Title Page
ABSTRACT
국문 초록
Contents
CHAPTER 1. STUDY ON THE REPETITIVE WEIGHT DROP MODEL OF TRAUMATIC BRAIN INJURY 18
SUMMARY 18
INTRODUCTION 19
MATERIALS AND METHODS 21
Experiment 1. Effect of SP-8356 on Repetitive WD TBI (2x) Model 21
Experiment 2. Effect of LMT-1154 on Repetitive WD TBI (2x) Model 23
Experiment 3. Effect of Repetitive WD TBI (7x) on Sensorimotor and Affective Behavior 27
Experiment 4. Effect of SP-8356 on Symptoms after Repetitive WD TBI (7x) 29
RESULTS 33
Experiment 1. Effect of SP-8356 on Repetitive WD TBI (2x) Model 33
Experiment 2. Effect of LMT-1154 on Repetitive WD TBI (2x) Model 34
Experiment 3. Effect of Repetitive WD TBI (7x) on Sensorimotor and Affective Behavior 35
Experiment 4. Effect of SP-8356 on Symptoms after Repetitive WD TBI (7x) 37
DISCUSSION 39
REFERENCES 60
CHAPTER 2. STUDY ON THE CONTROLLED CORTICAL IMPACT MODEL OF TRAUMATIC BRAIN INJURY 64
SUMMARY 64
INTRODUCTION 65
MATERIALS AND METHODS 67
Experiment 1. Effect of SP-8356 on the Controlled Cortical Impact Model of TBI 67
Experiment 2. Effect of LMT-1154 on the Controlled Cortical Impact Model of TBI 70
RESULTS 74
Experiment 1. Effect of SP-8356 on the Controlled Cortical Impact Model of TBI 74
Experiment 2. Effect of LMT-1154 on the Controlled Cortical Impact Model of TBI 75
DISCUSSION 78
REFERENCES 91
CHAPTER 3. EFFECT OF LIPOPOLYSACCHARIDE INJECTION ON TRAUMATIC BRAIN INJURY 96
SUMMARY 96
INTRODUCTION 97
MATERIALS AND METHODS 99
RESULTS 105
DISCUSSION 108
REFERENCES 125
Table 1.1.1. The sequence of primers for RT-qPCR 42
Table 2.1.1. The sequence of primers for RT-qPCR 80
Table 2.2.1. Neurological severity score for mice with head injuries 81
Table 3.1. The sequence of primers for RT-qPCR 112
Figure 1.1.1. Effect of SP-8356 on sensorimotor function following two repetitive weight drop injuries. In the rotarod test, the time point of falling from the rod or rotating the rod... 43
Figure 1.1.2. Effect of SP-8356 on inflammatory cytokines following two repetitive weight drop injuries. In RT-qPCR, detected levels of mRNA were normalized with GAPDH. Note... 44
Figure 1.1.3. Effects of SP-8356 on the CD147/MMP pathway in two repetitive weight drop injuries. In RT-qPCR, detected levels of mRNA were normalized with GAPDH. Note the... 45
Figure 1.2.1. Effect of LMT-1154 on body weight changes following two repetitive weight drop injuries. ▼, the time point of drug injection. Values are means for the six mice in the... 46
Figure 1.2.2. Effect of LMT-1154 on the loss of righting reflex following two repetitive weight drop injuries. The duration of LORR was defined as the time it takes for the four feet... 47
Figure 1.2.3. Effect of LMT-1154 on sensorimotor function following two repetitive weight drop injuries. In the rotarod test, the time point of falling from the rod or rotating the rod... 48
Figure 1.2.4. Effect of LMT-1154 on astrocyte and microglial activation following two repetitive weight drop injuries. From the ROI of the immunofluorescence staining, the... 49
Figure 1.3.1. Effect of seven repetitive weight drop injuries on changes in body weight. The mice's body weights in the sham and rmTBI groups were measured before the first injury, 1,... 50
Figure 1.3.2. Effect of seven repetitive weight drop injuries on sensorimotor function. In the rotarod test, the time point of falling from the rod or rotating the rod three times in a row... 51
Figure 1.3.3. Effect of seven repetitive weight drop injuries on locomotor and anxiety. In the open-field test, the mice were put into a corner of the open-field chamber and given 10... 52
Figure 1.3.4. Effect of seven repetitive weight drop injuries on despair-like behavior. (a) Immobilization time in the tail suspension test. (b) Increased immobilization time and (c)... 53
Figure 1.3.5. Effect of seven repetitive weight drop injuries on anxious behavior. In the elevated-0-maze test, (a) the entries, (b) the time spent in the open areas, and (c) the latency... 54
Figure 1.4.1. Effect of SP-8356 on body weight changes following seven repetitive weight drop injuries. Values are means for 11 mice in the sham-vehicle group, 14 in the TBI-vehicle... 55
