Wang Wenda, Lin Qiujin, Liu Liru, Mai Hanhui, Tang Hongmei, Xu Kaishou
Department of Rehabilitation, Guangzhou Women and Children's Medical Center, Guangzhou Medical University, Guangzhou, Guangdong, China.
Department of Rehabilitation, Liuzhou Hospital, Guangzhou Women and Children's Medical Center, Liuzhou, Guangxi, China.
J Biochem Mol Toxicol. 2025 Sep;39(9):e70466. doi: 10.1002/jbt.70466.
Autism spectrum disorder (ASD) is a complex neurodevelopmental condition characterized by impaired social interaction, communication deficits, and repetitive behaviors. However, the underlying molecular mechanisms remain elusive. This study aims to investigate the role of cyclin-dependent kinase inhibitor 1 A (CDKN1A) in ASD. This study integrated multi-omics bioinformatics analysis to identify differentially expressed genes (DEGs) related to oxidative stress in ASD. Hub genes were screened using machine learning models. In vivo, an ASD rat model was established by maternal lipopolysaccharide (LPS) injection. Behavioral tests (open field, three-chamber social, morris water maze) were performed. Histopathology change was observed by hematoxylin-eosin staining. In vitro, LPS-stimulated BV2 microglia were treated with IL-17A for feedback experiments. Enzyme-linked immunosorbent assay was carried out to measure inflammatory factors and oxidative stress indicators. Western blot was used to detect protein expression. Bioinformatics analysis revealed 30 DEGs, with CDKN1A emerging as a prominent hub gene associated with oxidative stress. ASD model rats exhibited behavioral deficits, neuroinflammation, and hippocampal neurodegeneration. CDKN1A knockdown significantly attenuated these phenotypes, improving social interaction, reducing anxiety-like behaviors, and enhancing spatial learning and memory. Moreover, IL-17 pathway was screened as downstream pathway of CDKN1A. CDKN1A silencing suppressed LPS-induced apoptosis, inflammation, and oxidative stress in BV2 microglial cells, which was weakened by IL-17A. CDKN1A drives ASD pathogenesis via IL-17 pathway activation. Its suppression mitigates neuroinflammation, oxidative stress, and behavioral impairments, establishing CDKN1A as a novel therapeutic target for ASD. Trial Registration: Clinical trial number: Not applicable.
自闭症谱系障碍（ASD）是一种复杂的神经发育疾病，其特征为社交互动受损、沟通缺陷和重复行为。然而，其潜在的分子机制仍不清楚。本研究旨在探讨细胞周期蛋白依赖性激酶抑制剂1A（CDKN1A）在ASD中的作用。本研究整合多组学生物信息学分析，以鉴定与ASD氧化应激相关的差异表达基因（DEG）。使用机器学习模型筛选枢纽基因。在体内，通过母体注射脂多糖（LPS）建立ASD大鼠模型。进行行为测试（旷场试验、三室社交试验、莫里斯水迷宫试验）。通过苏木精-伊红染色观察组织病理学变化。在体外，用IL-17A处理LPS刺激的BV2小胶质细胞进行反馈实验。进行酶联免疫吸附测定以测量炎症因子和氧化应激指标。使用蛋白质印迹法检测蛋白质表达。生物信息学分析揭示了30个DEG，其中CDKN1A作为与氧化应激相关的突出枢纽基因出现。ASD模型大鼠表现出行为缺陷、神经炎症和海马神经退行性变。CDKN1A基因敲低显著减轻了这些表型，改善了社交互动，减少了焦虑样行为，并增强了空间学习和记忆。此外，IL-17通路被筛选为CDKN1A的下游通路。CDKN1A沉默抑制了LPS诱导的BV2小胶质细胞凋亡、炎症和氧化应激，而IL-17A减弱了这种抑制作用。CDKN1A通过激活IL-17通路驱动ASD发病机制。其抑制减轻了神经炎症、氧化应激和行为障碍，确立了CDKN1A作为ASD的新型治疗靶点。试验注册：临床试验编号：不适用。
