The oral mulit-target kinase inhibitor X is inhibits oncogenic receptor tyrosine kinase (RTK) and is an effective targeted therapy for patients with metastatic colorectal cancer as well as advanced gastrointestinal stromal tumors. However, whether X can affects neuroinflammation and AD pathology is not well studied. Thus, we tested the effects of X on neuroinflammatory responses and found that X regulates LPS-induced neuroinflammatory responses in glial cells, wild-type mice, and 5x FAD mice (a mouse model of AD). X affects TLR4/AKT/STAT3 signaling to alter LPS-mediated proinflammatory cytokine levels, suggesting that X can suppress LPS-mediated neuroinflammation in vitro and in vivo. We then examined whether X can alter AD pathology and found that X -injected 5x FAD mice increased dendritic spine density, decreased A plaque levels by modulating APP processing and autophagy ATG12 levels. Importantly, X -injected 5x FAD mice significantly reduced tau phosphorylation at AT100 by downregulating tau kinase p-GSK3. Taken together, our results indicate that X can regulates AD pathology, neuroinflammation, and dendritic spine formation.
Table Of Contents
Ⅰ. INTRODUCTION 1 Ⅱ. MATERIALS AND METHODS 3 2.1 Cell lines and culture conditions 3 2.2 MTT assays 4 2.3 Reverse transcription-polymerase chain reaction (RT-PCR) 4 2.4 Real-time PCR(Q-PCR) 5 2.5 Antibodies and inhibitors 5 2.6 Western blotting 6 2.7 Immunocytochemistry 7 2.8 Cytosolic and nuclear fractionation 7 2.9 Mouse primary microglial cell and astrocyte culture 8 Ⅲ. RESULTS 9 3.1 Regorafenib regulates LPS-induced proinflammatory cytokine levels 9 3.2 Regorafenib modulates AKT/STAT3 signaling to alter neuroinflammatory responses 10 3.3 Regorafenib alters LPS-induced proinflammatory cytokine levels in primary microglia and astrocytes 15 3.4 Regorafenib attenuated LPS-induced glia activation and proinflammatory response in vivo 19 3.5 Regorafenib significantly increased dendritic spine density in primary hippocampal neurons and 5x FAD mice brain 23 3.6 Regorafenib significantly downregulated Aβ plaque levels in 5x FAD mice 25 3.7 Regorafenib significantly reduced tau phosphorylation at T212 and S214 and tau kinase GSK3β activity in 5x FAD mice brain 30 Ⅳ. DISCUSSION 35 Ⅴ. REFERENCES 40