China Medical University School of Forensic Medicine, No.77 Puhe Road, Shenyang, China.
Liaoning Province Key Laboratory of Forensic Bio-evidence Sciences, Shenyang, China.
Mol Neurobiol. 2024 Apr;61(4):2049-2062. doi: 10.1007/s12035-023-03669-1. Epub 2023 Oct 16.
Ketamine as a glutamate receptor antagonist has a rapid, potent, and long-lasting antidepressant effect, but its specific mechanism is still not fully understood. Depression is associated with elevated levels of glutamate and astrocyte loss in the brain; the exploration of the relationships between ketamine's antidepressant effect and astrocytes has drawn great attention. Astrocytes and aquaporin 4 (AQP4) are essential components of the glymphatic system, which is a brain-wide perivascular pathway to help transport nutrients to the parenchyma and remove metabolic wastes. In this study, we investigated pyroptosis-associated protein Nlrp3/Caspase-1/Gsdmd-N expression in the hippocampus of mice and the toxic effect of high levels of glutamate on primary astrocytes. On this basis, the protective mechanism of ketamine is explored. A single administration of ketamine (10 mg/kg) remarkably relieved anxious and depressive behaviors in the sucrose preference test, elevated plus maze test, and forced swim test. Meanwhile, ketamine reduced the level of hippocampus Nlrp3 and the expression of its downstream molecules in chronic unpredictable mild stress (CUMS) mice model by western blot and reduced the colocalization of Gfap and Gsdmd by nearly 25% via immunofluorescent staining. Ketamine also increased the Gfap-positive cells and AQP4 expression in the hippocampus of the CUMS mice. More important, ketamine increased the distribution of the fluorescent tracer of CUMS mice. Treatment with 128 mM glutamate in cortical and hippocampus astrocytes increased the level of Nlrp3, and Gsdmd-N, and ketamine alleviated high glutamate-induced pyroptosis-associated proteins. In summary, these results suggest that high glutamate-induced astrocyte pyroptosis through the Nlrp3/Caspase-1/Gsdmd-N pathway which was inhibited by ketamine and ketamine can improve the damaged glymphatic function of the CUMS mice. The present study indicates that inhibiting astrocyte pyroptosis and promoting the glymphatic circulation function are a new mechanism of ketamine's antidepressant effect, and astrocyte pyroptosis may be a new target for other antidepressant medicines.
氯胺酮作为一种谷氨酸受体拮抗剂,具有快速、强效和持久的抗抑郁作用,但它的确切机制仍不完全清楚。抑郁症与大脑中谷氨酸水平升高和星形胶质细胞缺失有关;氯胺酮的抗抑郁作用与星形胶质细胞之间的关系的探索引起了极大的关注。星形胶质细胞和水通道蛋白 4(AQP4)是胶状淋巴系统的重要组成部分,胶状淋巴系统是一个遍布大脑的血管周围途径,有助于将营养物质输送到实质并清除代谢废物。在这项研究中,我们研究了小脑中与细胞焦亡相关的蛋白 NLRP3/Caspase-1/GSDMD-N 的表达以及高水平谷氨酸对原代星形胶质细胞的毒性作用。在此基础上,探索了氯胺酮的保护机制。单次给予氯胺酮(10mg/kg)可显著缓解蔗糖偏好测试、高架十字迷宫测试和强迫游泳测试中小鼠的焦虑和抑郁行为。同时,氯胺酮通过 Western blot 降低慢性不可预测轻度应激(CUMS)小鼠模型中海马 NLRP3 及其下游分子的表达,并通过免疫荧光染色使 Gfap 和 Gsdmd 的共定位减少近 25%。氯胺酮还增加了 CUMS 小鼠海马中的 Gfap 阳性细胞和 AQP4 表达。更重要的是,氯胺酮增加了 CUMS 小鼠荧光示踪剂的分布。皮质和海马星形胶质细胞中 128mM 谷氨酸处理增加了 NLRP3 和 Gsdmd-N 的水平,而氯胺酮缓解了高谷氨酸诱导的细胞焦亡相关蛋白。综上所述,这些结果表明,高谷氨酸通过 Nlrp3/Caspase-1/Gsdmd-N 通路诱导星形胶质细胞焦亡,而氯胺酮抑制了这种通路,氯胺酮可以改善 CUMS 小鼠受损的胶状淋巴功能。本研究表明,抑制星形胶质细胞焦亡和促进胶状淋巴循环功能是氯胺酮抗抑郁作用的新机制,星形胶质细胞焦亡可能是其他抗抑郁药物的新靶点。