Neuroprotective effect of β-lapachone against glutamate-induced injury in HT22 cells
- 주제(키워드) β-lapachone , glutamate , oxidative stress , neuroprotective effects , antioxidant effects
- 주제(DDC) 615.1
- 발행기관 아주대학교 일반대학원
- 지도교수 정이숙
- 발행년도 2025
- 학위수여년월 2025. 2
- 학위명 석사
- 학과 및 전공 일반대학원 약학과
- 실제URI http://www.dcollection.net/handler/ajou/000000034447
- 본문언어 한국어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Neuroprotective effect of β-lapachone against glutamate- induced injury in HT22 cells While glutamate, a key neurotransmitter in the central nervous system, is fundamental to neuronal viability and normal brain function, its excessive accumulation leads to oxidative stress, contributing to neuronal damage and neurodegenerative diseases. In this study, we investigated the effect of β-lapachone (β-Lap), a naturally occurring naphthoquinone, on glutamate-induced injury in HT22 cells and explored the underlying mechanism involved. Our results show that β-Lap significantly improved cell viability in a dose-dependent manner. Additionally, β- Lap exhibited a significant antioxidant activity, reducing levels of intracellular reactive oxygen species and restoring glutathione levels. The antioxidant capacity of β-Lap was further demonstrated through 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging assays. Western blot analysis revealed that β-Lap upregulated brain-derived neurotrophic factor (BDNF) and promoted the phosphorylation of tropomyosin receptor kinase B (TrkB), extracellular signal-regulated kinase (ERK), and cAMP response element-binding protein (CREB), which were downregulated by glutamate. Furthermore, β-Lap enhanced the cellular antioxidant molecules, nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1). In conclusion, β-Lap can protect HT22 cells against glutamate-induced injury by activating the BDNF/TrkB/ERK/CREB and ERK/Nrf2/HO-1 signaling pathways, suggesting its therapeutic potential for neurodegenerative diseases.
more목차
Ⅰ. INTRODUCTION 1
A. Neurodegenerative disease 1
B. Glutamate-induced oxidative stress 1
C. BDNF/TrkB signaling pathway 2
D. Nrf2/HO-1 signaling pathway 3
E. β-lapachone 3
F. Aims of study 4
Ⅱ. MATERIALS AND METHODS 5
A. Reagents 5
B. Cell culture 6
C. Cell viability assay 6
D. DPPH radical-scavenging assay 6
E. ABTS radical-scavenging assay 7
F. Measurement of reactive oxygen species 7
G. Estimation of intracellular glutathione 8
H. Nuclear and cytosolic protein extraction 8
I. Western blot analysis 9
J. Statistical analysis 9
Ⅲ. RESULTS 10
A. β-Lap exerts a neuroprotective effect against glutamate-induced cytotoxicity in HT22 cells 10
B. β-Lap exhibits antioxidant effects in glutamate-exposed HT22 cells 13
C. β-Lap exerts in vitro antioxidant activity 15
D. β-Lap activates the BDNF/TrkB/ERK/CREB signaling molecules 17
E. β-Lap upregulates Nrf2 nuclear translocation and the expression of antioxidant enzyme 20
F. ANA-12 inhibits the neuroprotective effects of β-Lap 23
G. U0126 inhibits the neuroprotective effects of β-Lap 30
IV. DISCUSSION 33
V. CONCLUSION 37
REFERENCES 38
