Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, China.
Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, China.
Anal Chim Acta. 2017 Nov 22;994:1-9. doi: 10.1016/j.aca.2017.09.030. Epub 2017 Sep 28.
Hydrogen sulfide (HS) is a new endogenously generated gasotransmitter and has implicated in many physiologies and pathologies closely related to its intracellular and intercellular signaling transduction. Although many fluorescent probes have been exploited to track and quantify HS in living systems, none of them could be used for monitoring intercellular transmission of HS. Herein, we developed a cell surface specific HS probe, 4-azido-6-sulfo-N-hexadecyl-1,8-naphthalimide, sodium salt (ASNHN-N), trying to investigate the behaviors of extracellular release of HS. ASNHN-N is week fluorescent and could react with HS at 37 °C in pH 7.4 buffer solutions to form product ASNHN-NH with strong fluorescence (Φ = 0.22). Using ASNHN-N as HS probe, excellent linear correlation versus the concentration of NaHS was obtained ranging from 0 to 10 μM and the detection limit was 0.75 μM. With the lipid anchor and the hydrophilic sulfonic group introduced into the 1,8-naphthalimide (a skeleton of two-photon fluorescent probe), the amphiphilic probe is located at the surface of living cells which can record HS efflux from the cell diffusing across the plasma membrane in living cells and deep-tissue by using two-photon microscopy. Thus we present a new strategy for further studying the mechanism of signaling molecules in cell communication and signal pathways.
硫化氢 (HS) 是一种新的内源性气体递质，与其细胞内和细胞间信号转导密切相关，参与了许多生理和病理过程。尽管已经开发出许多荧光探针来跟踪和定量活系统中的 HS，但没有一种探针可用于监测 HS 的细胞间传递。在此，我们开发了一种细胞表面特异性的 HS 探针，4-叠氮基-6-磺酸-N-十六烷基-1,8-萘二甲酰亚胺，钠盐（ASNHN-N），试图研究 HS 细胞外释放的行为。ASNHN-N 荧光较弱，在 37°C 时在 pH 7.4 的缓冲溶液中与 HS 反应，形成强荧光的产物 ASNHN-NH（Φ=0.22）。使用 ASNHN-N 作为 HS 探针，在 0 到 10 μM 的范围内与 NaHS 的浓度呈良好的线性关系，检测限为 0.75 μM。通过在 1,8-萘二甲酰亚胺（双光子荧光探针的骨架）中引入脂锚和亲水性磺酸基，两亲性探针位于活细胞表面，可通过双光子显微镜记录从扩散穿过质膜的活细胞中 HS 的流出。因此，我们提出了一种新策略，用于进一步研究细胞通讯和信号通路中信号分子的机制。
