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目的:观察水杨酸钠(SS)对大鼠耳蜗螺旋神经节神经元(SGN)的多巴胺受体(DR)亚型表达的影响,以及观察SS作用下,DR对SGN的N-甲基-D-天冬氨酸(NMDA)受体和γ-氨基丁酸(GABA)a受体亚单位的影响,探讨SS介导的受体之间的相互作用。方法:应用免疫荧光技术,检测新生SD大鼠SGN中DR两家族DR1、DR2的表达;应用RT-PCR技术,检测SS处理后的SGN中DR1和DR2亚型的mRNA变化,观察在激活和抑制DR活动时SS对SGN的NMDA受体亚单位NR1、GABAa受体亚单位GABRα2的mRNA表达的影响。结果:免疫荧光显示:在SGN的胞体与突起均有DR1、DR2的免疫荧光显示。RT-PCR结果显示:(1)SGN有DR亚型(DRd1~DRd5)、GABRα2、NR1的表达;(2)5mmol/L SS作用SGN 30min后,除DRd3外,其余DR亚型及GABRα2、NR1的mRNA表达量均明显升高,其中DRd1表达增加34.64%(t=-5.123,P=0.007);DRd2表达增加34.60%(t=-5.206,P=0.006);DRd4表达增加20.87%(t=-3.337,P=0.029);DRd5表达增加26.42%(t=-6.054,P=0.004);GABRα2表达增加30.41%(t=-2.839,P=0.047);NR1表达增加39.22%(t=-6.243,P=0.003);(3)SS分别与多巴胺(100μmol/L)、DR1激动剂(SKF38393,20μmol/L)、DR2激动剂(Quinpirole,20μmol/L)分别作用SGN 30min后,GABRα2、NR1的mRNA表达量均明显升高,其中,GABRα2表达分别增加21.78%、27.45%、33.02%、33.42%(F=12.399,P=0.001),NR1表达分别增加28.70%、26.82%、29.03%、35.05%(F=50.395,P=0.000);(4)SS+DR1拮抗剂(SCH23390,20μmol/L)、SS+DR2拮抗剂(Eticlopride,20μmol/L)分别作用SGN 30min后,与单独SS作用相比,GABRα2的mRNA表达分别减少29.56%、37.10%(F=22.101,P=0.000),NR1的mRNA表达分别减少37.62%、32.83%(F=72.933,P=0.000)。结论:SS可诱导SGN的大部分DR亚型的mRNA表达明显升高;SS可能通过DR影响SGN上GABAaR、NMDAR的mRNA表达。
Objective: To observe the effects of sodium salicylate (SS) on the expression of dopamine receptor (DR) in the rat spiral ganglion neurons (SGN) and the effect of DR on the expression of N-methyl- D-aspartate (NMDA) receptor and gamma-aminobutyric acid (GABA) a receptor subunits to explore SS-mediated interactions between receptors. Methods: Immunofluorescence was used to detect the expression of DR1 and DR2 in DRNs of SGNs of SD rats. RT-PCR was used to detect the mRNA expressions of DR1 and DR2 subtypes in SGNs after SS treatment. Effects of SS on the mRNA Expression of NMDA Receptor Subunit NR1 and GABAa Receptor Subunit GABRα2 of SGN in DR Activity. Results: Immunofluorescence showed that there were immunofluorescence staining of DR1 and DR2 in SGN’s somatic cells and protrusions. RT-PCR results showed that: (1) The expression of DR subtype DRD1-DRd5 and GABRα2, NR1 in SGN were observed. (2) After treated with 5mmol / L SS for 30min, (T = -5.123, P = 0.007); the expression of DRd2 increased 34.60% (t = -5.206, P = 0.006); the expression of DRd4 increased by 20.87% (t = -3.337, P = 0.029). The expression of DRd5 increased by 26.42% (t = -6.054, P = 0.004), the expression of GABRα2 increased by 30.41% (t = -2.839, , P = 0.003). (3) After treated with SS for 30min at the concentrations of 100μmol / L, 30μmol / L and DR1 agonists (SKF38393,20μmol / L) and DR2 agonist (20μmol / the expression of GABRα2 increased by 21.78%, 27.45%, 33.02%, 33.42% (F = 12.399, P = 0.001), the expression of NR1 increased by 28.70%, 26.82%, 29.03%, 35.05% (F = 50.395, P = 0.000). (4) SS + DR1 antagonist (SCH23390, 20μmol / L) and SS + DR2 antagonist (Eticlopride, 20μmol / L) The mRNA expression of GABRα2 decreased by 29.56% and 37.10%, respectively (F = 22.101, P = 0.000). The mRNA expression of NR1 decreased by 37.62% and 32%, respectively. 83% (F = 72.933, P = 0.000). Conclusion: SS can induce the expression of most DR subtypes of SGNs significantly. SS may affect the expression of GABAaR and NMDAR mRNA by SGS.