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目的:探讨Nurr-1基因对小胶质细胞活化状态及其微环境的影响。方法:分离培养新生SD大鼠小胶质细胞,纯化、鉴定并分别用脂多糖(LPS)刺激活化和过表达Nurr1基因。实验随机分为小胶质细胞组、Nurr1过表达组、LPS处理组、Nurr1过表达+LPS处理组。ELISA分析过表达Nurr1基因对不同状态小胶质细胞分泌TNF-α、IL-1等炎症相关因子和BDNF、GDNF和PDNF等神经营养因子的影响。结果:(1)小胶质细胞混合培养、纯化、CD11b/c免疫细胞化学鉴定为阳性,纯度在>95%;(2)Nurr1基因过表达小胶质细胞转染阳性率>90%;(3)ELISA法检测结果显示:过表达Nurr1基因的小胶质细胞显著降低了TNF-α、IL-1的分泌,并促使了BDNF、GDNF和PDNF等神经营养因子的分泌。结论:Nurr1基因可抑制小胶质细胞的活化和减少炎症相关因子的产生,同时可促进GDNF、BDNF、PDNF等与DA能神经元存活和成熟等相关的神经营养因子的分泌。
Objective: To investigate the effect of Nurr-1 gene on microglial activation and its microenvironment. Methods: The microglia of neonatal SD rats were isolated and cultured, and purified and identified. The Nurr1 gene was activated and overexpressed by lipopolysaccharide (LPS) respectively. The experiment was randomly divided into microglial group, Nurr1 overexpression group, LPS treatment group, Nurr1 overexpression + LPS treatment group. ELISA analysis of Nurr1 gene on different state microglia secretion of TNF-α, IL-1 and other inflammatory-related factors and BDNF, GDNF and PDNF and other neurotrophic factors. Results: (1) The microglial cells were mixed culture and purified. The positive rate of CD11b / c immunocytochemistry was more than 95%. (2) The positive rate of Nurr1 overexpression microglia transfection was> 90%. 3) ELISA results showed that microglia overexpression of Nurr1 significantly decreased the secretion of TNF-α and IL-1 and promoted the secretion of neurotrophic factors such as BDNF, GDNF and PDNF. CONCLUSION: Nurr1 can inhibit the activation of microglia and reduce the production of inflammation-related factors. It can also promote the secretion of neurotrophic factors such as GDNF, BDNF and PDNF, which are related to the survival and maturation of DA neurons.