419 Synergistic White Matter Injury Secondary to Chronic Cerebral Hypoperfusion and Air Pollution is Reduced by Microglia-Specific TLR4-Knockdown

Abstract

INTRODUCTION: Air pollution is recognized as a neurotoxin which damages white matter and increases the risk of cognitive decline. Our previous work in a murine model found that air pollution in the setting of chronic cerebral hypoperfusion (CCH) leads to a synergistic deleterious effect on white matter. Air pollution studies have shown that increased microglial activation was associated with neurotoxicity. We hypothesize that inhibition of a microglial-specific TLR4 pathway will decrease the neurodegenerative effects that normally result from combined CCH and air pollution. METHODS: Our air pollution model used eight weeks of standardized Diesel Exhaust Particles (DEP) from National Institute of Science and Technology (DEP NIST2975). We performed bilateral carotid stenosis (BCAS) surgery to induce CCH using Wild Type (WT) and TLR4 flx/flx (CX3CR1CreER +/-) (TLR4-KD) in 2-month-old male mice. WT and TLR4-KD mice were randomized into eight groups as follows: WT-filtered air (FA), WT-DEP, WT-FA-BCAS, WT-DEP-BCAS, TLR4-KD-FA, TLR4-KD-DEP, TLR4-KD-FA-BCAS, and TLR4-KD-DEP-BCAS. Neurotoxicity was quantified with immunohistochemistry of degraded myelin basic protein (dMBP), ionized calcium-binding adaptor molecule-1 (Iba-1), and complement (C5a). RESULTS: DEP exposure resulted in increased white matter injury in corpus callosum (dMBP), worsened by the presence of CCH (p = 0.004) and significantly attenuated by TLR4-KD (p = 0.005). DEP exposure resulted in increased white matter microglial activation (Iba-1), worsened by the presence of CCH (p < 0.0001) and significantly attenuated by TLR4-KD (p = 0.04).DEP exposure resulted in increased white matter complement activation (C5a), worsened by the presence of CCH (p < 0.0001) and significantly attenuated by TLR4-KD (p < 0.0001). CONCLUSIONS: Microglial-specific TLR4-KD reduces air pollution-induced neurotoxicity measured by white matter injury, microglial activation, and neuroinflammation.