The effect of PFKFB3 on M1-type polarization of microglia and neuronal damage in stroke

Objective: This study aims to reveal the role of PFKFB3 (6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3) in the M1-type polarization of microglia in strokeMethods: Primary microglial cells and hippocampal neurons from mice were used. They were divided into three groups: sh-NC, sh-NC + OGD/R, and sh-PFKFB3 + OGD/R. After transfection of microglial cells for 72 hours, they were co-cultured with neurons and subjected to OGD/R. After 24 hours, the indicators were detected. Western blot was used to detect the expression of PFKFB3 protein (Western blot); flow cytometry was used to detect the proportion of CD16/32+Iba1+ M1-type microglia and the apoptosis rate of neurons.Results: Western blot results showed that compared with the sh-NC group, the expression of PFKFB3 in the cells of the sh-NC + OGD/R group was significantly increased. sh-PFKFB3 could effectively inhibit the increase of PFKFB3 caused by OGD/R (Figure 2-1A). After co-culturing microglial cells with neurons for 72 hours and subjected to OGD/R, using flow cytometry to detect the proportion of CD16/32+M1-type microglia in Iba1+ cells and the apoptosis rate of neurons, it was shown that compared with the sh-NC group, the proportion of CD16/32+M1-type microglia in the sh-NC + OGD/R group increased significantly (Figure 2-1B), and the apoptosis rate of neurons in the co-culture system also increased significantly (Figure 2-1C); while using sh-PFKFB3 to reduce the expression of PFKFB3 and then giving OGD/R treatment, all the above indicators were significantly reversed, indicating that the increase of PFKFB3 is related to the polarization of microglia in stroke, and inhibiting PFKFB3 can inhibit the M1-type polarization of microglia and reduce neuronal damage.Conclusion: Inhibiting PFKFB3 can inhibit the M1-type polarization of microglia and reduce neuronal damage Funding: National Key Clinical Specialty Construction Project Research Special Project (2409052,241007), Teaching Reform and Innovation Project of Shanxi Medical University (XJ2025019), Key Country Science and Technology Cooperation Project of Shanxi Province (202204041101007), Key Research and Development Project for Introducing High-level Scientific and Technological Talents of Lvliang City (2023RC08,2024RC19), Shanxi Province Overseas Students' Innovation and Entrepreneurship Start-up Support Program Project (SXLC-RY-2022), Shanxi Province Leading Talent Project in Science and Technology Innovation (SJYC2024009), Research Project of Shanxi Provincial Health Commission (2025YGYL010) This abstract was presented at the American Physiology Summit 2026 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.