Selective detection of endogenous H₂S in living cells and the mouse hippocampus using a ratiometric fluorescent probe.
Selective detection of endogenous H₂S in living cells and the mouse hippocampus using a ratiometric fluorescent probe.
复制标题
DOI：
10.1038/srep05870
复制
发表时间：
2014-07-29
影响因子：
4.6
通讯作者：
Zhao J
中科院分区：
文献类型：
--
作者：
Zhang L;Meng WQ;Lu L;Xue YS;Li C;Zou F;Liu Y;Zhao J
作为三种气体变送器之一，硫化氢的基本信号作用受到越来越多的关注。需要新的工具来准确检测细胞和组织中的硫化氢，以探索其生物学功能。报道了一种基于对硝基苄基的比率荧光探针RHP-2，它具有检测下限低、选择性高、光稳定性好等特点。发射光强比与PBS缓冲液和牛血清中硫化物浓度呈良好的线性关系。我们的探针被应用于活细胞中内源性硫化氢的比率测定和成像。此外，RHP-2还被用作检测小鼠海马区内源性硫化氢的有效工具。我们观察到，在慢性不可预测的轻度应激诱导的抑郁模型中，小鼠海马区硫化物浓度显著降低，胱硫醚β合成酶(Cbs)基因和cbs蛋白的表达下调。提示内源性硫化氢浓度降低可能参与慢性应激抑郁的发病机制。
As one of three gasotransmitters, the fundamental signalling roles of hydrogen sulphide are receiving increasing attention. New tools for the accurate detection of hydrogen sulphide in cells and tissues are in demand to probe its biological functions. We report the p-nitrobenzyl-based ratiometric fluorescent probe RHP-2, which features a low detection limit, high selectivity and good photostability. The emission intensity ratios had a good linear relationship with the sulphide concentrations in PBS buffer and bovine serum. Our probe was applied to the ratiometric determination and imaging of endogenous H2S in living cells. Furthermore, RHP-2 was used as an effective tool to measure endogenous H2S in the mouse hippocampus. We observed a significant reduction in sulphide concentrations and downregulated expression of cystathionine β-synthetase (CBS) mRNA and CBS protein in the mouse hippocampus in a chronic unpredictable mild stress (CUMS)-induced depression model. These data suggested that decreased concentrations of endogenous H2S may be involved in the pathogenesis of chronic stress depression.