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强声暴露对巴马小型猪海马细胞内Ca²⁺变化及其信号通道的影响

张艺耀^1,2, 刘淑红¹, 房涛¹, 范明¹, 朱玲玲^1,*

¹军事医学科学院基础医学研究所，北京 100850；²中国人民解放军空军总医院特勤科，北京 100042

摘要

声音对神经系统有重要影响，本研究旨在探讨噪音或高强度声音刺激对神经系统的影响及其机制。将听力正常的巴马小型猪随机分为正常对照组与强声暴露组。强声暴露组的巴马小型猪暴露于中低频强声(900 Hz-142 dB SPL)环境中15 min，暴露结束后即刻分离出海马组织。用Fluo-4探针观察海马组织细胞内Ca²⁺浓度([Ca²⁺]_i)的变化，用real-time PCR和Western blot分别检测Ca²⁺受体、L-型Ca²⁺通道α2/δ1亚基、PKC和PI3K的mRNA和蛋白表达，用DAPI染色法观察细胞核形态变化。结果显示，相对对照组，强声暴露组小型猪海马组织细胞[Ca²⁺]_i明显增加，L-型Ca²⁺通道α2/δ1亚基、PKC和PI3K mRNA表达上调，Ca²⁺受体和PKC蛋白表达显著上调。此外，强声暴露引起海马组织细胞核出现肿胀变形等损伤样改变。以上结果提示，强声暴露可以通过激活海马组织PKC信号通路，引起[Ca²⁺]_i上调，最终导致海马组织内细胞的损伤。本研究结果不仅揭示了强声引起神经损伤的可能机制，同时为防护强声对神经系统造成的损伤提供了新的思路。

关键词： 强声; 巴马小型猪 ; 海马 ; Ca2+ ; 信号通道

分类号：R363.2+1

Change of intracellular Ca²⁺ concentration and related signaling pathway in hippocampal cells after high-intensity sound exposure

ZHANG Yi-Yao^1,2, LIU Shu-Hong¹, FANG Tao¹, Fan Ming¹, ZHU Ling-Ling^1,*

¹The Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China；²The Department of Special Clinical Examination, Air Force General Hospital of PLA, Beijing 100042, China

Abstract

High-intensity sound often leads to the dysfunction and impairment of central nervous system (CNS), but the underlying mechanism is unclear. The present study was aimed to investigate the related mechanisms of CNS lesions in Bama miniature pig model treated with high-intensity sound. The pigs with normal hearing were divided into control and high-intensity sound (900 Hz-142 dB SPL, 15 min) groups. After the treatment, hippocampi were collected immediately. Fluo-4 was used to indicate intracellular Ca²⁺ concentration ([Ca²⁺]_i) change. Real-time PCR and Western blot were used to detect mRNA and protein expressions of calcium-sensing
receptor, L-Ca²⁺ channel α2/δ1 subunit, PKC and PI3K, respectively. DAPI staining was used to identify nuclear features. The result showed that high-intensity sound exposure resulted in significantly swollen cell nucleus and increased [Ca²⁺]_i in hippocampal cells. Compared with control group, high-intensity sound group showed increased levels of PI3K, PKC and L-Ca²⁺ channel α2/δ1 subunit mRNA expressions, as well as up-regulated PKC and calcium-sensing receptor protein expressions. These results suggest that the high-intensity sound activates PKC signaling pathway and induces calcium overload, eventually leads to hippocampal injury, which would supply a novel strategy to prevent nervous system from high-intensity sound-induced injury.

Key words: high-intensity sound; Bama pig ; hippocampus ; Ca2+ ; signaling pathway

收稿日期：2017-02-13　　录用日期：2017-11-06

通讯作者：朱玲玲　　E-mail: linglingzhu@hotmail.com

DOI: 10.13294/j.aps.2017.0081

引用本文：

张艺耀, 刘淑红, 房涛, 范明, 朱玲玲. 强声暴露对巴马小型猪海马细胞内Ca²⁺变化及其信号通道的影响[J]. 生理学报 2017; 69 (6): 737-742.

ZHANG Yi-Yao, LIU Shu-Hong, FANG Tao, Fan Ming, ZHU Ling-Ling. Change of intracellular Ca²⁺ concentration and related signaling pathway in hippocampal cells after high-intensity sound exposure. Acta Physiol Sin 2017; 69 (6): 737-742 (in Chinese with English abstract).