奥拉西坦调节AKT/GSK-3β信号通路对孤独症小鼠的改善作用

Improvement effect of olaxetan on autistic mice by regulating the AKT/GSK-3βsignaling pathway

目的探讨奥拉西坦(ORC)调节蛋白激酶B(AKT)/糖原合成酶激酶-3β(GSK-3β)信号通路对孤独症小鼠的改善作用。方法通过注射丙戊酸(VPA)建立孤独症孕鼠,取其产下的雄性幼鼠作为研究对象,将造模成功的幼鼠随机分为VPA组、ORC-L组、ORC-H组、ORC-H+LY294002组,每组10只,并以注射生理盐水的正常孕鼠产下的雄性幼鼠为空白组,随后评估社交交互与偏好能力;埋珠实验检测重复刻板行为;新物体实验评估学习与记忆能力;H-E染色观察海马组织病理损伤;试剂盒检测前额叶组织中氧化应激指标——超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、丙二醛(MDA)水平。Westernblot检测AKT/GSK-3β信号通路相关蛋白及脑源性神经营养因子(BDNF)蛋白表达。结果与空白组相比,VPA组小鼠社会交往指数、社会偏好指数、识别指数、SOD、CAT水平、BDNF、p-GSK-3β/GSK-3β、p-AKT/AKT表达明显降低,埋珠数量、MDA水平明显增加(P<0.05);与VPA组相比,ORC-L组、ORC-H组的上述指标发生逆转,组间有差异(P<0.05);LY294002逆转了ORC-H对孤独症小鼠的改善作用。结论ORC调节AKT/GSK-3β信号通路改善孤独症小鼠症状。

Objective To explore the improvement effect of oxiracetam(ORC)on autism mice by regulating the proteinkinase B(AKT)/glycogen synthase kinase-3β(GSK-3β)signaling pathway.Methods Autistic pregnant mice were established by injecting valproic acid(VPA),and male offspring were selected as the study subjects.The successfully modeled offspring were randomly grouped into VPA group,ORC-L group,ORC-H group,ORC-H+LY294002 group,with 10 mice in each group.Male offspring from normal pregnant mice injected with physiological saline were used as the blank group.Social interaction and preference abilities were evaluated.The marbles bury experiment was applied to detect repetitive stereotyped behavior.New object experiment was applied to evaluate learning and memory abilities.H-E staining was applied to observe pathological damage in hippocampal tissue.Reagent kits were applied to detect oxidative stress indicators such as superoxide dismutase(SOD),catalase(CAT),and malondialdehyde(MDA)levels in frontal lobe tissue.Western blot was applied to detect the expression of AKT/GSK-3βsignaling pathway related proteins and brain derived neurotrophic factor(BDNF)protein.Results Compared with the blank group,the social interaction index,social preference index,recognition index,SOD,CAT levels,BDNF,p-GSK-3β/GSK-3β,and p-AKT/AKT expression of mice in the VPA group were greatly reduced,the number of buried marbles and MDA level were greatly increased(P<0.05).Compared with VPA group,the above indexes in ORC-L group and ORC-H group were reversed,and there were differences among groups(P<0.05).LY294002 reversed the improving effects of ORC-H on autistic mice.Conclusion ORC improves symptoms in autistic mice by regulating the AKT/GSK-3βsignaling pathway.