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目的:探讨不同冷却温度和冷却时间对大鼠难治性癫癎模型海马神经细胞存活率的影响。方法:①6μmol.L-1荷包牡丹碱(Bic)+4-氨基吡啶(4-AP)联合刺激培养的大鼠海马组织切片(OHS)诱发癎样放电,制作难治性癫癎OHS模型;②在36℃和25℃培养1~24 h;③在36℃、30℃、25℃、20℃和15℃培养6 h;④分别测定OHS神经细胞的碘化丙啶(PI)摄取量。结果:Bic+4-AP联合刺激3 h后,摄取PI的神经细胞首先出现在海马的CA2区。随着刺激时间的延长,摄取PI神经细胞的分布范围扩散到其他区域,以CA3区最为明显。25℃冷却处理可以显著抑制神经细胞的PI摄取,然而,20℃、15℃深低温冷却处理可进使神经细胞的PI摄取。结论:①培养的大鼠海马组织CA2区的神经细胞对Bic+4-AP联合刺激最敏感,神经细胞的损伤与兴奋性刺激时间和海马的区域有关;②冷却处理神经细胞的保护作用与冷却的温度和时间有关。
Objective: To investigate the effects of cooling temperature and cooling time on the survival rate of hippocampal neurons in rats with intractable epilepsy. Methods: ① OHS induced by 6 μmol·L -1 biculline (Bic) + 4-aminopyridine (4-AP) was cultured in hippocampus to induce refractory epilepsy OHS model; The cells were cultured at 36 ℃ and 25 ℃ for 1 ~ 24 h. ③ The cells were cultured at 36 ℃, 30 ℃, 25 ℃, 20 ℃ and 15 ℃ for 6 h. ④ The contents of propidium iodide (PI) in OHS neurons were measured respectively. Results: After Bic + 4-AP stimulation for 3 h, PI-stimulated neurons first appeared in hippocampus CA2 region. With the extension of stimulation time, the distribution of uptake of PI nerve cells spread to other areas, the most obvious area CA3. Cooling at 25 ° C significantly inhibited PI uptake by nerve cells. However, cryogenic cooling at 20 ° C and 15 ° C led to PI uptake by nerve cells. Conclusion: ①The neurons in CA2 of hippocampus of rats are most sensitive to the combined stimulation of Bic + 4-AP, and the damage of nerve cells is related to the time of excitatory stimulation and the hippocampus. ② The cooling effect on the neuroprotective effect and cooling The temperature and time related.