目的 研究白藜芦醇通过缺氧诱导因子-1α（HIF-1α）/磷酸果糖激酶（PFK）信号轴减少小胶质细胞糖酵解，促进大鼠脊髓损伤后的神经修复过程的作用机制。方法 选用SD大鼠构建继发性脊髓损伤模型，在模型基础上ip白藜芦醇（25、50、100 mg/kg）干预7 d，甲苯胺蓝和HE染色观察脊髓组织中尼氏小体数量和组织病变；ELISA检测脊髓组织炎性因子白细胞介素（IL）-1β、IL-6、肿瘤坏死因子-α（TNF-α）含量；免疫荧光染色观察脊髓组织HIF-1α在离子钙结合衔接分子1（IBA-1）表达和定位；蛋白质印迹检测脊髓组织M1标志物[诱导型一氧化氮合酶（iNOS）、CD86]、M2标志物[精氨酸酶（Arg1）、CD163]、HIF-1α、肌肉丙酮酸激酶同工酶（PKM2）、葡萄糖转运蛋白3（GLUT3）、己糖激酶（HK）-1和乳酸脱氢酶A（LDHA）蛋白表达。构建脂多糖（LPS）诱导小鼠小胶质细胞（BV2）炎症模型，在LPS（100 ng/mL）基础上加入白藜芦醇（30 μmol/L）和HIF-1α激动剂（DMOG，1 mmol/L）干预24 h。CCK-8检测细胞活性，免疫荧光检测细胞中CD86和CD163阳性表达；ELISA检测细胞炎性因子IL-1β、IL-6、TNF-α含量；蛋白质印迹检测HIF-1α、PKM2、GLUT3、HK-1、LDHA的蛋白表达水平。结果 与模型组比较，白藜芦醇组组织病变减轻，尼氏体数量增加，组织中IL-1β、IL-6、TNF-α、iNOS、CD86、HIF-1α、PKM2、GLUT3、HK-1、LDHA水平降低，Arg1、CD163水平升高（P＜0.05、0.01）；细胞结果显示，与模型组比较，白藜芦醇组细胞中IL-1β、IL-6、TNF-α、CD86、HIF-1α、PKM2、GLUT3、HK-1、LDHA水平降低，CD163水平升高（P＜0.01）。HIF-1α激动剂DMOG可以抑制白藜芦醇上述效果。结论 白藜芦醇通过抑制HIF-1α/PKM2介导的糖酵解代谢重编程，调控小胶质细胞M2型极化倾向，进而减轻脊髓损伤后炎症损伤。

Objective To investigate the mechanism of resveratrol promotes the nerve repair process after spinal cord injury by reducing microglia glycolysis through HIF-1α/PKM2 signaling axis.Methods Spinal cord injury model was established by using SD rats, and the rats were intraperitoneally injected with resveratrol (25, 50, and 100 mg/kg) for 7 days. Toluidine blue and HE staining were used to observe the number and pathological changes of Nissl bodies in the spinal cord tissue. ELISA was used to detect the content of IL-1β, IL-6, and TNF-α in spinal cord tissue. Immunofluorescence staining was used to observe the expression and localization of HIF-1α at IBA-1 protein. The protein expressions of M1 markers (iNOS, CD86), M2 markers (Arg1, CD163), HIF-1α, PKM2, GLUT3, HK-1, and LDHA in spinal cord tissue were detected by Western blotting. Microglia (BV2) inflammation model induced by lipopolysaccharide (LPS) was established, and resveratrol (30 mmol/L) and HIF-1α agonist (DMOG, 1 mmol/L) were added to LPS (100 ng/mL) for 24 h. CCK-8 was used to detect the cell viability, and immunofluorescence was used to detect the protein expression of CD86 and CD163 in the cells. ELISA was used to detect the levels of IL-1β, IL-6, and TNF-α. The protein expression levels of HIF-1α, PKM2, GLUT3, HK-1, and LDHA were detected by Western blotting.Results Compared with the model group, resveratrol groups alleviated the tissue lesions, increased the number of Nissl bodies, and decreased the levels of IL-1β, IL-6, TNF-α, iNOS, CD86, HIF-1α, PKM2, GLUT3, HK-1, and LDHA in the tissues, the levels of Arg-1 and CD163 were increased (P < 0.05, 0.01). Cell results showed that compared with the model group, the levels of IL-1β, IL-6, TNF-α, CD86, HIF-1α, PKM2, GLUT3, HK-1, and LDHA in the cell supernatant were decreased, but the level of CD163 was increased (P < 0.01). HIF-1α agonist DMOG could inhibit the above effects of resveratrol.Conclusions Resveratrol can regulate the M2 polarization of microglia by inhibiting HIF-1α/ PKM2-mediated glycolytic metabolic reprogramming, thereby alleviating inflammatory injury after spinal cord injury.

R285.5

延安市科技计划项目（2023-SFGG-039）