Department of Neurovascular Disease, School of Medicine, Shanghai Fourth People's Hospital, Tongji University, 1279 Sanmen Road, Shanghai, 200080, China.
Neurovascular Center, Changhai Hospital, Naval Medical University, Shanghai, China.
Neuromolecular Med. 2023 Jun;25(2):301-311. doi: 10.1007/s12017-023-08736-3. Epub 2023 Feb 7.
Stroke is a leading cause of death, with a continuously increasing incidence. As a metabolic process that catabolizes glucose pyruvate and provides adenosine triphosphate (ATP), glycolysis plays a crucial role in different diseases. Phosphoglycerate kinase 1 (PGK1) facilitates energy production with biosynthesis in many diseases, including stroke. However, the exact role of PGK1/glycolysis in stroke remains to be elucidated. A rat model of middle cerebral artery occlusion (MCAO) was used to mimic ischemia/reperfusion injuries. Oxygen glucose deprivation/re-oxygenation (OGD/R) was used to induce injury to highly aggressively proliferating immortalized (HAPI) rat microglial cells. The extracellular acidification rate (ECAR) was determined using an XFe24 Extracellular Flux Analyzer. ATP, lactate dehydrogenase, tumor necrosis factor-alpha, and interleukin-6 levels were measured using commercial kits. Chromatin immunoprecipitation assay was performed to examine the interaction between H3K27ac or p300 and the PGK1 promoter region. PGK1 was either knocked down or overexpressed by lentivirus. Thus, to examine its role in stroke, real-time polymerase chain reaction and immunoblotting were used to measure gene expression. The expression of PGK1 was increased and associated with M1 polarization and glycolysis in MCAO rat models. OGD/R promoted M1 polarization and HAPI microglial cell inflammation by regulating glycolysis. Silencing PGK1 reduced OGD/R-increased M1 polarization, inflammation, and glycolysis. Conversely, the overexpression of PGK1 promoted HAPI microglial cell inflammation by regulating glycolysis. The mechanism showed that histone acetyltransferase p300 promoted PGK1 expression through H3K27 acetylation. Finally, data indicated that silencing PGK1 inhibited microglia M1 polarization, inflammation, and glycolysis in MCAO rat models. PGK1 could promote ischemia/reperfusion injury-induced microglial M1 polarization and inflammation by regulating glycolysis, which might provide a novel direction in developing new therapeutic medications for preventing or treating stroke.
中风是导致死亡的主要原因之一，其发病率呈持续上升趋势。糖酵解作为一种代谢过程，可分解葡萄糖丙酮酸并提供三磷酸腺苷（ATP），在许多疾病中都具有重要作用。磷酸甘油酸激酶 1（PGK1）在包括中风在内的许多疾病中促进生物合成和能量产生。然而，PGK1/糖酵解在中风中的确切作用仍有待阐明。使用大脑中动脉闭塞（MCAO）大鼠模型模拟缺血/再灌注损伤。使用氧葡萄糖剥夺/再氧合（OGD/R）诱导高度侵袭性增殖的永生（HAPI）大鼠小胶质细胞损伤。使用 XFe24 细胞外通量分析仪测定细胞外酸化率（ECAR）。使用商业试剂盒测定 ATP、乳酸脱氢酶、肿瘤坏死因子-α和白细胞介素-6 水平。通过染色质免疫沉淀测定法检查 H3K27ac 或 p300 与 PGK1 启动子区域之间的相互作用。通过慢病毒敲低或过表达 PGK1。因此，为了研究其在中风中的作用，使用实时聚合酶链反应和免疫印迹法测量基因表达。PGK1 的表达增加，并与 MCAO 大鼠模型中的 M1 极化和糖酵解相关。OGD/R 通过调节糖酵解促进 M1 极化和 HAPI 小胶质细胞炎症。沉默 PGK1 可减少 OGD/R 增加的 M1 极化、炎症和糖酵解。相反，PGK1 的过表达通过调节糖酵解促进 HAPI 小胶质细胞炎症。机制表明组蛋白乙酰转移酶 p300 通过 H3K27 乙酰化促进 PGK1 表达。最后，数据表明，沉默 PGK1 可抑制 MCAO 大鼠模型中小胶质细胞 M1 极化、炎症和糖酵解。PGK1 可以通过调节糖酵解促进缺血/再灌注损伤诱导的小胶质细胞 M1 极化和炎症，这可能为开发预防或治疗中风的新治疗药物提供新的方向。
