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用于快速监测小鼠心房内源性乙酰胆碱释放的发光纳米颗粒。

Luminescent nanoparticles for rapid monitoring of endogenous acetylcholine release in mice atria.

作者信息

Mukhametshina Alsu, Petrov Alexey, Fedorenko Svetlana, Petrov Konstantin, Nizameev Irek, Mustafina Asiya, Sinyashin Oleg

机构信息

A. E. Arbuzov Institute of Organic and Physical Chemistry, Kazan Scientific Center, Russian Academy of Sciences, Kazan, Russian Federation.

Department of Normal Physiology, Kazan State Medial University, Kazan, Russian Federation.

出版信息

Luminescence. 2018 May;33(3):588-593. doi: 10.1002/bio.3450. Epub 2018 Jan 29.

Abstract

The present work introduces for the first time a nanoparticulate approach for ex vivo monitoring of acetylcholinesterase-catalyzed hydrolysis of endogenous acetylcholine released from nerve varicosities in mice atria. Amino-modified 20-nm size silica nanoparticles (SNs) doped by luminescent Tb(III) complexes were applied as the nanosensors. Their sensing capacity results from the decreased intensity of Tb(III)-centred luminescence due to the quenching effect of acetic acid derived from acetylcholinesterase-catalyzed hydrolysis of acetylcholine. Sensitivity of the SNs in monitoring acetylcholine hydrolysis was confirmed by in vitro experiments. Isolated atria were exposed to the nanosensors for 10 min to stain cell membranes. Acetylcholine hydrolysis was monitored optically in the atria samples by measuring quenching of Tb(III)-centred luminescence by acetic acid derived from endogenous acetylcholine due to its acetylcholinesterase-catalyzed hydrolysis. The reliability of the sensing was demonstrated by the quenching effect of exogenous acetylcholine added to the bath solution. Additionally, no luminescence quenching occurred when the atria were pre-treated with the acetylcholinesterase inhibitor paraoxon.

摘要

本研究首次介绍了一种纳米颗粒方法,用于体外监测小鼠心房神经膨体释放的内源性乙酰胆碱的乙酰胆碱酯酶催化水解。掺杂有发光Tb(III)配合物的氨基修饰的20纳米大小的二氧化硅纳米颗粒(SNs)被用作纳米传感器。它们的传感能力源于由于乙酰胆碱酯酶催化乙酰胆碱水解产生的乙酸的猝灭作用,导致以Tb(III)为中心的发光强度降低。通过体外实验证实了SNs监测乙酰胆碱水解的灵敏度。分离的心房暴露于纳米传感器10分钟以对细胞膜进行染色。通过测量由于内源性乙酰胆碱的乙酰胆碱酯酶催化水解产生的乙酸对以Tb(III)为中心的发光的猝灭,对心房样品中的乙酰胆碱水解进行光学监测。添加到浴液中的外源性乙酰胆碱的猝灭作用证明了传感的可靠性。此外,当心房用乙酰胆碱酯酶抑制剂对氧磷预处理时,未发生发光猝灭。

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