中文摘要：

目的探讨钙调蛋白依赖性激酶Ⅱ（CaMKⅡ）对异氟烷神经毒性的作用及相关机制。方法 0.96 mM异氟烷处理培养1 w后的神经细胞6 h,异氟烷处理之前两天在培养基中加入CaMKⅡ抑制剂十四烷酰肉豆蔻酰-钙调蛋白自变性抑制肽（myrAIP）或激动剂钙调素（CaM）。抑制剂实验将细胞随机分为对照组、CaMKⅡ抑制剂处理组、异氟烷处理组、异氟烷加抑制剂组（n=10）,激动剂实验分为对照组、异氟烷处理组、异氟烷加CaMKⅡ激动剂组（n=10）。检测细胞活力MTT,对细胞进行Hoechst33258荧光染色观察凋亡细胞并计数。结果抑制剂实验中,与对照组相比,各处理组MTT值明显降低（P〈0.05）,凋亡神经元明显增多（P〈0.05）,异氟烷加抑制剂组的凋亡细胞明显多于单纯异氟烷处理组（P〈0.05）;激动剂实验中,异氟烷加激动剂组部分逆转了单纯异氟烷处理引起的MTT下降（P〈0.05）并减轻了单纯异氟烷处理引起的凋亡细胞增多（P〈0.05）。结论 0.96mM的异氟烷处理6h对小鼠的神经元产生了明显的毒性;CaMKⅡ抑制剂能明显加重异氟烷的神经毒性。

英文摘要：

Objective To explore the effect of calmodulin dependent kinase Ⅱ（CaMKⅡ） on isoflurane neurotoxicity and its mechanism.Methods Cerebral neurons separated from fetal mice were cultured for seven days before they were subjected to an isoflurane（0.96 mM） exploration for 6 h.CaMKII inhibitor myristoylated autocamtide-2-related inhibitory peptide（myrAIP） or its agonist calmodulin（CaM） was added to the culture medium 2 d prior to the experiment.Cells were randomly divided into control group,myrAIP group,isoflurane group,isoflurane＋myrAIP group in inhibitor experiment and control group,isoflurane group,isoflurane＋CaM group in agonist experiment（n=10）.The effects of anesthetics on cell viability and injury were assayed by methylthiazolyltetra-zolium（MTT）.Apoptotic cells were detected by Hoechst 33258 staining.Results In inhibitor experiment,MTT levels were decreased and the number of apoptotic cells was increased by different treatments compared with control group（P0.05）.The number of apoptotic cells in isoflurane＋myrAIP group was significantly lower than that of isoflurane group（P0.05）.In agonist experiment,MTT levels were increased and the number of apoptotic cells was decreased in isoflurane＋CaM group compared with isoflurane group（P0.05）.Conclusion Fetal mice neurons exposure to 0.96 mM isoflurane for 6 h can result in obvious neurotoxicity;CaMKⅡ inhibitor can significantly increase the neurotoxicity of isoflurane.