Title Page

Contents

ABSTRACT ENGLISH 7

INTRODUCTION 9

MATERIAL AND METHODS 12

1. Ethics statement and care of experiment animal 12

2. Seizure induction 12

3. Tamoxifen and lactate administration 13

4. Brain Sample Preparation 14

5. Detection of live neuron 14

6. Immunofluorescence analysis 15

7. Detection of blood brain barrier disruption 17

8. Detection of zinc accumulation and lysosome 17

9. Western blot 18

10. Data analysis 19

RESULTS 20

1. Effects of amlexanox on seizure behavior and body weight change after pilocarpine-induced seizure 20

2. Amlexanox reduces PDE4B expression and pro-inflammation factor, but increases zinc in lysosomes after the seizure 20

3. Amlexanox increases autophagy and lysosomal function, but decreases neuro-inflammation 22

4. Amlexanox decreases neuronal death after pilocarpine-induced seizure 23

5. Amlexanox decreases glia cell activation after seizure 24

6. Amlexanox reduces oxidative stress and microtubule disruption after seizure 25

7. Amlexanox reduces Blood–Brain Barrier (BBB) disruption after seizure 27

DISCUSSION 28

REFERENCES 47

ABSTRACT KOREAN 53

Figure 1. (A) The experimental schedule involved the administration of pilocarpine and the assessment of seizure grades every 5 minutes until reaching... 32

Figure 2. (A) Phosphodiesterase 4B (PDE4B) staining (red) was performed 12 hours post-seizure and in the sham group. Neuronal population was visualized using... 34

Figure 3. (A-C) Western blot analysis assessed the levels of lysosome-associated membrane protein 2 (LAMP2) and light chain 3B (LC3B), markers of lysosomal... 36

Figure 4. (A) Neuronal survival was assessed using live neuron staining with Neuronal nuclear protein (NeuN) in the subiculum, CA1, cornu ammonis 3 (CA3),... 38

Figure 5. (A-C) Glial fibrillary acidic protein (GFAP, green) staining highlighted astrocytes in the CA1 region after seizure and in the sham group. Complement 3... 40

Figure 6. (A-B) Microtubules were visualized using microtubule associated protein 2 (MAP2) staining in the CA1 region, represented by green fluorescence.... 42

Figure 7. (A) The micrographs display the immunostaining of the hippocampus using anti-rat IgG, indicating blood-brain barrier (BBB) disruption. The scale bar... 44

Figure 8. Our hypothesis for the present study. A. The schematic diagram depicts the pilocarpine-induced seizure process, emphasizing the crucial steps and... 45