神经肽Y对小神经胶质细胞活化状态和生成IL-1β的影响

The effect of NPY on the activation of microglia and IL-1βproduction

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 explore the effect of NPY on activation of primary microglia and the production of in-terleukin-1β. Methods Rat primary cortical microglia was cultured and divided into control group, LPS group, NPY＋LPS group, NPY group and BIBP3226＋NPY＋LPS group. Microglia in control group were incubated with serum-free me-dium for 6 h;microglia in LPS group were incubated with serum-free medium plus LPS for 6 h;microglia in NPY＋LPS group were incubated with serum-free medium plus NPY and LPS for 6 h; microglia cells in NPY group were incubat-ed in serum-free medium plus NPY for 6 h; microglia cells in BIBP3226＋NPY＋LPS group were incubated in se-rum-free medium including BIBP3226 、NPY and LPS for 6 h. After 6 h , Primary cultured microglia were stained us-ing IBA-1 antibody and examined under the fluorescence microscope. The protein levels of IL-1βin the culture media and the mRNA expression levels of IL-1βin the microglia of different groups were detected using the methods of Elisa and RT-PCR. Results After 6 h, the contents of IL-1 βin the culture media and the mRNA expression levels of IL-1βin the cells of LPS group increased remarkably compared with control group （P〈0.05） and the microglia were activat- ed. Compared with LPS group, the contents of IL-1 βin the culture media. the mRNA expression levels of IL-1β and the activity of microglia in LPS＋NPY group were significantly decreased .Compared with LPS＋NPY group, the contents of IL-1βin the culture media. the mRNA expression levels of IL-1β and the activity of microglia in BIBP3226＋NPY＋LPS group were increased （P〈0.05）. There were no significant differences in the contents of IL-1βin the culture media. the mRNA expression levels of IL-1βand the activity of microglia between BIBP3226＋NPY＋LPS group and LPS group or between NPY group and the control group. Conclusion NPY can inhibit the biological activity of microglia and IL-1βproduction through NPY Y1 receptorin the microglia.