Department of Anesthesiology, Nanjing Drum Tower Hospital, The Affiliated Hospital Medical School, Nanjing University, No. 321 of Zhongshan Road, Nanjing, 210008, China.
BMC Med. 2024 May 7;22(1):189. doi: 10.1186/s12916-024-03413-z.
Sleep loss is a common public health problem that causes hyperalgesia, especially that after surgery, which reduces the quality of life seriously.
The 48-h sleep restriction (SR) mouse model was created using restriction chambers. In vivo imaging, transmission electron microscopy (TEM), immunofluorescence staining and Western blot were performed to detect the status of the blood-spinal cord barrier (BSCB). Paw withdrawal mechanical threshold (PWMT) was measured to track mouse pain behavior. The role of infiltrating regulatory T cells (Tregs) and endothelial cells (ECs) in mouse glycolysis and BSCB damage were analyzed using flow cytometry, Western blot, CCK-8 assay, colorimetric method and lactate administration.
The 48-h SR made mice in sleep disruption status and caused an acute damage to the BSCB, resulting in hyperalgesia and neuroinflammation in the spinal cord. In SR mice, the levels of glycolysis and glycolysis enzymes of ECs in the BSCB were found significantly decreased [CON group vs. SR group: CD31Glut1 cells: p < 0.001], which could cause dysfunction of ECs and this was confirmed in vitro. Increased numbers of infiltrating T cells [p < 0.0001] and Treg population [p < 0.05] were detected in the mouse spinal cord after 48-h SR. In the co-cultured system of ECs and Tregs in vitro, the competition of Tregs for glucose resulted in the glycolysis disorder of ECs [Glut1: p < 0.01, ENO1: p < 0.05, LDHα: p < 0.05; complete tubular structures formed: p < 0.0001; CCK8 assay: p < 0.001 on 24h, p < 0.0001 on 48h; glycolysis level: p < 0.0001]. An administration of sodium lactate partially rescued the function of ECs and relieved SR-induced hyperalgesia. Furthermore, the mTOR signaling pathway was excessively activated in ECs after SR in vivo and those under the inhibition of glycolysis or co-cultured with Tregs in vitro.
Affected by glycolysis disorders of ECs due to glucose competition with infiltrating Tregs through regulating the mTOR signaling pathway, hyperalgesia induced by 48-h SR is attributed to neuroinflammation and damages to the barriers, which can be relieved by lactate supplementation.
睡眠不足是一种常见的公共卫生问题，会导致痛觉过敏，尤其是手术后，这严重降低了生活质量。
使用限制室创建了 48 小时睡眠限制 (SR) 小鼠模型。进行体内成像、透射电子显微镜 (TEM)、免疫荧光染色和 Western blot 以检测血脊髓屏障 (BSCB) 的状态。通过测量爪撤回机械阈值 (PWMT) 来跟踪小鼠的疼痛行为。使用流式细胞术、Western blot、CCK-8 测定、比色法和乳酸给药分析浸润性调节性 T 细胞 (Tregs) 和内皮细胞 (ECs) 在小鼠糖酵解和 BSCB 损伤中的作用。
48 小时的 SR 使小鼠处于睡眠中断状态，并导致 BSCB 急性损伤，导致脊髓痛觉过敏和神经炎症。在 SR 小鼠中，BSCB 中 ECs 的糖酵解和糖酵解酶水平明显降低[CON 组与 SR 组：CD31Glut1 细胞：p <0.001]，这可能导致 ECs 功能障碍，这在体外得到了证实。在 48 小时的 SR 后，在小鼠脊髓中检测到浸润性 T 细胞数量增加[<0.0001]和 Treg 群体增加[<0.05]。在体外 ECs 和 Tregs 的共培养系统中，Tregs 对葡萄糖的竞争导致 ECs 的糖酵解紊乱[Glut1：p <0.01，ENO1：p <0.05，LDHα：p <0.05；完全形成管状结构：p <0.0001；CCK8 测定：24h 时 p <0.001，48h 时 p <0.0001；糖酵解水平：p <0.0001]。乳酸钠给药部分恢复了 ECs 的功能，并缓解了 SR 诱导的痛觉过敏。此外，体内 SR 后 ECs 中的 mTOR 信号通路过度激活，以及体外糖酵解抑制或与 Tregs 共培养的 ECs。
受浸润性 Tregs 通过调节 mTOR 信号通路与葡萄糖竞争导致的 ECs 糖酵解紊乱的影响，48 小时 SR 诱导的痛觉过敏归因于神经炎症和屏障损伤，通过补充乳酸可以缓解。
