Mechanistic exploration of miR-384 targeting PFKFB3 to regulate glycolysis and neuroinflammation in micro-glia
HAN Shi-hao
LIU Xin-yu
WANG Bin
Abstract：Objective To analyze the molecular mechanism by which miR-384 regulates inflammation in micro-glia.Methods BV2 microglial cells were treated with 100 ng/mL LPS for 24 hours to establish an in vitro model of neu-roinflammation.miR-384 mimics and mimic NC were transfected into BV2 cells,and RT-qPCR was used to detect the mRNA expression levels of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3(PFKFB3),hexokinase 2(HK2),interleukin-1β(IL-1β),and interleukin-6(IL-6).Potential targets of miR-384 were predicted using the miRDB database,and dual-luciferase reporter assays verified the targeting relationship between miR-384 and PFKFB3.The expression levels of HK2 and PFKFB3 proteins were detected by Western blot after overexpression of miR-384.Results LPS induced the expression of inflammatory genes IL-6 and IL-1β in BV2 cells and increased lactic acid levels in the cell supernatant.Overexpression of miR-384 inhibited the LPS-induced increase in lactic acid.Addi-tionally,miR-384 overexpression suppressed the upregulation of the glycolytic gene PFKFB3 induced by LPS,while inhi-bition of miR-384 further increased PFKFB3 expression.No significant effects were observed on the glycolytic gene HK2.Dual-luciferase reporter assays confirmed that PFKFB3 is a downstream target of miR-384.Furthermore,overex-pression of miR-384 inhibited the expression of IL-1β and IL-6,while inhibition of miR-384 produced the opposite effects.Conclusion miR-384 directly targets the glycolytic gene PFKFB3 and regulates LPS-induced glycolysis and neuroinflammatory responses.
Keywords：neuroinflammationmiR-384fructose-2-kinase 6-phosphate/fructose-26-diphosphatase 3glycolysis
Publication Date：2024-10-28
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：8( 1247-1254 )