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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肿瘤坏死因子-α(TNF-α)的影响。方法培养原代大鼠皮质小胶质细胞,经脂多糖(LPS)和NPY处理后行免疫细胞化学荧光染色,显微镜下观察LPS和NPY对小胶质细胞形态学的影响。将小胶质细胞分为对照组、LPS组、NPY+LPS组、NPY组和BIBP3226+NPY+LPS组,每组3个样本,培养各组细胞6 h。ELISA法检测培养液中TNF-α蛋白含量,RT-PCR方法检测小胶质细胞中TNF-αmRNA表达水平。结果 LPS处理后小胶质细胞处于活化状态,NPY处理后的小胶质细胞活化水平降低。LPS组和IBP 3226+NPY+LPS组培养液中TNF-α蛋白含量及细胞中TNF-αmRNA表达水平显著高于对照组(P均<0.05);LPS+NPY组TNF-α蛋白含量和TNF-αmRNA表达水平明显低于LPS组和IBP3226+NPY+LPS组(P均<0.05)。结论 NPY能够降低小神经胶质细胞的生物活性。NPY可能通过激活NPY Y1受体抑制小神经胶质细胞生成TNF-α。
Objective To investigate the effect of neuropeptide Y (NPY) on the primary microglia biological activity and the production of tumor necrosis factor-α (TNF-α). Methods Primary rat cortical microglial cells were cultured and treated with lipopolysaccharide (LPS) and NPY for immunocytochemical staining. The morphological changes of microglia were observed under microscope. The microglia were 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 h. The content of TNF-α in culture medium was detected by ELISA, and the expression of TNF-αmRNA in microglia was detected by RT-PCR. Results After LPS treatment, microglial cells were activated and the activation of microglia after NPY treatment was decreased. The levels of TNF-αmRNA and TNF-α mRNA in LPS group and IBP 3226 + NPY + LPS group were significantly higher than those in control group (all P <0.05). The levels of TNF-αprotein and TNF- αmRNA expression levels were significantly lower than the LPS group and IBP3226 + NPY + LPS group (P all <0.05). Conclusion NPY can reduce the biological activity of microglia. NPY may inhibit the production of TNF-α by microglia by activating NPY Y1 receptor.