烟碱对大鼠海马脑片β_1-肾上腺素能受体的影响

Effect of Nicotine on β1-adrenergic Receptor Gene Expression and Protein Level in Rat Hippocampal Slice

[目的]研究烟碱对β1-肾上腺素能受体的作用。[方法]将400μm厚的海马脑片置于预先经体积分数95％O2和5％CO2饱和的28℃人工脑脊液中孵育120 min后加入烟碱,终浓度为10μmol/L,分别于烟碱作用后30,60,90,120min,用半定量逆转录-聚合酶链反应和免疫印迹法检测大鼠海马脑片中β1-肾上腺素能受体mR-NA表达和蛋白水平的变化。[结果]大鼠海马脑片经烟碱作用后,β1-肾上腺素能受体mRNA表达明显增加,在60min时最为显著(P<0.05);蛋白水平的变化与基因表达变化有较好的一致性,但蛋白变化的高峰出现时间比mR-NA晚,且变化幅度更大。[结论]烟碱的急性刺激能使β1-AR肾上腺素能受体基因转录和蛋白表达上调,其变化规律提示烟碱不仅调控β1-肾上腺素能受体基因转录,同时也对β1-肾上腺素能受体转录后进行调控,最终导致β1-肾上腺素能受体蛋白水平升高。

[Objective] To investigate the effect of nicotine on β1-adrenergic receptor in hippocampal slice. [Methods] Hippocampal slices(400 μm thick) were incubated in artificial cerebrospinal fluid (ACSF) previously saturated with 95% O2 and 5% CO2 at 28 ℃ for 120 min, and then 10 μmol/L of nicotine was added. 30, 60, 90, 120 mins after addition of nicotine, mRNA of β1-adrenergic receptor of these samples was studied with semiquantitative reverse transcription-polymerase chain reaction (RT-PCR), and the protein level was studied with Western blot. [Results] The mRNA gene expression and the protein level of β1-adrenergic receptor in hippocampal slices were increased during treatment with nicotine. The increasing peak of protein level was later but higher than that of mRNA level. [Conclusion] Both expression of β1- adrenergic receptor gene transcription and post-transcriptional protein level in rat hippocampus are altered by nicotine.