大脑前额皮层Nrg1基因敲低对小鼠精神分裂症样行为形成的影响  

Effects of Nrg1 gene knockdown in prefrontal cortex on formation of schizophrenia-like phenotypes in mice

作　　者：丛启杰 曾荣森 刘耘溪 孙竞[1] CONG Qijie;ZENG Rongsen;LIU Yunxi;SUN Jing(School of Pharmacy, Jiangsu University, Zhenjiang Jiangsu 212013, China)

机构地区：[1]江苏大学药学院,江苏镇江212013

出　　处：《江苏大学学报（医学版）》2022年第3期213-218,共6页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(81503051);中国博士后科学基金面上项目(2017M621488)。

摘　　要：目的:研究成年小鼠大脑前额皮层神经调节素1(neuregulin 1,Nrg1)基因敲低对精神分裂症样行为的影响及其潜在机制。方法:取成年雄性C57BL/6小鼠45只,基于旷场实验将小鼠分为3组:正常对照组,阴性对照组和Nrg1-siRNA组,每组15只。采用脑立体定位对阴性对照组和Nrg1-siRNA组小鼠前额皮层两侧部位分别注射2μL阴性对照siRNA和Nrg1-siRNA,正常对照组注射2μL PBS。采用实时荧光定量PCR和蛋白免疫印迹法分别检测小鼠前额皮层Nrg1 mRNA和NRG1蛋白表达,通过前脉冲抑制实验、旷场实验和高架十字迷宫实验评价前额皮层Nrg1基因敲低对小鼠行为的影响,采用蛋白免疫印迹法检测小鼠前额皮层囊泡γ-氨基丁酸转运体(vesicular GABA transporter,VGAT)和囊泡谷氨酸转运体1(vesicular glutamate transporter 1,VGLUT1)蛋白表达水平。结果:与正常对照组相比,Nrg1-siRNA组小鼠前额皮层Nrg1 mRNA和NRGI蛋白表达水平显著降低(P均<0.01)。行为学结果显示,与正常对照组相比,Nrg1-siRNA组小鼠前脉冲抑制率显著降低(P<0.05),自发性运动的总距离和平均速度显著增加(P均<0.01),进入开放臂的次数百分比和时间百分比显著减少(P均<0.01);蛋白免疫印迹结果显示,与正常对照组相比,Nrg1-siRNA组小鼠前额皮层VGLUT1/VGAT蛋白表达比值显著增加(P<0.01)。结论:大脑前额皮层Nrg1基因敲低可导致小鼠出现以前脉冲抑制损伤、自发活动量增加以及焦虑为特征的精神分裂症样行为,其机制可能与抑制性和兴奋性神经递质转运蛋白平衡失调有关。Objective:To explore the effect of neuregulin 1(Nrg1)knockdown in prefrontal cortex(PFC)on the formation of schizophrenia-like phenotypes in mice and its potential mechanism.Methods:A total of 45 adult male C57BL/6 mice were divided into 3 groups with 15 mice in each group based on open field test,including control group,negative control group,Nrg1-siRNA group.Stereotaxic surgery was performed to inject 2μL negative control siRNA and Nrg1-siRNA in mice PFC of negative control group and Nrg1-siRNA group,while control group were injected with 2μL PBS.Nrg1 mRNA and NRG1 protein expression in prefrontal cortex were detected by qRT-PCR and Western blotting.The mice with Nrg1 gene knockdown in PFC were evaluated on the prepulse inhibition,open field test,and elevated plus maze.Western blotting was used to detect the expression of vesicular GABA transporter(VGAT)and vesicular glutamate transporter 1(VGLUT1).Results:The expression of Nrg1 mRNA and NRG1 protein in the PFC in Nrg1-siRNA group were significantly decreased compared with control group(P<0.01).In behavior tests,compared with control group,Nrg1-siRNA group showed lower prepulse inhibition(P<0.05),higher distance and speed of spontaneous movement(both P<0.01),and the percentage of times and time to enter the open arm was significantly reduced(both P<0.01).The ratio of VGLUT1/VGAT expression in Nrg1-siRNA group was significantly increased compared with control group.Conclusion:Nrg1 gene knockdown in mice PFC could lead to significant schizophrenia-like behavior characterized by prepulse inhibition disorder,increased spontaneous movement and anxiety,which may be related to the imbalance between inhibitory and excitatory neurotransmitter transporters.

关 键 词：神经调节素1 精神分裂症 小鼠 前额皮层 脑立体定位注射 

分 类 号：R749.3[医药卫生—神经病学与精神病学]