Fan Linlin, Zeng Xin, Jiang Yutong, Zheng Danyang, Wang Han, Qin Qian, Li Mengyue, Wang Hui, Liu Hao, Liang Shengjun, Pang Xiuming, Shi Shanyi, Wu Lijie, Liang Shuang
Department of Child and Adolescent Health, Public Health College, Harbin Medical University, Harbin 150081, China.
Outpatient Department, The Second Affiliated Hospital of Heilongjiang University of Chinese Medicine, Harbin 150001, China.
Phytomedicine. 2024 Jun;128:155386. doi: 10.1016/j.phymed.2024.155386. Epub 2024 Feb 1.
Maternal immune activation (MIA) is a significant factor inducing to autism spectrum disorder (ASD) in offspring. The fundamental principle underlying MIA is that inflammation during pregnancy impedes fetal brain development and triggers behavioural alterations in offspring. The intricate pathogenesis of ASD renders drug treatment effects unsatisfactory. Traditional Chinese medicine has strong potential due to its multiple therapeutic targets. Yigansan, composed of seven herbs, is one of the few that has been proven to be effective in treating neuro-psychiatric disorders among numerous traditional Chinese medicine compounds, but its therapeutic effect on ASD remains unknown.
Yigansan improves MIA-induced ASD-like behaviours in offspring by regulating the IL-17 signalling pathway.
Pregnant C57BL/6J mice were intraperitoneally injected with poly(I:C) to construct MIA models and offspring ASD models. Network analysis identified that the IL-17A/TRAF6/MMP9 pathway is a crucial pathway, and molecular docking confirmed the binding affinity between the monomer of Yigansan and target proteins. qRT-PCR and Western blot were used to detect the expression levels of inflammatory factors and pathway proteins, immunofluorescence was used to detect the distribution of IL-17A, and behavioural tests were used to evaluate the ASD-like behaviours of offspring.
We demonstrated that Yigansan can effectively alleviate MIA-induced neuroinflammation of adult offspring by regulating the IL-17A/TRAF6/MMP9 pathway, and the expression of IL-17A was reduced in the prefrontal cortex. Importantly, ASD-like behaviours have been significantly improved. Moreover, we identified that quercetin is the effective monomer for Yigansan to exert therapeutic effects.
Overall, this study was firstly to corroborate the positive therapeutic effect of Yigansan in the treatment of ASD. We elucidated the relevant molecular mechanism and regulatory pathway involved, determined the optimal therapeutic dose and effective monomer, providing new solutions for the challenges of drug therapy for ASD.
母体免疫激活（MIA）是导致后代患自闭症谱系障碍（ASD）的重要因素。MIA的基本原理是孕期炎症会阻碍胎儿大脑发育并引发后代行为改变。ASD复杂的发病机制使得药物治疗效果不尽人意。中药因其具有多个治疗靶点而具有强大潜力。由七种草药组成的益肝散是众多中药复方中少数已被证明对神经精神疾病有效的方剂之一，但其对ASD的治疗效果尚不清楚。
益肝散通过调节白细胞介素-17（IL-17）信号通路改善MIA诱导的后代ASD样行为。
对怀孕的C57BL/6J小鼠腹腔注射聚肌胞苷酸（poly(I:C)）以构建MIA模型和后代ASD模型。网络分析确定IL-17A/肿瘤坏死因子受体相关因子6（TRAF6）/基质金属蛋白酶9（MMP9）通路是关键通路，分子对接证实益肝散单体与靶蛋白之间的结合亲和力。采用实时定量聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法检测炎症因子和通路蛋白的表达水平，免疫荧光法检测IL-17A的分布，行为测试评估后代的ASD样行为。
我们证明益肝散可通过调节IL-17A/TRAF6/MMP9通路有效减轻成年后代MIA诱导的神经炎症，前额叶皮质中IL-17A的表达降低。重要的是，ASD样行为得到了显著改善。此外，我们确定槲皮素是益肝散发挥治疗作用的有效单体。
总体而言，本研究首次证实了益肝散治疗ASD的积极疗效。我们阐明了相关分子机制和调控通路，确定了最佳治疗剂量和有效单体，为ASD药物治疗面临的挑战提供了新的解决方案。
