Esketamine Enhances Post-Stroke Microglial Proliferation and Attenuates Neuroinflammation by Disrupting the Src-Pannexin1 Interaction. Esketamine Enhances Post-Stroke Microglial Proliferation and Attenuates Neuroinflammation by Disrupting the Src-Pannexin1 Interaction

Previous research has suggested the potential of esketamine in alleviating post-stroke neuroinflammation, yet the underlying mechanism remained elusive. In our research, we have observed that esketamine increased the number of M2 anti-inflammatory phenotype microglia in the infarcted area after stroke, and this phenomenon is partly associated with the suppression of Panx1. Further research revealed that esketamine suppresses the phosphorylation of sarcoma kinase (Src) at the tyrosine 416 (Y416) site and inhibits the interaction between Src and Panx1, ultimately suppressing the function of Panx1. However, we found esketamine's inhibitory effect on post-stroke Panx1 activation is the underlying mechanism driving the increase in unactivated microglia, yet esketamine's influence on microglial polarization is mediated through a mechanism independent of Panx1.Our research suggests that esketamine, as a drug already used in clinical anesthesia, could potentially provide a solution during the periprocedural period to mitigate the damage caused by aseptic inflammation post-stroke. Overall design: To further explore the reasons behind the increase in Src following Panx1 inhibition, we subjected the BV2 microglial cell line to glucose-oxygen deprivation and inhibited Panx1 using 10panx1.By comparing the RNA-seq results of the group subjected solely to glucose-oxygen deprivation with those of the 10panx1+ glucose-oxygen deprivation group, we were able to identify pathways related to changes in Src.

既往研究已提示艾司氯胺酮（esketamine）在缓解脑卒中后神经炎症方面具有潜力，但其具体分子机制仍未阐明。
本研究观察到，艾司氯胺酮可增加脑卒中后梗死灶区域内M2型抗炎表型小胶质细胞的数量，该现象与潘兴蛋白1（Panx1）的功能抑制存在部分关联。
进一步机制探究显示，艾司氯胺酮可抑制肉瘤激酶（Src）在酪氨酸416（Y416）位点的磷酸化水平，并阻断Src与Panx1之间的相互作用，最终抑制Panx1的功能活性。
不过本研究发现，艾司氯胺酮对脑卒中后Panx1活化的抑制作用，是驱动未活化小胶质细胞数量增多的核心机制；但艾司氯胺酮对小胶质细胞极化的调控作用，则通过不依赖于Panx1的独立途径介导。
本研究表明，作为已应用于临床麻醉的药物，艾司氯胺酮或可在围手术期为减轻脑卒中后无菌性炎症所致的脑组织损伤提供潜在干预策略。
整体实验设计：为进一步探究Panx1抑制后Src表达上调的潜在机制，我们将BV2小胶质细胞系置于氧糖剥夺（glucose-oxygen deprivation）环境中，并采用10panx1抑制Panx1活性。通过对比仅接受氧糖剥夺处理的对照组与10panx1联合氧糖剥夺处理组的RNA测序（RNA-seq）结果，我们成功筛选出与Src表达变化相关的信号通路。