c-Jun氨基末端激酶在孤独症谱系障碍发病机制中的作用

Role of c-Jun N-terminal kinase in the pathogenesis of autism spectrum disorder

目的检测孤独症谱系障碍(ASD)患儿的c-Jun氨基末端激酶(JNK)通路蛋白表达情况,探讨JNK在ASD发病机制中的作用。方法选择2016年6月至2017年6月于湖南省儿童医院确诊的30例ASD患儿作为ASD组,另选择30例健康儿童作为对照组。检测外周血单核淋巴细胞(PBMC)中总JNK(T-JNK)及磷酸化JNK(p-JNK)表达水平;采用儿童期孤独症评定量表(CARS)评定疾病严重程度,并对JNK活化水平与CARS评分进行相关性分析。利用基因重组腺体病毒载体转染原代神经元细胞,根据干预措施不同分为JNK过表达组和对照组,检测两组中总JNK、磷酸化的JNK/ATF-2/c-Jun、囊泡谷氨酸转运体(VGLUT)和囊泡型GABA转运体(VGAT)表达水平。结果 ASD组和对照组PBMC中T-JNK表达水平差异无统计学意义(P> 0. 05),但ASD组p-JNK表达水平以及p-JNK/T-JNK比值显著高于对照组(P <0. 05)。采用单因素回归分析评估外周血p-JNK/T-JNK比值与CARS评分相关性,结果显示JNK活化程度与疾病严重程度不具有相关性(r=0. 03,P> 0. 05)。腺病毒载体转染原代神经元细胞后,JNK过表达组中T-JNK、p-NK/ATF-2/c-Jun表达水平均显著高于对照组(P <0. 05);JNK过表达组中VGAT的表达水平均显著低于对照组(P <0. 05),VGLUT的表达水平与对照组比无明显差异(P> 0. 05)。结论 JNK信号通路可能参与了ASD的发病,但与疾病严重程度无关。上调JNK表达水平能使转录因子ATF-2和c-Jun活化,下调突触囊泡转运体中VGAT表达。

Objective To investigate the expression of c-Jun N-terminal kinase( JNK) proteins in children with autism spectrum disorder( ASD) and the role of JNK in the pathogenesis of ASD. Methods A total of 30 children with ASD who were diagnosed in Hunan Children's Hospital from June 2016 to June 2017 were enrolled as ASD group,and 30 healthy children were enrolled as control group.Western blot was used to measure the expression levels of total JNK( T-JNK) and phosphorylated JNK( p-JNK) in peripheral blood mononuclear lymphocytes( PBMCs),Childhood Autism Rating Scale( CARS) was used to evaluate disease severity,and a correlation analysis was performed for JNK activation level and CARS score. Primary neuronal cells were transfected with recombinant adenovirus vector and were divided into JNK over-expression group and control group according to the intervention measures. The expression levels of T-JNK,p-JNK,phosphorylated ATF-2( p-ATF-2),phosphorylated c-Jun( p-c-Jun),vesicular glutamate transporter( VGLUT),and vesicle-type GABA transporter( VGAT) were measured for both groups. Results There was no significant difference in the expression level of T-JNK in PBMCs between the ASD group and the control group( P 0. 05),but the ASD group had significantly higher expression level of p-JNK and p-JNK/T-JNK ratio than the control group( P 0. 05). The univariate logistic regression analysis was used to investigate the correlation between peripheral blood p-JNK/T-JNK ratio and CARS score,and the results showed that there was no correlation between JNK activation level and disease severity( r = 0. 03,P 0. 05). After the primary neuron cells were transfected with adenovirus vector,the JNK over-expression group had significantly higher expression levels of T-JNK,p-JNK,p-c-Jun,and p-ATF-2 and a significantly lower expression level of VGAT than the control group( P 0. 05),while there was no significant difference in the expression level of VGLUT between the two groups( P 0. 05). Conclusions The JNK signaling pathway may be involved in the pathogenesis of ASD,but it is not associated with disease severity. Upregulation of JNK expression levels can activate the transcription factors ATF-2 and c-Jun and downregulate the expression of VGAT in synaptic vesicle transporters.