摘要
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 influence of Nurr1 in activation and the microenvironment for microglia.Methods：The microglia of neonatal SD rats was isolated and cultured.After purification and identification,the Nurr1 activation and overexpression in microglia was utilized by the Lipopolysaccharide （LPS） stimulation.The microglia was randomly divided into microglia group,Nurr1 overexpression group,LPS group and Nurr1 overexpression ＋ LPS group.Elisa was applied to detect the influence of Nurr1 overexpressed for BDNF,GDNF,PDNF,TNF-α,IL-1 and other inflam mation-associated cytokines which were secreted by the multi-microglia.Results：（1） The microglia mixed cultivation,purification and the CD1 1 b/c identification were positive,and purity was above 95%;（2）The transfection positive rate of Nurr1 overexpression in microglia was more than 90%;（3） The ELISA detection showed that the Nurr1 overexpression in microglia decreased distinctly the secretion of TNF-α and IL-1 and increased the secretion of BDNF,GDNF,PDNF.Conclusion：Nurr1 can inhibit the activation of microglia and the secretion of TNF-α and IL-1,and promote the secretion of BDNF,GDNF,PDNFneurotrophic factor which was related with dopaminergic neuronal survival and mature.
出处
《神经解剖学杂志》
CAS
CSCD
北大核心
2017年第2期160-166,共7页
Chinese Journal of Neuroanatomy
基金
国家自然科学基金(81241126
81360197)
云南省教育厅科学研究基金项目(2013C227)
云南省科技厅-昆明医学院联合专项(2014FB041)
云南省干细胞与再生医学重点实验室开放课题(701160109)
