本文選題：星形膠質(zhì)細(xì)胞活化 + 膠原纖維酸性蛋白　； 參考：《南華大學(xué)》2011年碩士論文

【摘要】：目的研究活化星形膠質(zhì)細(xì)胞（Astrocyte,Ast）表皮生長因子受體（epithelialgrowth factor receptor,EGFR）的表達(dá)變化，探討EGFR對(duì)星形膠質(zhì)細(xì)胞活化所起的關(guān)鍵作用。 方法采用振蕩培養(yǎng)法結(jié)合差速貼壁法分離純化培養(yǎng)星形膠質(zhì)細(xì)胞，將得到的第二代星形膠質(zhì)細(xì)胞分為對(duì)照組、活化組、抑制組：將常規(guī)用含10%FBS的DMEM培養(yǎng)基培養(yǎng)24h后的星形膠質(zhì)細(xì)胞作為對(duì)照組細(xì)胞，用加入20ng/ml睫狀神經(jīng)營養(yǎng)因子、10%FBS的DMEM培養(yǎng)基培養(yǎng)24h后的星形膠質(zhì)細(xì)胞為活化組細(xì)胞，將含30μmol/L Genistein、10%FBS的DMEM培養(yǎng)基作用于活化組細(xì)胞再培養(yǎng)24h后的細(xì)胞作為抑制組細(xì)胞。通過免疫熒光化學(xué)觀察各組細(xì)胞的形態(tài)變化；應(yīng)用半定量RT-PCR方法分析各組細(xì)胞間膠原纖維酸性蛋白(glial fibrillary acidic protein,GFAP) mRNA及EGFR mRNA表達(dá)變化。 結(jié)果活化組與對(duì)照組比較，星形膠質(zhì)細(xì)胞細(xì)胞數(shù)量增多，胞體變大，突起增多變長，交織成網(wǎng)狀；RT-PCR示不僅GFAP mRNA表達(dá)增高，EGFR mMRA表達(dá)亦明顯增高，與對(duì)照組比較有顯著差異（P＜0.01）；應(yīng)用EGFR抑制劑Genistein干預(yù)后，與活化組相比，，星形膠質(zhì)細(xì)胞數(shù)量不再增多，細(xì)胞胞體變小，突起減少縮短，星形膠質(zhì)細(xì)胞活化被抑制，GFAP mRNA表達(dá)下降，與活化組比較有顯著差異（P＜0.01）。 結(jié)論：1.活化后的星形膠質(zhì)細(xì)胞EGFR表達(dá)明顯上調(diào)； 2.Genistein通過抑制EGFR表達(dá)，從而可使星形膠質(zhì)細(xì)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.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R329

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