体外激活的小胶质细胞对神经元细胞的作用研究

发布时间：2018-01-07 06:34

本文关键词：体外激活的小胶质细胞对神经元细胞的作用研究　出处：《北京协和医学院》2011年硕士论文　论文类型：学位论文

【摘要】：目的1.建立和完善大鼠脑皮质小胶质细胞原代培养的实验方法和条件；2.探索原代培养的小胶质细胞经脂多糖(LPS)体外激活后对神经元细胞的影响；3.通过丁基苯酞(NBP)对小胶质细胞激活过程的影响,探讨国家Ⅰ类新药NBP可能的新的作用靶点。方法原代培养大鼠脑皮质小胶质细胞和神经元细胞；利用LPS刺激小胶质细胞,建立原代培养的小胶质细胞体外激活的细胞模型；提取LPS激活的小胶质细胞的培养基上清液作用于神经元细胞,通过四甲基偶氮唑盐(MTT)检测神经元细胞的存活率：在加入LPS作用前用NBP处理小胶质细胞,通过观察小胶质细胞的形态,ELISA法定量检测炎性因子IL-1β、TNF-α及IL-10的释放,硝酸还原法检测炎性介质一氧化氮(NO),以及MTT检测神经元细胞经其培养基上清液作用后的存活率,综合评估NBP对小胶质细胞激活的影响。结果成功培养出数量多、纯度高的大鼠脑皮质小胶质细胞：LPS激活的小胶质细胞的培养基上清液可致神经元细胞的存活率下降；NBP处理后的小胶质细胞经LPS作用后,仍呈现为激活形态,其释放的NO减少,IL-1β、TNF-α及IL-10的含量无明显变化,经其培养基上清液作用后的神经元细胞存活率较对照组升高,且丁基苯酞的上述作用呈现一定的剂量依赖性。结论激活的小胶质细胞对神经元细胞起毒性作用；NBP不能抑制小胶质细胞的激活,但能通过抑制激活的小胶质细胞生成N0,从而减轻激活的小胶质细胞对神经元细胞的毒性作用,并具有剂量依赖性。
[Abstract]:Objective 1 . To establish and improve the experimental methods and conditions of primary culture of microglial cells in rat cerebral cortex ; 2 . To explore the effect of primary cultured microglial cells on neuronal cells following activation of lipopolysaccharide ( LPS ) in vitro ; 3 . Effects of butylphthalide ( NBP ) on the activation of microglial cells . Methods Primary cultured rat cortical microglial cells and neuronal cells were cultured in primary culture . The cells were stimulated with LPS to stimulate microglial cells . The cell survival rate of neurons was determined by MTT assay . The effects of NBP on the activation of microglial cells were assessed by means of MTT assay . The effects of NBP on the activation of microglial cells were assessed by MTT assay . Results The survival rate of the neurons was decreased after the LPS - activated microglial cells were activated by LPS . The survival rate of the neurons after NBP was decreased , the survival rate of IL - 1尾 , TNF - 伪 and IL - 10 increased , and the survival rate of the neurons after the supernatant was increased compared with the control group , and the above - mentioned effects of butylphthalide showed a dose - dependent manner . Conclusion The activated microglial cells play a toxic role in neuronal cells . NBP does not inhibit the activation of microglial cells , but can inhibit the activation of microglial cells to produce N0 , thus reducing the toxicity of activated microglial cells to neuronal cells and dose - dependent .

【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R329.2

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