自闭症谱系障碍(ASD)是一种严重的神经发育障碍。疼痛感觉异常是ASD常见的临床症状,严重影响ASD患者及其家属的生活质量。然而,潜在机制尚不清楚.它被认为与神经元的兴奋性和离子通道的表达有关。在这里,我们证实,在BTBRT+Itpr3tf/J(BTBR)ASD小鼠模型中,基线疼痛和完全弗氏佐剂(CFA)诱导的慢性炎性疼痛受损。背根神经节(DRG)的RNA测序(RNA-seq)分析,这与ASD模型小鼠的疼痛密切相关,提示KCNJ10(编码Kir4.1)的高表达可能是ASD痛觉异常的重要因素。通过蛋白质印迹进一步验证了Kir4.1的水平,RT-qPCR,和免疫荧光。通过抑制Kir4.1,BTBR小鼠的疼痛不敏感性得到改善,证实Kir4.1的高表达水平与ASD疼痛敏感性降低高度相关。同时,我们发现,CFA诱发炎性疼痛后,焦虑行为和社会新颖性认知发生了改变.抑制Kir4.1后,BTBR小鼠的刻板行为和社会新颖性识别也得到了改善。Further,我们发现谷氨酸转运体的表达水平,兴奋性氨基酸转运蛋白1(EAAT1),和兴奋性氨基酸转运蛋白2(EAAT2)在BTBR小鼠的DRG中增加,但在抑制Kir4.1后降低。这表明Kir4.1可能通过调节谷氨酸转运蛋白在改善ASD的疼痛不敏感中起关键作用。总之,我们的发现揭示了Kir4.1在ASD疼痛不敏感中的可能机制和作用,利用生物信息学分析和动物实验,为ASD的临床针对性干预提供理论依据。
Autism spectrum disorder (ASD) is a severe neurodevelopmental disorder. Abnormal pain sensation is a common clinical symptom of ASD that seriously affects the quality of life of patients with ASD and their families. However, the underlying mechanism is unclear. It is believed to be related to the excitability of neurons and the expression of ion channels. Herein, we confirmed that baseline pain and Complete Freund\'s adjuvant (CFA)-induced chronic inflammatory pain were impaired in the BTBR T+ Itpr3tf/J (BTBR) mouse model of ASD. RNA sequencing (RNA-seq) analyses of the dorsal root ganglia (DRG), which are closely related to pain in ASD model mice, revealed that high expression of KCNJ10 (encoding Kir4.1) might be an important factor in ASD pain sensation abnormalities. The levels of
Kir4.1 were further verified by western blotting, RT-qPCR, and immunofluorescence. By inhibiting
Kir4.1, the pain insensitivity of BTBR mice improved, confirming that a high expression level of
Kir4.1 was highly correlated with decreased pain sensitivity in ASD. Meanwhile, we found that the anxiety behaviours and the social novelty recognition were changed after CFA induced inflammatory pain. And after inhibiting
Kir4.1, the stereotyped behaviours and social novelty recognition of BTBR mice were also improved. Further, we found that the expression levels of glutamate transporters, excitatory amino acid transporter 1 (EAAT1), and excitatory amino acid transporter 2 (EAAT2) were increased in the DRG of BTBR mice but decreased after inhibiting Kir4.1. This suggests that Kir4.1 may play a key role in the improvement of pain insensitivity in ASD by regulating glutamate transporters. In conclusion, our findings revealed the possible mechanism and role of
Kir4.1 in the pain insensitivity in ASD, using bioinformatics analyses and animal experiments, and provided a theoretical basis for clinically targeted intervention in ASD.