Kan Yufei, Wang Hong, Lin Huaying, Li Yongfa, Pei Shuaijie, Cui Yan, Xie Keliang, Chen Hongguang, Yu Yonghao
Department of Anesthesiology, Tianjin Institute of Anesthesiology, Tianjin Medical University General Hospital, Tianjin, PR China.
Department of Anesthesiology, Shanxi Cancer Hospital, Shanxi Hospital Affiliated to Cancer Hospital, Chinese Academy of Medical Sciences, Taiyuan, Shanxi Province, PR China.
J Neurochem. 2025 Feb;169(2):e70003. doi: 10.1111/jnc.70003.
Sepsis-associated encephalopathy (SAE) is a brain dysfunction for which no effective therapy currently exists. Recent studies suggest that transferring mitochondria from astrocytes to neurons may benefit SAE patients, though the underlying mechanism remains unclear. We cultured astrocytes and neurons from mice in vitro. Astrocytes were stimulated with lipopolysaccharide (LPS) for 24 h, and the astrocyte-conditioned medium (ACM) was collected. Neuronal cultures were then treated with ACM or mitochondria-depleted ACM (mdACM) for further analysis. Mitochondrial transfer was examined under a fluorescence microscope. Western blotting analyzed the protein expression of genes related to apoptosis and mitochondrial metabolism. RNA sequencing and mass spectrometry were employed to investigate the mechanisms underlying mitochondrial transfer. Astrocyte-derived mitochondria migrated toward and connected with LPS-exposed neurons. The addition of ACM significantly attenuated LPS-induced alterations in the proteins linked to apoptosis and mitochondrial dynamics. RNA sequencing revealed notable alterations in the transcript profile of neurons upon ACM treatment, highlighting the involvement of mitochondria metabolism, inflammation, and apoptosis-related factors. Additionally, mitochondrial transfer modified the lipid composition of neurons, increasing phosphatidylserine levels, which correlated with neuroinflammation and enriched pathways related to cytokine and MAPK signaling. Our findings suggest that astrocyte-neuron mitochondrial transfer holds therapeutic potential for alleviating SAE, possibly through the anti-inflammatory effects of lipids, particularly phosphatidylserine.
脓毒症相关脑病（SAE）是一种目前尚无有效治疗方法的脑功能障碍。最近的研究表明，将星形胶质细胞的线粒体转移到神经元可能对SAE患者有益，但其潜在机制尚不清楚。我们在体外培养了小鼠的星形胶质细胞和神经元。用脂多糖（LPS）刺激星形胶质细胞24小时，然后收集星形胶质细胞条件培养基（ACM）。然后用ACM或线粒体耗尽的ACM（mdACM）处理神经元培养物以进行进一步分析。在荧光显微镜下检查线粒体转移情况。蛋白质印迹法分析与细胞凋亡和线粒体代谢相关基因的蛋白表达。采用RNA测序和质谱分析来研究线粒体转移的潜在机制。星形胶质细胞衍生的线粒体向暴露于LPS的神经元迁移并与之连接。添加ACM可显著减轻LPS诱导的与细胞凋亡和线粒体动力学相关蛋白质的改变。RNA测序显示，ACM处理后神经元的转录谱有显著变化，突出了线粒体代谢、炎症和细胞凋亡相关因子的参与。此外，线粒体转移改变了神经元的脂质组成，增加了磷脂酰丝氨酸水平，这与神经炎症以及与细胞因子和丝裂原活化蛋白激酶信号相关的富集途径有关。我们的研究结果表明，星形胶质细胞-神经元线粒体转移可能通过脂质尤其是磷脂酰丝氨酸的抗炎作用，对缓解SAE具有治疗潜力。
