release under hypoxia will offer a potentially rich opportunity to understand the apoptotic mechanism under hypoxic conditions, thus further facilitating risk assessment and risk reduction for hypoxic environments.