Celastrol (cel) was one of the earliest isolated and identified chemical constituents of Tripterygium wilfordii Hook. f. Based on a cel probe (cel-p) that maintained the bioactivity of the parent compound, the targets of cel in cerebral ischemia–reperfusion (I/R) injury were comprehensively analyzed by a quantitative chemical proteomics method. We constructed an oxygen–glucose deprivation (OGD) model in primary rat cortical neurons and a middle cerebral artery occlusion (MCAO) model in adult rats to detect the direct binding targets of cel in cerebral I/R. By combining various experimental methods, including tandem mass tag (TMT) labeling, mass spectrometry, and cellular thermal shift assay (CETSA), we revealed the targets to which cel directly bound to exert neuroprotective effects. We found that cel inhibited the proinflammatory activity of high mobility group protein 1 (HMGB1) by directly binding to it and then blocking the binding of HMGB1 to its inflammatory receptors in the microenvironment of ischemia and hypoxia. In addition, cel rescued neurons from OGD injury in vitro and decreased cerebral infarction in vivo by targeting HSP70 and NF-κB p65. Cel exhibited neuroprotective and anti-inflammatory effects by targeting HSP70 and NF-κB p65 and directly binding to HMGB1 in cerebral I/R injury.

中文翻译：

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Celastrol 通过直接结合 HMGB1 蛋白在脑缺血再灌注中发挥神经保护作用

Celastrol (cel) 是最早分离和鉴定的雷公藤化学成分之一。F。基于维持母体化合物生物活性的细胞探针（cel-p），采用定量化学蛋白质组学方法综合分析细胞在脑缺血再灌注（I/R）损伤中的作用靶点。我们在原代大鼠皮质神经元中构建了氧-葡萄糖剥夺 (OGD) 模型和在成年大鼠中构建了大脑中动脉闭塞 (MCAO) 模型，以检测 cel 在脑 I/R 中的直接结合靶点。通过结合各种实验方法，包括串联质量标签 (TMT) 标记、质谱和细胞热位移测定 (CETSA)，我们揭示了 cel 直接结合以发挥神经保护作用的靶标。我们发现 cel 通过直接与高迁移率族蛋白 1 (HMGB1) 结合，然后在缺血和缺氧的微环境中阻断 HMGB1 与其炎症受体的结合来抑制其促炎活性。此外，cel 通过靶向 HSP70 和 NF-κB p65 在体外从 OGD 损伤中拯救神经元并减少体内脑梗塞。Cel 通过靶向 HSP70 和 NF-κB p65 并在脑 I/R 损伤中直接与 HMGB1 结合，表现出神经保护和抗炎作用。