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LUXI WANG, Mei Chen, Heping Xu; Control microglial activation by targeting the glucose transporter 1. Invest. Ophthalmol. Vis. Sci. 2016;57(12):No Pagination Specified. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
Microglial cells are critically involved in various retinal inflammatory and degenerative diseases. Immune cell activation is known to be regulated by metabolic pathways within the cells. The aim of this study was to understand how microglial activation is regulated by glucose metabolic pathway.
Primary microglial cells were cultured from 4-6 week old C57BC/6J mice. The phenotype of the cells was confirmed by CD11b/IBA-1 expression. The cells were further activated by lipopolysaccharide (100ng/ml) or interleukin 4 (20 ng/ml) with or without a GLUT1 inhibitor STF31, and the mRNA expression levels of GLUTs and inflammatory cytokines including TNFa, iNOS, IL6, CCL2, IL10 and Arginase-1 in microglial cells were examined by real-time RT-PCR. Microglial phagocytosis was conducted by using the pHrodo Green Zymosan Bioparticles Kit. Glycolysis rates were assessed by the XF analyser. Glucose uptake was determined by the accumulation of intracellular 2-deoxyglucose -6-phosphate (2-DG6P).
Microglial cells express a variety of GLUTs. Among all glucose transporters expressed by microglia GLUT1 showed the highest level of expression. The expression of GLUT1 was significantly increased after LPS but not IL-4 stimulation. Inhibition of GLUT1 with SFT31 significantly reduced the expression of TNFa, iNOS, IL6, and CCL2 in LPS treated cells. Interestingly, IL-4 induced upregulation of IL10 and Arginase1 was also significantly reduced by STF31 treatment. Furthermore, SFT31 suppressed the phagocytic activity of microglial cells. In line with the functional changes, microglial cells treated with SFT31 showed reduced glucose uptake and decreased glycolysis rate. STF31 did not affect the expression of GLUT1 at both mRNA and protein levels. The inhibitor also showed no adverse effect on the viability of microglial cells.
GLUT1 plays an important role in glucose uptake and metabolism in microglial cells. Inhibiting glucose uptake by targeting GLUT1 could suppress microglial activation. Targeting the GLUT1 may be a novel approach to manipulation microglial function and control inflammation.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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