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目的探讨海马细胞胞浆视黄酸受体(retinoic acid receptor,RAR)α蛋白表达对大鼠空间学习记忆功能的影响。方法 SD大鼠采用完全随机化分组方法分为Morris水迷宫(Morris water maze,MWM)训练组(n=6)和未训练组(n=6),检测海马组织RARα和GluR1表达水平。分别将AAV-control sh RNA和AAV-RARαsh RNAs海马定位注射SD大鼠后,检测其学习记忆功能及海马组织RARα和GluR1表达水平变化。体外利用神经生长因子(neuron growth factor,NGF)诱导PC12细胞成熟分化,检测RARα和GluR1表达水平、胞内钙活性及静息膜电位。结果水迷宫训练组大鼠海马组织RARα和GluR1总蛋白和胞浆蛋白表达水平均较未训练组显著增加(P<0.01)。海马定位注射2周后,Morris水迷宫检测发现AAV-RARαsh RNAs组大鼠学习记忆功能较对照组明显减弱(P<0.05),同时海马RARα和GluR1总蛋白和胞浆蛋白表达水平也明显下降(P<0.01)。经NGF诱导后的PC12细胞静息膜电位更接近于神经元静息膜电位(P<0.01),RARα和GluR1总蛋白和胞浆蛋白表达水平较对照组明显增高(P<0.01),胞内钙活性明显增强(P<0.05)。结论海马细胞胞浆RARα蛋白表达与学习记忆能力呈正相关,在调节突触可塑性中发挥重要作用。
Objective To investigate the effect of retinal acid receptor (RAR) α protein expression on spatial learning and memory in rats. Methods SD rats were randomly divided into Morris water maze training group (n = 6) and untreated group (n = 6), and the expression of RARα and GluR1 in the hippocampus were detected. AAV-control sh RNA and AAV-RARαsh RNAs were respectively injected into the hippocampus of SD rats, and their learning and memory abilities and the expression of RARα and GluR1 in hippocampus were detected. In vitro, neuronal growth factor (NGF) was used to induce the differentiation of PC12 cells. The expression of RARα and GluR1, intracellular calcium activity and resting membrane potential were detected. Results The levels of total protein and cytoplasmic protein of RARα and GluR1 in hippocampus of water maze training group were significantly higher than those of untrained group (P <0.01). Morris water maze test showed that the learning and memory function of AAV-RARαsh RNAs group was significantly weaker than that of the control group (P <0.05), meanwhile the total protein and cytoplasmic protein expression of RARα and GluR1 in the hippocampus were significantly decreased P <0.01). The resting membrane potential of PC12 cells induced by NGF was closer to the resting membrane potential of neurons (P <0.01), and the total protein and cytoplasmic protein expression of RARα and GluR1 were significantly higher than that of the control group (P <0.01) Calcium activity was significantly increased (P <0.05). Conclusion The expression of RARα protein in hippocampus is positively correlated with learning and memory ability, and play an important role in the regulation of synaptic plasticity.