丁酸盐可增强辣椒素诱导的原代培养大鼠背根神经节神经元中P物质和降钙素基因相关肽的释放。

Butyrate sensitizes the release of substance P and calcitonin gene-related peptide evoked by capsaicin from primary cultured rat dorsal root ganglion neurons.

作者信息

Xing Yi, Liu Zhen, Wang Li-Hong, Huang Fei, Wang Huai-Jing, Li Zhen-Zhong

机构信息

Department of Anatomy, Shandong University School of Medicine, Jinan 250012, China.

出版信息

Neuro Endocrinol Lett. 2006 Dec;27(6):695-701.

PMID:17187013

Abstract

OBJECTIVES

To investigate whether butyrate increases substance P (SP) and calcitonin gene-related peptide (CGRP) release evoked by capsaicin from primary cultured dorsal root ganglion (DRG) neurons.

METHODS

DRG was dissected out from embryonic 15-day-old Wistar rat and cultured as dissociate cells for 24 h then exposed to butyrate (0.01 mmol/L, 0.1 mmol/L, 1 mmol/L, 10 mmol/L, respectively) for another 48 h. The neurons cultured continuously in media served as normal control. All above cultured samples were processed for detecting expression of mRNA for SP, CGRP and vanilloid receptor 1 (VR1) of DRG neurons by RT-PCR, and VR1 protein expression by Western blot. SP and CGRP basal release levels were measured by radioimmunoassay (RIA). After that, the DRG cells for RIA were stimulated by capsaicin (300 nmol/L) for 5 min and the culture media were harvested for detecting SP and CGRP release levels by RIA. The neurons exposed to vehicle solution served as vehicle controls.

RESULTS

Exposure of butyrate on DRG neurons at higher concentrations (1 mmol/L, 10 mmol/L) for 48 h increased expression mRNA for SP and CGRP than that at lower concentrations (0.01 mmol/L, 0.1 mmol/L) and normal control (P<0.001). VR1 mRNA and VR1 protein expression increased in a dose-dependent manner after exposure of different concentrations of butyrate. Butyrate did not alter the basal release, significantly enhanced neuropeptide release evoked by capsaicin. SP and CGRP release levels in the culture media exposed by butyrate at higher concentrations (1 mmol/L, 10 mmol/L) for 48 h and then stimulated by capsaicin were higher than that at lower concentrations (0.01 mmol/L, 0.1 mmol/L) (P<0.001). The exposure of vehicle solution did not produce any increase of SP and CGRP release from primary cultured DRG neurons.

DISCUSSION

Butyrate may promote the expression of mRNA for SP, CGRP and increase sensitivity of capsaicin on SP and CGRP release from primary cultured rat dorsal root ganglion neurons. The promotion of VR1 mRNA and VR1 protein expression by butyrate implicated that VR1 may be involved in the mechanisms of sensory neuropeptide release evoked by capsaicin.

摘要

目的

研究丁酸盐是否能增加辣椒素诱导的原代培养背根神经节（DRG）神经元中P物质（SP）和降钙素基因相关肽（CGRP）的释放。

方法

从15日龄胚胎Wistar大鼠中取出DRG，培养为解离细胞24小时，然后分别用丁酸盐（0.01 mmol/L、0.1 mmol/L、1 mmol/L、10 mmol/L）处理48小时。持续在培养基中培养的神经元作为正常对照。对上述所有培养样本进行处理，通过逆转录聚合酶链反应（RT-PCR）检测DRG神经元中SP、CGRP和香草酸受体1（VR1）的mRNA表达，通过蛋白质印迹法检测VR1蛋白表达。通过放射免疫测定法（RIA）测量SP和CGRP的基础释放水平。之后，用辣椒素（300 nmol/L）刺激用于RIA的DRG细胞5分钟，收集培养基，通过RIA检测SP和CGRP的释放水平。暴露于溶剂溶液的神经元作为溶剂对照。

结果

较高浓度（1 mmol/L、10 mmol/L）的丁酸盐处理DRG神经元48小时后，SP和CGRP的mRNA表达高于较低浓度（0.01 mmol/L、0.1 mmol/L）和正常对照（P<0.001）。不同浓度丁酸盐处理后，VR1 mRNA和VR1蛋白表达呈剂量依赖性增加。丁酸盐不改变基础释放，但显著增强辣椒素诱导的神经肽释放。较高浓度（1 mmol/L、10 mmol/L）的丁酸盐处理DRG神经元48小时后再用辣椒素刺激，培养基中SP和CGRP的释放水平高于较低浓度（0.01 mmol/L、0.1 mmol/L）（P<0.001）。暴露于溶剂溶液未使原代培养的DRG神经元中SP和CGRP的释放增加。