Abstract
Objective
To determine whether activation and/or inhibition of α-adrenoreceptors influences substance P (SP) release from dorsal root ganglion (DRG) primary sensory neurons in vitro.
Methods
DRGs were dissected from 15-day embryonic Wistar rats. DRG neurons were dissociated and cultured for 2 d and then exposed to noradrenaline (NA) alone (1×10−4 mol/L), or along with the α1-adrenoreceptor antagonist prazosin (1×10−6 mol/L) or the α2-adrenoreceptor antagonist yohimbine (1×10−5 mol/L) for 4 d. Then, RT-PCR was used to determine the levels of preprotachykinin (PPT) mRNA encoding for SP and Western blot to assess the protein levels of SP. Basal and capsaicin (CAP)-evoked SP release were measured by enzyme-linked immunosorbent assay (ELISA).
Results
CAP-evoked SP release was sensitized by NA and this effect was inhibited by pre-incubation with prazosin but not with yohimbine. The levels of PPT mRNA, SP peptide, and basal SP release did not change significantly in any of the experimental conditions.
Conclusion
NA may significantly increase CAP-evoked SP release through activation of α-adrenoreceptors, which may contribute to noradrenergic pain modulation.
摘要
目的
观察激活或抑制α-肾上腺素受体是否影响体外培养的背根神经节(dorsal root ganglion, DRG)神经元 P物质(substance P, SP)的释放。
方法
胎龄15天的Wistar大鼠DRG神经元培养2天后, 分别用去甲肾上腺素(noradrenaline, NA)(1×10−4 mol/L)、 α1-受体拮抗剂哌唑嗪(1×10−6 mol/L)+NA(1×10−4 mol/L)、 α2-受体拮抗剂育亨宾(1×10−5 mol/ L)+NA(1×10−4 mol/L)孵育4天。 用RT-PCR法检测DRG神经元编码SP蛋白的PPT mRNA表达水平, 用Western blot 法检测DRG神经元SP蛋白的表达水平, 用酶联免疫吸附测定法检测SP的基础释放量和辣椒素刺激后的释放量。
结果
NA单独孵育显著增加了DRG神经元辣椒素刺激后的SP释放量, α1-受体拮抗剂哌唑嗪预处理可阻断NA的效应, 而α2-受体拮抗剂育亨宾不产生此作用。 在各种实验条件下, PPT mRNA水平、 SP蛋白表达水平和SP的基础释放量没有显著性差异。
结论
NA通过激活α1-受体增加了DRG神经元辣椒素刺激后的SP释放量, 这一作用可能与去甲肾上腺素能的疼痛调节有关。
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Wang, YJ., Li, XF., Ding, F. et al. Noradrenaline regulates substance P release from rat dorsal root ganglion neurons in vitro . Neurosci. Bull. 27, 300–306 (2011). https://doi.org/10.1007/s12264-011-1034-4
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DOI: https://doi.org/10.1007/s12264-011-1034-4