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目的探讨诱导型一氧化氮合酶(iNOS)表达上调是否参与脂多糖(LPS)诱导的多巴胺(DA)能神经元变性。方法脑立体定位注射5μgLPS至大鼠脑黑质,观察不同时间点(6、12h,1、3、7d),iNOSmRNA及蛋白表达的动态变化;化学比色法检测黑质NO释放量以及iNOS活性改变。结果iNOSmRNA和蛋白的动态变化为6h开始出现增高,1d达高峰,3d开始下降,7d后基本消失。1d时iNOS阳性细胞数(45.30±4.63)显著高于PBS对照侧iNOS阳性细胞数(0.11±0.04)(P<0.01)、RT-PCR、Western印迹法显示1d组iNOSmRNA和iNOS蛋白表达明显多于正常对照组和PBS对照侧;同时发现iNOSmRNA和蛋白过度表达,并促进NO释放,iNOS活性升高。结论iNOS上调可能是LPS诱导DA能神经元变性机制中重要的因素之一。
Objective To investigate whether upregulation of inducible nitric oxide synthase (iNOS) is involved in the degeneration of dopaminergic neurons induced by lipopolysaccharide (LPS). Methods The stereotactic injection of 5 μg LPS into rat brain substantia nigra was used to observe the dynamic changes of iNOS mRNA and protein expression at different time points (6, 12 h, 1, 3 and 7 d). The NO release and iNOS activity change. Results The dynamic changes of iNOS mRNA and protein began to increase at 6h, peaked at 1d, decreased at 3d, and disappeared after 7d. The number of iNOS positive cells (45.30 ± 4.63) at 1d was significantly higher than that of PBS control (0.11 ± 0.04) (P <0.01). The expression of iNOS mRNA and iNOS protein in 1d group was significantly higher than that in 1d group Normal control group and PBS control side; also found that iNOS mRNA and protein overexpression, and promote NO release, iNOS activity increased. Conclusion Upregulation of iNOS may be one of the important factors in the mechanism of LPS-induced degeneration of DA neurons.