Anesthesia and Brain Research Institute, Shanghai Tenth People's Hospital, School of Medicine, Tongji University, Shanghai, China; Department of Anesthesiology, Shanghai First Maternity and Infant Hospital, School of Medicine, Tongji University, Shanghai, China.
Anesthesia and Brain Research Institute, Shanghai Tenth People's Hospital, School of Medicine, Tongji University, Shanghai, China.
Br J Anaesth. 2022 Oct;129(4):544-554. doi: 10.1016/j.bja.2022.04.027. Epub 2022 Jun 11.
Environmental factors contribute to autism spectrum disorder. Fentanyl, one of the most widely used opioid analgesics in anaesthesia, can induce neurotoxicity, but its role in autism remains unknown. We determined whether fentanyl induced autism-like behaviours in young mice and the underlying mechanisms.
Young male and female mice received fentanyl at postnatal days 6, 8, and 10, and performed behavioural tests, including three-chamber social preference, elevated plus maze, grooming behaviour, and open-field test, from postnatal days 30-32. Expression of Grin2b, the gene encoding the GluN2B subunit of the N-methyl-d-aspartate receptor, was assessed in the anterior cingulate cortex of male mice using fluorescence in situ hybridisation histochemistry. We used bisulfite target sequencing to determine Grin2b hypermethylation sites after fentanyl treatment. In the specific activation and rescue experiments, we injected the mu opioid receptor agonist [D-Ala, N-MePhe, Gly-ol]-enkephalin (DAMGO) or Grin2b overexpression lentivirus into the anterior cingulate cortex of male mice.
Fentanyl induced autism-like behaviours in both young male and female mice, and downregulated Grin2b expression (0.49-fold [0.08] vs 1.00-fold [0.09]; P<0.01) and GluN2B protein amounts (0.38-fold [0.07] vs 1.00-fold [0.12]; P<0.01) in the anterior cingulate cortex through hypermethylation of Grin2b. The mu-opioid receptor antagonist naloxone and overexpression of Grin2b in anterior cingulate cortex attenuated the fentanyl-induced effects, whereas DAMGO injection into the anterior cingulate cortex induced autism-like behaviours.
These data suggest that fentanyl induces autism-like behaviours in young mice via an epigenetic mechanism. Further research is required to determine possible clinical relevance to autism risk.
环境因素促成了自闭症谱系障碍。芬太尼是麻醉中最广泛使用的阿片类镇痛药之一，可诱导神经毒性，但它在自闭症中的作用尚不清楚。我们确定芬太尼是否会在幼鼠中引起自闭症样行为及其潜在机制。
雄性和雌性幼鼠在出生后第 6、8 和 10 天接受芬太尼处理，并在出生后第 30-32 天进行行为测试，包括三箱社交偏好、高架十字迷宫、梳理行为和旷场试验。使用荧光原位杂交组织化学评估雄性小鼠前扣带回皮层中编码 N-甲基-D-天冬氨酸受体 GluN2B 亚基的 Grin2b 基因的表达。我们使用亚硫酸氢盐靶向测序来确定芬太尼处理后 Grin2b 的高甲基化位点。在特定的激活和挽救实验中，我们将μ阿片受体激动剂 [D-Ala，N-MePhe，Gly-ol]-enkephalin（DAMGO）或 Grin2b 过表达慢病毒注射到雄性小鼠的前扣带回皮层中。
芬太尼诱导雄性和雌性幼鼠出现自闭症样行为，并通过 Grin2b 的高甲基化下调前扣带回皮层中的 Grin2b 表达（0.49 倍 [0.08] 与 1.00 倍 [0.09]；P<0.01）和 GluN2B 蛋白量（0.38 倍 [0.07] 与 1.00 倍 [0.12]；P<0.01）。前扣带回皮层中的μ阿片受体拮抗剂纳洛酮和 Grin2b 的过表达减弱了芬太尼引起的作用，而 DAMGO 注射到前扣带回皮层中则诱导了自闭症样行为。
这些数据表明，芬太尼通过表观遗传机制在幼鼠中引起自闭症样行为。需要进一步研究以确定其对自闭症风险的可能临床相关性。
