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目的探讨神经肽Y(NPY)对原代小神经胶质细胞生物活性及生成IL-1β的影响。方法培养的原代大鼠皮层小胶质细胞,将细胞分为Control组、LPS组、NPY+LPS组、NPY组和BIBP3226+NPY+LPS组,每组3个样本,培养各组细胞6h。经免疫细胞化学荧光染色后,显微镜下观察小胶质细胞的形态学变化。Elisa方法检测培养液中IL-1β蛋白含量,RT-PCR方法检测小胶质细胞中IL-1βm RNA表达水平。结果孵育各组小胶质细胞6h后,LPS组培养液中IL-1β蛋白的含量及细胞中IL-1βm RNA表达水平分别为(961.00±83.50)pg/m L和5.59±0.87,显著高于Control组的96.33±24.58 pg/m L和1.05±0.12(P<0.05),小胶质细胞处于活化状态;LPS+NPY组IL-1β蛋白含量和m RNA表达水平分别为(411.33±55.00)pg/m L和1.93±0.45,与LPS组相比显著降低(P<0.05),小胶质细胞活化水平降低;IBP3226+NPY+LPS组IL-1β蛋白含量和m RNA表达水平分别为(886.00±97.53)pg/m L和4.51±0.71,与LPS+NPY组相比显著增高(P<0.05);LPS组和IBP3226+NPY+LPS组之间无统计学意义。NPY组与对照组无统计学意义。结论 NPY通过作用于NPY Y1受体降低小神经胶质细胞的生物活性,抑制其生成IL-1β。
Objective To investigate the effect of neuropeptide Y (NPY) on the biological activity of primary microglia and the production of IL-1β. Methods Primary rat cortical microglial cells were cultured and divided into control group, LPS group, NPY + LPS group, NPY group and BIBP3226 + NPY + LPS group, with 3 samples in each group. The cells in each group were cultured for 6 hours. After immunocytochemical staining, the morphological changes of microglia were observed under the microscope. Elisa method was used to detect the content of IL-1βin culture medium, and the level of IL-1βmRNA in microglia was detected by RT-PCR. Results After incubation for 6h, the levels of IL-1βprotein and IL-1βmRNA in LPS group were (961.00 ± 83.50) pg / m L and 5.59 ± 0.87, respectively, which were significantly higher than those in LPS group (96.33 ± 24.58 pg / m L) and 1.05 ± 0.12 (P <0.05) of control group, and microglial cells were activated. The levels of IL-1β protein and mRNA in LPS + NPY group were (411.33 ± 55.00) pg / m L and 1.93 ± 0.45, respectively, which were significantly lower than those in LPS group (P <0.05), while the level of IL-1β protein expression and mRNA expression in IBP3226 + NPY + LPS group were (886.00 ± 97.53) pg / m L and 4.51 ± 0.71, respectively, which were significantly higher than those in LPS + NPY group (P <0.05). There was no significant difference between LPS group and IBP3226 + NPY + LPS group. NPY group and control group had no statistical significance. Conclusion NPY reduces the biological activity of microglial cells and inhibits the production of IL-1β by acting on the NPY Y1 receptor.