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目的在PC12细胞中建立模拟永久性局部脑梗死的持续性氧糖剥夺模型。方法按照常规细胞培养方法,制备PC12细胞不同时间(5 min、10 min、15 min、30 min、45 min、1 h、3 h、6 h、12 h、24 h和48 h)持续性氧糖剥夺模型,采用MTT法检测细胞的存活率,以评估持续性氧糖剥夺对PC12细胞的损伤作用。继而进行氧、血清和葡萄糖3种因素分别交叉缺失5 min后的损伤效应研究,同样采用MTT法分析细胞的存活率。结果 PC12细胞在持续性氧糖剥夺模型条件下的半数存活时间为10.3 min,进而发现低氧和缺乏血清对细胞存活率影响不大,而培养基中葡萄糖浓度对细胞的存活率影响较大,且呈一定的浓度相关性。结论在建立持续性氧糖剥夺模型的基础上确定了PC12细胞的半数存活时间,并明确了氧、血清和葡萄糖对细胞存活率的影响,为揭示局部脑缺血低氧损伤相关疾病的机制提供了重要参考资料。
Objective To establish a continuous model of oxygen deprivation in PC12 cells that simulates permanent partial cerebral infarction. Methods According to the conventional cell culture method, the sustained oxygenation of PC12 cells was performed at different time (5 min, 10 min, 15 min, 30 min, 45 min, 1 h, 3 h, 6 h, 12 h, 24 h and 48 h) The model was deprived, the survival rate of cells was detected by MTT method to evaluate the effect of continuous oxygen glucose deprivation on PC12 cells. Then the oxygen, serum and glucose three kinds of factors were cross-missing 5 min after the injury effect, the same MTT analysis of cell viability. Results The median survival time of PC12 cells under continuous glucose deprivation model was 10.3 min, and then found that hypoxia and lack of serum had little effect on cell survival rate, while glucose concentration in culture medium had a significant effect on cell survival rate, And a certain concentration-related. Conclusion The half-survival time of PC12 cells was established based on the model of continuous oxygen-glucose deprivation, and the effect of oxygen, serum and glucose on the cell survival rate was clarified. It is provided to reveal the mechanism of hypoxic injury related to focal cerebral ischemia Important reference materials.