Safronova Valentina G, Vulfius Catherine A, Shelukhina Irina V, Mal'tseva Valentina N, Berezhnov Alexey V, Fedotova Eugeniya I, Miftahova Regina G, Kryukova Elena V, Grinevich Andrey A, Tsetlin Victor I
Institute of Cell Biophysics, Russian Academy of Sciences, Institutskaya str., 3, Pushchino, Moscow region, 142290, Russia.
Institute of Cell Biophysics, Russian Academy of Sciences, Institutskaya str., 3, Pushchino, Moscow region, 142290, Russia.
Immunobiology. 2016 Jul;221(7):761-72. doi: 10.1016/j.imbio.2016.01.016. Epub 2016 Mar 4.
Participation of nicotinic acetylcholine receptors (nAChRs) in functioning of polymorphonuclear neutrophils (PMNs) isolated from inflammatory site of mice and expression of different nAChR subunits were studied. Nicotine and acetylcholine (ACh) modified respiratory burst induced by a chemotactic peptide N-formyl-MLF in neutrophils of male (but not female) mice. Antagonists of nAChRs α-cobratoxin (αCTX), α-conotoxins MII and [A10L]PnIA at concentrations of 0.01-5μM, 0.2μM and 1μM, respectively, eliminated nAChR agonist effects. ACh also affected adhesion of PMNs, this effect was also prevented by αCTX (100nM) and MII (1nM). Neutrophils of female mice after chronic nicotine consumption acquired sensitivity to nAChR agonists. Changes of free intracellular Ca(2+) concentration in neutrophils under the action of nAChR ligands were analyzed. In cells with no Ca(2+) oscillations and relatively low resting level of intracellular Ca(2+), nicotine triggered Ca(2+)-spikes, the lag of the response shortened with increasing nicotine concentration. A nicotinic antagonist caramiphen strongly decreased the effect of nicotine. RT-PCR analysis revealed mRNAs of α2, α3, α4, α5, α6, α7, α9, β2, β3, and β4 nAChR subunits. Specific binding of [(125)I]-α-bungarotoxin was demonstrated. Thus in view of the effects and binding characteristics the results obtained suggest a regulatory role of α7, α3β2 or α6* nAChR types in specific functions of PMNs.
研究了烟碱型乙酰胆碱受体（nAChRs）在从小鼠炎症部位分离的多形核中性粒细胞（PMNs）功能中的作用以及不同nAChR亚基的表达。尼古丁和乙酰胆碱（ACh）改变了趋化肽N-甲酰甲硫氨酰-亮氨酰-苯丙氨酸（N-formyl-MLF）诱导的雄性（而非雌性）小鼠中性粒细胞的呼吸爆发。nAChRs拮抗剂α-银环蛇毒素（αCTX）、α-芋螺毒素MII和[A10L]PnIA的浓度分别为0.01 - 5μM、0.2μM和1μM时，消除了nAChR激动剂的作用。ACh还影响PMNs的黏附，这种作用也被αCTX（100nM）和MII（1nM）所阻止。长期摄入尼古丁后，雌性小鼠的中性粒细胞对nAChR激动剂产生了敏感性。分析了nAChR配体作用下中性粒细胞内游离钙离子浓度的变化。在没有钙离子振荡且细胞内钙离子静息水平相对较低的细胞中，尼古丁引发钙离子尖峰，随着尼古丁浓度的增加，反应延迟缩短。烟碱拮抗剂卡拉美芬强烈降低了尼古丁的作用。逆转录聚合酶链反应（RT-PCR）分析显示了α2、α3、α4、α5、α6、α7、α9、β2、β3和β4 nAChR亚基的信使核糖核酸（mRNAs）。证实了[(125)I]-α-银环蛇毒素的特异性结合。因此，鉴于这些作用和结合特性，所得结果表明α7、α3β2或α6* nAChR类型在PMNs的特定功能中具有调节作用。
