Huang X, Chen H Y, Wei T J, Qin D X, Liu P Q, Ye W H, Su J P
Department of Otolaryngology Head and Neck Surgery, the First Affiliated Hospital of Guangxi Medical University, Nanning, 530021, China.
Department of Otolaryngology Head and Neck Surgery, Qinzhou Second People's Hospital.
Lin Chuang Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2017 Oct 20;31(20):1593-1598. doi: 10.13201/j.issn.1001-1781.2017.20.012.
To investigate mRNA expression of dopamine receptor subtypes in the rat cochlear spiral ganglion neurons (SGN) following exposure to the sodium salicylate. In addition, we observed the effect of sodium salicylate on N methyl-D-aspartic acid (NMDA) receptor subunit NR1 and gamma aminobutyric acid (GABA)a receptor subunit GABRα2 mRNA under the circumstance of DR activation or blocking. Moreover, we also focused on the the interaction between receptors mediated by SS.Immunofluorescence techniques were applied to detect DR (DR1 and DR2) expression in cultured rat SGN. Moreover, RT-PCR was performed to assess NR1 and GABRα2 subunit mRNA.Immunofluorescence images showed co-localization of DR1/DR2 and βⅢ-tubulin protein in SGN bodies and axons. RT-PCR results illustrated that ①DR subtypes of DRd1-5 were detected in the SGN. ②The mRNA expression of all subtypes of DR and GABRα2, NR1 subunits were obviously upregulated except DRd3 after treatment with sodium salicylate. Among them, DRd1 expression increased 34.64%(=-5.123,=0.007),DRd2 expression increased 34.60%(=-5.206,=0.006),DRd4 expression increased 20.87%(=-3.337,=0.029),DRd5 expression increased 26.42%(=-6.054,=0.004),GABRα2 expression increased 30.41%(=-2.839,=0.047),NR1 expression increased 39.22%(=-6.243,=0.003).③After exposure to sodium salicylate (5 mmol/L), dopamine (100 μmol/L), DR1 agonist (SKF38393,20 μmol/L), DR2 agonist (Quinpirole,20 μmol/L), GABRα2 expression increased 21.78%,27.45%,33.02%,33.42% respectively (=12.399,=0.001),and NR1 expression increased 28.70%,26.82%,29.03%,35.05%(=50.395,=0.000) respectively.④Compared with the group of sodium salicylate treatment alone, both sodium salicylate + DR1 antagonist (SCH23390,20 μmol/L) group and sodium salicylate + DR2 antagonists (Eticlopride,20 μmol/L) group had a suppression on GABRα2 and NR1 mRNA expression.GABRα2 mRNA reduced 29.56%,37.10%(=22.101,=0.000) and NR1 mRNA expression decreased 37.62%,32.83% respectively(=72.933,=0.000).Most of the DR subtypes mRNA expression in SGN were increased following exposure to sodium salicylate. DR may be involved in the effect of sodium salicylate on GABAaR and NMDAR mRNA expression.
研究大鼠耳蜗螺旋神经节神经元（SGN）暴露于水杨酸钠后多巴胺受体亚型的mRNA表达。此外，我们观察了在多巴胺受体（DR）激活或阻断的情况下，水杨酸钠对N-甲基-D-天冬氨酸（NMDA）受体亚基NR1和γ-氨基丁酸（GABA）A受体亚基GABRα2 mRNA的影响。此外，我们还关注了水杨酸钠介导的受体之间的相互作用。应用免疫荧光技术检测培养的大鼠SGN中DR（DR1和DR2）的表达。此外，进行逆转录聚合酶链反应（RT-PCR）以评估NR1和GABRα2亚基mRNA。免疫荧光图像显示DR1/DR2与βⅢ-微管蛋白在SGN胞体和轴突中共定位。RT-PCR结果表明：①在SGN中检测到DRd1-5的DR亚型。②水杨酸钠处理后，除DRd3外，DR、GABRα2、NR1亚基所有亚型的mRNA表达均明显上调。其中，DRd1表达增加34.64%（=-5.123，=0.007），DRd2表达增加34.60%（=-5.206，=0.006），DRd4表达增加20.87%（=-3.337，=0.029），DRd5表达增加26.42%（=-6.054，=0.004），GABRα2表达增加30.41%（=-2.839，=0.047），NR1表达增加39.22%（=-6.243，=0.003）。③暴露于水杨酸钠（5 mmol/L）、多巴胺（100 μmol/L）、DR1激动剂（SKF38393，20 μmol/L）、DR2激动剂（喹吡罗，20 μmol/L）后，GABRα2表达分别增加21.78%、27.45%、33.02%、33.42%（=12.399,=0.001），NR1表达分别增加28.70%、26.82%、29.03%、35.05%（=50.395,=0.000）。④与单独水杨酸钠处理组相比，水杨酸钠+DR1拮抗剂（SCH23390，20 μmol/L）组和水杨酸钠+DR2拮抗剂（依替必利，20 μmol/L）组均对GABRα2和NR1 mRNA表达有抑制作用。GABRα2 mRNA分别降低29.56%、37.10%（= 22.101,=0.000），NR1 mRNA表达分别降低37.62%、32.83%（=72.933,=0.000）。暴露于水杨酸钠后，SGN中大多数DR亚型mRNA表达增加。DR可能参与水杨酸钠对GABA A受体和NMDA受体mRNA表达的影响。
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