Matsushita Yumi, Manabe Miki, Kitamura Naoki, Shibuya Izumi
Laboratory of Veterinary Physiology, Faculty of Agriculture, Tottori University, Tottori, Japan.
PLoS One. 2018 Jan 5;13(1):e0191032. doi: 10.1371/journal.pone.0191032. eCollection 2018.
Transient receptor potential vanilloid type 1 (TRPV1) is a polymodal receptor channel that responds to multiple types of stimuli, such as heat, acid, mechanical pressure and some vanilloids. Capsaicin is the most commonly used vanilloid to stimulate TRPV1. TRPV1 channels are expressed in dorsal root ganglion neurons that extend to Aδ- and C-fibers and have a role in the transduction of noxious inputs to the skin into the electrical signals of the sensory nerve. Although noradrenergic nervous systems, including the descending antinociceptive system and the sympathetic nervous system, are known to modulate pain sensation, the functional association between TRPV1 and noradrenaline in primary sensory neurons has rarely been examined. In the present study, we examined the effects of noradrenaline on capsaicin-evoked currents in cultured dorsal root ganglion neurons of the rat by the whole-cell voltage clamp method. Noradrenaline at concentrations higher than 0.1 pM significantly reduced the amplitudes of the inward capsaicin currents recorded at -60 mV holding potential. This inhibitory action was reversed by either yohimbine (an α2 antagonist, 10 nM) or propranolol (a β antagonist, 10 nM). The α2 agonists, clonidine (1 pM) and dexmedetomidine (1 pM) inhibited capsaicin currents, and yohimbine (1 nM) reversed the effects of clonidine. The inhibitory action of noradrenaline was not seen in the neurons pretreated with pertussis toxin (100 μg/ml for 24 h) and the neurons dialyzed intracellularly with guanosine 5'- [β-thio] diphosphate (GDPβS, 200 μM), the catalytic subunit of protein kinase A (250 U/ml) or okadaic acid (1 μM). These results suggest that noradrenaline directly acts on dorsal root ganglion neurons to inhibit the activity of TRPV1 depending on the activation of α2-adrenoceptors followed by the inhibition of the adenylate cyclase/cAMP/protein kinase A pathway.
瞬时受体电位香草酸亚型1(TRPV1)是一种多模态受体通道,可对多种类型的刺激产生反应,如热、酸、机械压力和一些香草酸类物质。辣椒素是最常用于刺激TRPV1的香草酸类物质。TRPV1通道表达于延伸至Aδ纤维和C纤维的背根神经节神经元中,在将皮肤的有害刺激转化为感觉神经的电信号过程中发挥作用。虽然已知包括下行镇痛系统和交感神经系统在内的去甲肾上腺素能神经系统可调节痛觉,但TRPV1与去甲肾上腺素在初级感觉神经元中的功能关联很少被研究。在本研究中,我们采用全细胞膜片钳方法研究了去甲肾上腺素对大鼠培养的背根神经节神经元中辣椒素诱发电流的影响。浓度高于0.1 pM的去甲肾上腺素显著降低了在-60 mV钳制电位下记录的内向辣椒素电流的幅度。这种抑制作用可被育亨宾(一种α2拮抗剂,10 nM)或普萘洛尔(一种β拮抗剂,10 nM)逆转。α2激动剂可乐定(1 pM)和右美托咪定(1 pM)抑制辣椒素电流,而育亨宾(1 nM)可逆转可乐定的作用。在用百日咳毒素(100 μg/ml处理24小时)预处理的神经元以及用鸟苷5'-[β-硫代]二磷酸(GDPβS,200 μM)、蛋白激酶A催化亚基(250 U/ml)或冈田酸(1 μM)进行细胞内透析的神经元中,未观察到去甲肾上腺素的抑制作用。这些结果表明,去甲肾上腺素直接作用于背根神经节神经元,通过激活α2肾上腺素能受体,随后抑制腺苷酸环化酶/cAMP/蛋白激酶A途径来抑制TRPV1的活性。