N-乙酰半胱氨酸通过介导mCalpain/TRPC6蛋白表达对癫痫大鼠学习认知功能的影响  

Effects of N-acetylcysteine on learning and cognitive function of epilepsy in rats through mediating mCalpain/TRPC6 protein expression

作　　者：罗湘蓝 刘平 苏雯靓 王苏亚 阎乃宽[1] 陈建国[1] 苏华龙[1] Luo Xianglan;Liu Ping;Su Wenliang;Wang Suya;Yan Naikuan;Ch en Jianguo;Su Hualong(Chongqing Mental Health Center,Chongqing 400000,China)

机构地区：[1]重庆市精神卫生中心,重庆400000

出　　处：《脑与神经疾病杂志》2021年第6期392-396,F0003,共6页Journal of Brain and Nervous Diseases

基　　金：重庆市精神卫生中心科研项目(2018-yjkt-05)。

摘　　要：目的探讨N-乙酰半胱氨酸对癫痫发作大鼠治疗后学习认知功能的影响及其机制。方法48只SD大鼠,随机分为对照组,癫痫组,N-乙酰半胱氨酸组(150mg·kg^(-1))和瞬时受体电位阳离子通道6(TRPC6)抑制剂SKF96365组(0.06mg·kg^(-1)),每组12只。以匹罗卡品(300mg·kg^(-1))点燃大鼠癫痫发作,造模前7d,各组给对应剂量药物。观察匹罗卡品腹腔注射后0~180 min大鼠癫痫发作情况,癫痫发作3h后,给予7.5mg·kg^(-1)地西泮缓解癫痫发作。各组大鼠再继续给药观察7d,进行水迷宫测试,检测大鼠学习认知功能。水迷宫测试结束后取脑组织用于Nissl染色和Western Blot实验。结果行为学结果显示,匹罗卡品成功点燃大鼠癫痫发作,并造成大鼠认知功能损伤,而N-乙酰半胱氨酸与SKF96365能够缓解大鼠癫痫发作并改善大鼠认知功能损伤。与对照组相比,癫痫组中钙蛋白酶(mCalpain)蛋白和TRPC6蛋白表达显著升高,Nissl染色显示海马区神经元损伤。此外,癫痫组中,磷酸化的细胞外调节蛋白激酶(p-ERK)、磷酸化的环腺苷酸应答元件结合蛋白(p-CREB)和脑源性神经营养因子(BDNF)表达水平显著降低。然而,N-乙酰半胱氨酸的预给药和治疗能够降低mCalpain蛋白表达,抑制TRPC6蛋白表达,减少神经元凋亡,并增加p-ERK、p-CREB和BDNF蛋白表达。结论匹罗卡品能够点燃大鼠癫痫症状,并造成大鼠学习认知功能损伤,N-乙酰半胱氨酸通过抑制mCalpain蛋白表达,减少TRPC6激动,改善大鼠癫痫发作,此外,ERK/CREB/BDNF通路的激动与大鼠学习认知功能改善相关。Objective To investigate the effect of N-acetylcysteine on cognitive function of rats with status epilepticus and its mechanism.Methods Forty-eight SD rats were randomly divided into control group,epilepsy group,N-Acetyl-L-cysteine(NAC)group(150mg·kg^(-1))and inhibition of transient receptor potential canonical 6 SKF96365(0.06mg·kg^(-1)),12 rats in each group.Pilocarpine(300mg·kg^(-1))was used to induce status epilepticus in rats.Seven days before the drug given,each group was administrated with the correspond drug.Status epilepticus were observed in rats from 0 to 180 minutes after injected with pilocarpine i.p.After 3 hours of status epilepticus,7.5 mg·kg^(-1)diazepam was given to relieve status epilepticus.Rats in each group continued to be administered for another 7 days,and water maze tests were performed to detect the cognitive function of the rats.After the water maze tests,brain tissue was taken for Nissl staining and Western Blot experiments.Results Behavioral results showed that pilocarpine successfully induced status epilepticus in rats and caused cognitive impairment in rats,while NAC and TRPC6 inhibitor SKF96365 could alleviate status epilepticus in rats and improved cognitive impairment.Compared with the control group,mCalpain protein and TRPC6 protein expression were significantly increased in the epilepsy group,and Nissl staining showed neuronal damage in the hippocampus.In addition,the levels of Phospho-extracellular regulated protein kinases(p-ERK),Phospho-cAMP-response element binding protein(p-CREB),and Brain-derived neurotrophic factor(BDNF)were significantly reduced in the epilepsy group.However,pre-administration and treatment of N-acetylcysteine can reduce mCalpain protein expression,inhibit TRPC6 protein expression,reduce neuronal apoptosis,and increase p-ERK,p-CREB and BDNF protein expression.Conclusion Pilocarpine can induce the status epilepticus and cause cognitive impairment in rats.NAC can reduce TRPC6 agitation and improve status epilepticus in rats by inhibiting mCalpain prote

关 键 词：癫痫 认知功能 N- 乙酰半胱氨酸 钙蛋白酶 瞬时受体电位阳离子通道6 

分 类 号：R742.1[医药卫生—神经病学与精神病学]