Figure 1.4.2. Effect of SP-8356 on sensorimotor function following seven repetitive weight drop injuries. In the rotarod test, the time point of falling from the rod or rotating the rod... 56
Figure 1.4.3. Effect of SP-8356 on locomotor and anxiety following seven repetitive weight drop injuries. In the open field test, the mice were put into a corner of the open field chamber... 57
Figure 1.4.4. Effect of SP-8356 on despair-like behavior following seven repetitive weight drop injuries. (a) The immobilization time in the tail suspension test. (b) Immobilization... 58
Figure 1.4.5. Effect of SP-8356 on anxious behavior following seven repetitive weight drop injuries. In the elevated-0-maze test, (a) the entries, (b) the time spent in the open areas, and... 59
Figure 2.1.1. Effect of SP-8356 on body weight changes following CCI injury. ▼, the time point of drug injection. Values are means ± SD for 12 mice in the sham injury group and ten... 82
Figure 2.1.2. Effect of SP-8356 on impaired limb usage following CCI injury. The mice were placed in a cylinder for five minutes, and the number of instances the mice used each paw... 83
Figure 2.1.3. Effect of SP-8356 on inflammatory cytokines following CCI injury. In RT-qPCR, detected levels of mRNA were normalized with GAPDH. Note that SP-8356 does... 84
Figure 2.1.4. Effects of SP-8356 on the CD147/MMP pathway in CCI injury. (a) Quantitative analysis of the relative mRNA levels of CD147, MMP-2, and MMP-9 genes.... 85
Figure 2.2.1. Effect of LMT-1154 on body weight changes following CCI injury. Note that the body weight of both groups decreases until the second day after CCI and then rebounds... 86
Figure 2.2.2. Effect of LMT-1154 on sensorimotor function following CCI injury. In the rotarod test, the time point of falling from the rod or rotating the rod three times in a row... 87
Figure 2.2.3. Effect of LMT-1154 on neurological function following CCI. A neurological severity score (NSS) was used to assess neurological function. Ten items, including general... 88
Figure 2.2.4. Effect of LMT-1154 on impaired limb usage following CCI. Before inducing CCI injury and 1, 2, 3, 7, 10, and 14 days after the CCI injury, the number of instances the... 89
Figure 2.2.5. Effect of LMT-1154 on astrocyte and microglial activation following CCI injury. From the ROI of the immunofluorescence staining, the activated astrocytes were... 90
Figure 3.1. Effect of lipopolysaccharide pre-injection on body weight changes after weight drop injury. Body weight was measured at baseline before the LPS pre-injection, 24 h after... 113
Figure 3.2. Immunohistochemical staining of Iba-1 at 5 and 48 hours after TBI. The area of Iba-1 positive cells in the striatum and cortex among the ROI images was quantified. Each... 115
Figure 3.3. Immunohistochemical staining of CD68 at 5 and 48 hours after TBI. The area of CD68-positive cells in the striatum and cortex among the ROI images was quantified. Each... 117
Figure 3.4. Quantitative analysis of the relative mRNA levels between the saline-TBI and LPS-TBI groups. In RT-qPCR, detected levels of mRNA were normalized with GAPDH.... 118
Figure 3.5. Effect of lipopolysaccharide pre-injection on the CD147/MMP pathway. (a) Quantitative analysis of the relative mRNA levels of CD147, MMP-2, and MMP-9 genes.... 119
Figure 3.6. Effect of lipopolysaccharide pre-injection on sensorimotor function after weight drop injury. In the rotarod test, the time point of falling from the rod or rotating the rod three... 120
Figure 3.7. Effect of lipopolysaccharide pre-injection on locomotor and anxiety after weight drop injury. In the open field test, the mice were put into a corner of the open field chamber... 121
Figure 3.8. Effect of lipopolysaccharide pre-injection on anxious behavior after weight drop injury. In the elevated-0-maze test, (a) the entries, (b) the time spent in the open areas, and... 122
Figure 3.9. Effect of lipopolysaccharide pre-injection on despair-like behavior after weight drop injury. A tail suspension test was performed for 6 minutes, and the immobilization time... 123
Figure 3.10. Effect of lipopolysaccharide pre-injection on spatial working memory after weight drop injury. Mice were allowed to freely explore three pathways in a Y-shaped... 124