星形胶质细胞活化与表皮生长因子受体表达的相关性研究

发布时间：2018-07-05 12:33

本文选题：星形胶质细胞活化 + 胶原纤维酸性蛋白　；参考：《南华大学》2011年硕士论文

【摘要】：目的研究活化星形胶质细胞（Astrocyte,Ast）表皮生长因子受体（epithelialgrowth factor receptor,EGFR）的表达变化，探讨EGFR对星形胶质细胞活化所起的关键作用。方法采用振荡培养法结合差速贴壁法分离纯化培养星形胶质细胞，将得到的第二代星形胶质细胞分为对照组、活化组、抑制组：将常规用含10%FBS的DMEM培养基培养24h后的星形胶质细胞作为对照组细胞，用加入20ng/ml睫状神经营养因子、10%FBS的DMEM培养基培养24h后的星形胶质细胞为活化组细胞，将含30μmol/L Genistein、10%FBS的DMEM培养基作用于活化组细胞再培养24h后的细胞作为抑制组细胞。通过免疫荧光化学观察各组细胞的形态变化；应用半定量RT-PCR方法分析各组细胞间胶原纤维酸性蛋白(glial fibrillary acidic protein,GFAP) mRNA及EGFR mRNA表达变化。结果活化组与对照组比较，星形胶质细胞细胞数量增多，胞体变大，突起增多变长，交织成网状；RT-PCR示不仅GFAP mRNA表达增高，EGFR mMRA表达亦明显增高，与对照组比较有显著差异（P＜0.01）；应用EGFR抑制剂Genistein干预后，与活化组相比，，星形胶质细胞数量不再增多，细胞胞体变小，突起减少缩短，星形胶质细胞活化被抑制，GFAP mRNA表达下降，与活化组比较有显著差异（P＜0.01）。结论：1.活化后的星形胶质细胞EGFR表达明显上调； 2.Genistein通过抑制EGFR表达，从而可使星形胶质细胞活化受到抑制。
[Abstract]:Objective to investigate the expression of epidermal growth factor receptor (epithelialgrowth factor receptor) in activated astrocytes and to explore the key role of EGFR in the activation of astrocytes. Methods astrocytes were isolated and purified by oscillatory culture combined with differential adhesion method. The second generation astrocytes were divided into control group and activated group. In the inhibition group, astrocytes were cultured in DMEM medium containing 10s for 24 hours as control cells, astrocytes cultured in 10 S DMEM medium for 24 hours as activation group, and ciliary neurotrophic factor added in DMEM medium as control group, while astrocytes cultured in 10 S DMEM medium for 24 hours were used as control cells. The DMEM medium containing 30 渭 mol / L Genistein, 10s was used as the inhibitory group cells after 24 hours of reculture. The morphological changes of the cells were observed by immunofluorescence, and the expression of collagen fibrillary acidic protein (glial fibrillary acidic) mRNA and EGFR mRNA were analyzed by semi-quantitative RT-PCR. Results compared with the control group, the number of astrocytes increased, the body of astrocytes became larger, the processes increased and longer, and the expression of EGFR mMRA was significantly increased in the activated group compared with the control group. The expression of EGFR mMRA was not only increased in the activated group, but also increased in the interlaced reticular-shaped RT-PCR. Compared with the control group, the number of astrocytes was no longer increased, the cell body became smaller and the process decreased after the intervention of EGFR inhibitor Genistein. The expression of GFAP mRNA in astrocytes was significantly lower than that in activated group (P < 0.01). Conclusion 1. (2) Genistein inhibited the activation of astrocytes by inhibiting EGFR expression.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R329

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