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目的和方法 :应用胎鼠皮层细胞原代培养 ,建立神经元的体外“缺血 /再灌注”模型 ,观察神经元缺血 /再灌注后DNA链的损伤。应用PANT和TUNEL染色分别检测缺血 /再灌注后DNA单链和双链损伤。结果 :神经元缺糖缺氧 2h引起极少量细胞死亡 ,4h引起少于 30 %的细胞死亡 ,而 6～ 8h的缺糖缺氧引起的细胞死亡数量达到80 %以上 ,6h缺糖缺氧再灌注 10～ 18h ,细胞死亡达高峰 ,而在 8h缺糖缺氧再灌注 2h细胞死亡已经达高峰。在缺糖缺氧 2 ,4 ,6 ,8h再灌注 5min ,PANT阳性细胞分别达 30 % ,5 0 % ,80 % ,90 %。而在同样的情况下 ,TUNEL染色阳性细胞数没有明显增加。结论 :体外神经元缺糖缺氧再灌注早期即出现DNA链的损伤 ,且以单链损伤为主。
PURPOSE AND METHODS: Primary cultured rat cortical cells were used to establish neuronal in vitro model of “ischemia / reperfusion” and to observe the DNA damage after neuronal ischemia / reperfusion. PANT and TUNEL staining were used to detect single and double strand DNA damage after ischemia / reperfusion. Results: A very small amount of cell death was induced by hypoxia and hypoxia in neurons for 2 hours, less than 30% cell death occurred after 4 hours, while the number of cell death caused by hypoxia and hypoxia for 6 ~ 8 hours reached more than 80% Perfused with 10 ~ 18h, cell death reached the peak, and in 8h after hypoxia and hypoxia reperfusion 2h cell death reached its peak. PANT positive cells were 30%, 50%, 80% and 90% respectively at 2, 4, 6 and 8 hours after reperfusion for 5 minutes. In the same situation, the number of TUNEL-positive cells did not increase significantly. CONCLUSION: DNA strand damage occurs in the early phase of hypoxia-ischemia reperfusion in vitro neurons.