系统性抑制Toll样受体4改善母体免疫激活子代大鼠的ASD样行为及其神经炎症

Systemic inhibition of TLR4 improves autism spectrum disorder-like behaviors and neuroinflammation in offspring rats following maternal immune activation

摘要

目的 探讨Toll样受体4(TLR4)的特异性阻断剂TAK-242对母体免疫激活(MIA)子代大鼠孤独症谱系障碍(ASD)样行为及神经炎症的影响,为ASD早期干预提供实验依据。方法 将30只SD孕鼠随机分为对照组、LPS模型组、TAK-242干预组每组各10只。于妊娠第9.5天(G9.5),LPS组和TAK-242组注射100μg/kg LPS,对照组注射等量PBS,TAK-242组注射LPS前2h予以TAK-242 3mg/kg腹腔注射。取雄性仔鼠于生后6~8周龄进行行为学实验评估其社会行为与认知功能;采用ELISA法检测孕鼠和仔鼠血清IL-6、TNF-α水平,qRT-PCR和免疫组化分别检测仔鼠前额叶皮质炎症因子表达及小胶质细胞形态与标志物Iba-1表达情况。结果 与对照组相比,LPS组孕鼠及仔鼠血清IL-6、TNF-α显著升高,差异均有统计学意义(P<0.001)。行为学显示,LPS组仔鼠出现埋珠数增多、中央区停留时间缩短、社交指数下降及学习记忆能力受损(F=9.811,14.490,31.750,5.205,P<0.05),前额叶皮质促炎因子(IL-1β、IL-6、NF-κB p65、iNOS)mRNA表达升高,抗炎因子(IL-4、IL-10、Arg-1)mRNA表达下降(P<0.001),小胶质细胞呈活化形态(胞体增大、分支缩短),Iba-1表达升高(P<0.001)。TAK-242干预后,上述异常均得到显著改善(P<0.005)。结论 TAK-242通过抑制TLR4/NF-κB通路,改善MIA诱发的子代ASD样行为,其机制可能与调节小胶质细胞极化及减轻神经炎症有关。

Abstract

Objective To investigate the effects of TAK-242, a specific Toll-like receptor 4 (TLR4) inhibitor, on autism spectrum disorder(ASD)-like behaviors and neuroinflammation in offspring rats induced by maternal immune activation (MIA), so as to provide an experimental basis for early ASD intervention. Methods Thirty pregnant SD rats were randomly divided into control group, LPS, and TAK-242 intervention group (n=10 per group).On gestational day 9.5 (G9.5), the LPS and TAK-242 groups received an intraperitoneal injection of 100μg/kg LPS, while the control group received an equivalent volume of PBS.The TAK-242 group received a pre-injection of 3mg/kg TAK-242 2 hours before LPS administration.Male offspring underwent behavioral tests at 6 - 8 weeks of age to assess their social behavior and cognitive function.Serum levels of IL-6 and TNF-α in the dams and offspring were measured using ELISA.In the offspring, the expression of inflammatory factors in the prefrontal cortex (PFC) was detected by qRT-PCR, while microglial morphology and the expression of the marker Iba-1 were examined by immunohistochemistry. Results Compared with the control group, the LPS group showed significantly elevated serum levels of IL-6 and TNF-α in both dams and offspring (P<0.001).Offspring in the LPS group exhibited ASD-like behavioral abnormalities, including increased marble burying, reduced central zone duration in the open field, decreased social index, and impaired learning and memory (F=9.811, 14.490, 31.750, 5.250, P<0.05).Furthermore, the LPS group demonstrated up-regulated mRNA expression of pro-inflammatory factors and down-regulated mRNA expression of anti-inflammatory factors in the PFC (P<0.001), along with microglial activation (characterized by enlarged cell bodies and shortened processes) and increased Iba-1 expression.TAK-242 intervention significantly ameliorated all these abnormalities (P<0.05). Conclusions TAK-242 ameliorates MIA-induced ASD-like behaviours in offspring by inhibiting the TLR4/NF-κB signalling pathway, potentially through modulating microglial polarization and attenuating neuroinflammation.

关键词

孤独症谱系障碍 / TAK-242 / 小胶质细胞 / Toll样受体4 / 脂多糖

Key words

autism spectrum disorder / TAK-242 / microglia / Toll-like receptor 4 / lipopolysaccharide

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基金资助

新疆维吾尔自治区儿童重大疾病大数据平台与一体化防诊治体系建立(2024B03038-2)