Liu Yong-Qi, Chen Gao, Wang Ke-Wei, Yan Xin-Jiang, Zhan Cheng-Peng, Yu Guo-Feng
The Second Clinical Medical School, Zhejiang Chinese Medical University, Hangzhou, Zhejiang, China.
School of Medicine, Quzhou College of Technology, Quzhou, Zhejiang, China.
J Mol Histol. 2024 Dec 30;56(1):61. doi: 10.1007/s10735-024-10336-9.
Traumatic brain injury (TBI) is a common neurosurgical emergency. As a macrophage in brain, microglia involves in secondary TBI injury. UCF-101, an Omi/HtrA2 inhibitor, protects against neurological disorders. This study aims to investigate the effects of UCF-101 in TBI and its mechanism. Mouse microglia cell BV2 cells were exposed to 1 µg/mL LPS to construct TBI in vitro models. Following CCK8 assay, cells were treated with LPS + UCF-101 (2, 5, 10 µM), LPS + Compound C (AMPK inhibitor, 20 µM), and LPS + UCF-101 + Compound C groups. With lactate dehydrogenase (LDH) content detection, ELISA and qRT-PCR assays were used to measure proinflammatory factors. Biomarkers of M1 (CD16/32 and iNOS) and M2 phenotypes (CD206), as well as AMPK/NF-κB pathway-related protein expression were assessed by flow cytometry, immunofluorescence, and Western blot methods. There was a decrease in M1 phenotype biomarkers and an increase in M2 phenotype biomarkers after UCF-101 treatment. UCF-101 exposure reduced TNF-α, LDH, IL-1β, IL-6, IL-8, p-NF-κB p65/NF-κB p65, and activated p-AMPK α (T172)/AMPK α (T172) expression. Importantly, further Compound C treatment counteracted these effects of UCF-101. In conclusion, UCF-101 ameliorates TBI by promoting microglia M2 polarization via AMPK/NF-κB pathways in LPS-induced BV2 cells, providing solid scientific foundation for clinical application of UCF-101 in TBI treatment.
创伤性脑损伤（TBI）是一种常见的神经外科急症。作为脑内的巨噬细胞，小胶质细胞参与了创伤性脑损伤的继发性损伤。Omi/HtrA2抑制剂UCF-101可预防神经疾病。本研究旨在探讨UCF-101对创伤性脑损伤的影响及其机制。将小鼠小胶质细胞BV2细胞暴露于1μg/mL脂多糖（LPS）以构建体外创伤性脑损伤模型。经CCK8检测后，将细胞分为LPS+UCF-101（2、5、10μM）、LPS+Compound C（AMPK抑制剂，20μM）以及LPS+UCF-101+Compound C组进行处理。通过检测乳酸脱氢酶（LDH）含量，采用酶联免疫吸附测定（ELISA）和实时定量聚合酶链反应（qRT-PCR）检测促炎因子。通过流式细胞术、免疫荧光和蛋白质免疫印迹法评估M1（CD16/32和诱导型一氧化氮合酶（iNOS））和M2表型（CD206）的生物标志物，以及AMPK/核因子κB（NF-κB）信号通路相关蛋白的表达。UCF-101处理后，M1表型生物标志物减少，M2表型生物标志物增加。UCF-101可降低肿瘤坏死因子-α（TNF-α）、LDH、白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）、白细胞介素-8（IL-8）、磷酸化核因子κB p65（p-NF-κB p65）/核因子κB p65的表达，并激活磷酸化腺苷酸活化蛋白激酶α（p-AMPK α（T172））/腺苷酸活化蛋白激酶α（T172）的表达。重要的是，进一步给予Compound C处理可抵消UCF-101的这些作用。总之，UCF-101通过在脂多糖诱导的BV2细胞中经由AMPK/NF-κB信号通路促进小胶质细胞M2极化来改善创伤性脑损伤，为UCF-101在创伤性脑损伤治疗中的临床应用提供了坚实的科学依据。
