神经肽Y及其Y1受体在遗传性癫痫大鼠脑内的表达及意义

Expressions and Co-localizations of Neuropeptide Y and Y1 Receptor in Brain of Tremor Rats

目的探讨遗传性癫痫大鼠(TRM)海马和颞叶皮质中神经肽Y(NPY)及其Y1受体(Y1R)的表达和分布。方法逆转录聚合酶链反应(RT-PCR)检测NPY和Y1R m RNA表达。ELISA试剂盒法检测TRM和Wistar大鼠海马和颞叶皮质中NPY浓度。Western blot检测Y1R蛋白表达。免疫荧光双标记法分析TRM及Wistar大鼠海马CA1、CA3和DG区以及颞叶皮质中NPY与Y1R的分布和共定位。结果 ELISA和RT-PCR结果均显示,TRM海马和颞叶皮质中NPY表达明显上调,与Wistar大鼠比较差异有统计学意义(P<0.01)。Western blot结果证实,TRM海马中Y1R蛋白表达明显低于Wistar大鼠,而在颞叶皮质中显著高于Wistar大鼠。免疫荧光分析发现NPY与Y1R在TRM海马CA1、CA3区神经元细胞和DG区颗粒细胞以及颞叶皮质神经元细胞中分布广泛,并存在共定位且主要定位在细胞膜上。结论在TRM海马和颞叶皮质中,NPY及其Y1R表达出现异常变化,而这种异常表达可能与TRM癫痫发生机制有关。

Objective To explore the expression and distribution of neuropeptide Y （NPY） and Y1 receptor （Y1R） in brain of Tremor rats （TRM）. Methods The mRNA expression of NPY and Y1R was detected by RT-PCR. The concentration of NPY was detected by ELISA kit. The protein expression of Y1R was determined by Western blot in TRM hippocampus and temporal lobe cortex. The distribution and co-localization of these two proteins were analyzed using double-labeling immunofluorescence staining in temporal lobe cortex and hippocampal CA 1, CA3, and DG. Results Compared with Wistar rats, increased NPY and decreased Y1R were observed in the hippocampus of TRM. However, both NPY and Y1R were up-regulated in TRM temporal lobe cortex. NPY and YIR were present and co-localized throughout TRM brains, especially localized on the membranes of pyramidal cells and granule cells. Conclusion The altered expressions of NPY and Y1R were observed in TRM hippocampus and temporal lobe cortex, which may be involved in the generation of TRM epileptiform activity.