【摘要】：目的：隨著電力事業(yè)的發(fā)展,人群接觸極低頻磁場的時間和強度愈來愈大,人們對極低頻磁場的效應(yīng)也愈加關(guān)注。早期的研究表明極低頻磁場可以誘導(dǎo)細胞分子水平的改變,如：細胞縫隙連接功能(GJIC)下降、細胞膜表皮生長因子受體(EGFR)和腫瘤壞死因子受體(TNFR)聚簇,以及基因和蛋白質(zhì)表達譜發(fā)生改變等等。細胞自噬是機體重要的一種防御和保護機制,它大量降解并重復(fù)利用長壽命蛋白質(zhì)、大分子物質(zhì)以及細胞內(nèi)受損的細胞器等,使細胞能夠在饑餓環(huán)境下得以生存,細胞自噬和神經(jīng)退行性疾病、衰老以及腫瘤相關(guān)。有研究發(fā)現(xiàn)電離輻射如γ射線,非電離輻射如紫外線等均會誘導(dǎo)細胞自噬效應(yīng),本研究初步探索與人們?nèi)粘Ｉ蠲芮邢嚓P(guān)的工頻磁場(50Hz0.4mT)對中國倉鼠肺成纖維細胞(Chinese hamster lung cells, CHL)的細胞自噬的誘導(dǎo)。方法：CHL細胞培養(yǎng)24小時,更換新鮮的培養(yǎng)基后進行50Hz0.4mT工頻磁場暴露30分鐘、2小時、6小時、12小時或者24小時,磁場暴露后通過以下三種方法檢測細胞自噬水平：透射電鏡觀察細胞內(nèi)空泡變化,激光共聚焦顯微鏡檢測細胞內(nèi)GFP-LC3定位自噬小體情況以及Western Blot實驗檢測細胞內(nèi)LC3-Ⅱ表達。結(jié)果：透射電子顯微鏡觀察到,工頻磁場暴露30分鐘和24小時誘導(dǎo)細胞內(nèi)空泡增加；激光共聚焦顯微鏡觀察到,在GFP-LC3高表達的CHL細胞內(nèi),工頻磁場暴露不同時間點誘導(dǎo)GFP-LC3亮點標(biāo)記的自噬小體增加(P0.05),30分鐘和24小時升高趨勢較為明顯；Western Blot技術(shù)檢測發(fā)現(xiàn)磁場暴露促進自噬的特異性標(biāo)記蛋白LC3-Ⅱ表達增加(P0.05),30分鐘和24小時升高趨勢較為明顯；加入自噬小體與溶酶體融合的阻斷劑氯喹后,磁場暴露30 min自噬流量增加(P0.05)。結(jié)論：這些研究提示本實驗條件下50Hz0.4mT工頻磁場暴露可能誘導(dǎo)CHL細胞自噬效應(yīng)。
[Abstract]:Objective: with the development of electric power industry, the time and intensity of people contact with very low frequency magnetic field are increasing, and people pay more attention to the effect of very low frequency magnetic field. Early studies have shown that very low frequency magnetic fields can induce cell molecular changes, such as cell gap junction function (GJIC) decreased, cell membrane epidermal growth factor receptor (EGFR) and tumor necrosis factor receptor (TNFR) cluster, And gene and protein expression profile changes and so on. Autophagy is an important defense and protection mechanism. It degrades and reuses long-lived proteins, macromolecules and damaged organelles in order to make cells survive in hungry environment. Autophagy and neurodegenerative diseases, aging, and tumours are associated. Some studies have found that ionizing radiation such as 緯-rays and non-ionizing radiation such as ultraviolet radiation can induce autophagy effect. The purpose of this study was to explore the induction of autophagy of Chinese hamster lung fibroblasts (Chinese hamster lung cells, CHL) by power frequency magnetic field (50Hz0.4mT), which is closely related to people's daily life. Methods the cells were cultured for 24 hours. The fresh culture medium was replaced with 50Hz0.4mT power frequency magnetic field exposure for 30 minutes, 2 hours, 6 hours, 12 hours, or 24 hours, respectively. The level of autophagy was detected by three methods after magnetic field exposure: the changes of intracellular vacuoles were observed by transmission electron microscope (TEM), the intracellular GFP-LC3 localization autophagy was detected by laser confocal microscopy, and the expression of LC3- 鈪，

本文編號：2193288