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Anti-cancer effects of Lapathoside A from Fagopyrum esculentum in human pancreatic cancer cells and neuroprotective properties of Ecklonia cava ferment extract against glutamate-induced oxidative stress in HT22 hippocampal cell

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Abstract
Lapathoside A, a phenylpropanoid ester compound, was isolated from buckwheat roots. Ethanol (70%) extract of buckwheat roots was separated into n-hexane, methylene chloride, ethyl acetate, n-butanol, and water fractions by solvent partitioning. Seven fractions were obtained from the ethyl acetate fraction by liquid chromatography. Of the seven fractions, fraction No. 6 contained lapathoside A. The structure of lapathoside A was determined by LC-MS, LC-MS/MS, and NMR analysis. Next, we investigated the anticancer activity of lapathoside A in human pancreatic cancer cell lines (PANC-1 and SNU-213). After treatment with lapathoside A at 25 M, viabilities μ of PANC-1 and SNU-213 cells decreased 40% and 27%, respectively. Treatment with lapathoside A also increased the number of apoptotic cells and regulated expression levels of apoptotic proteins. Results using flow cytometric analysis indicated that treatment with lapathoside A increased the number of apoptotic cells in pancreatic cancer cell lines. These results suggest that lapathoside A might have potential application of the treatment of pancreatic cancer.
Glutamate induces neuronal cell death and thought to be involved in advance of neurodegenerative diseases through Reactive Oxygen Species (ROS) generation. ROS is known to cause neurodegenerative diseases. Ecklonia cava is known to have biological activities such as antioxidant, anti-inflammatory, anticancer, antidiabetic effects. In the present study, protective effects of E. cava ethanol extract and E. cava ferment extract was investigated against glutamate-induced neuronal cell death through oxidative stress. Cell viability was measured using WST-1 assay and confirmed that EFE had no toxicity on HT22. And EFE protected HT22 cells from glutamate-induced cell death. Also, apoptotic cell death induced by glutamate was attenuated by EFE. Flow cytometric analysis for detection of intracellular ROS generation showed that EFE reduced glutamate-induced ROS production. These results suggest that EFE have neuroprotective effect by attenuates glutamate-induced apoptosis and ROS generation in HT22 cells.
Author(s)
Kang, Mi Sook
Issued Date
2020
Awarded Date
2020. 8
Type
Dissertation
URI
http://dcoll.jejunu.ac.kr/common/orgView/000000009726
Alternative Author(s)
강미숙
Affiliation
제주대학교 대학원
Department
대학원 생명공학과
Advisor
김재훈
Table Of Contents
PART Ⅰ
Lapathoside A isolated from Fagopyrum esculentum induces apoptosis
in human pancreatic cancer cells.
CONTENTS
CONTENTS ⅰ
LIST OF FIGURES ⅲ
LIST OF TABLES ⅳ
ABSTRACT 1
1. INTRODUCTION 2
2. MATERIALS AND METHODS 4
2.1. Extraction and isolation of lapathoside A from buckwheat roots 4
2.2. Identification of lapathoside A 4
2.3. Cell culture 5
2.4. Cell viability assay 5
2.5. Flow cytometric analysis for cell apoptosis detection 6
2.6. Flow cytometric analysis for Reactive Oxygen Species (ROS) detection 6
2.7. Western blot analysis 7
2.8. Statistical analysis 7
3. RESULTS 9
3.1. Isolation and identification of lapathoside A 9
3.2. Lapathoside A inhibits pancreatic cancer cell growth 14
3.3. Lapathoside A induces the apoptosis of PANC-1 and SNU-213 cells 16
3.4. Lapathoside A induces ROS generation in human pancreatic cancer cells 20
3.5. Effects of lapathoside A on the phosphorylation of FAK, Akt proteins and MAPKs 22
4. DISCUSSION 25
5. REFERENCES 27
PART Ⅱ
Ecklonia cava ferment extract attenuates glutamate-induced oxidative stress in HT22 hippocampal cell.
CONTENTS
CONTENTS ⅰ
LIST OF FIGURES ⅱ
ABSTRACT 1
1. INTRODUCTION 2
2. MATERIALS AND METHODS 4
2.1. Fermentation and extraction 4
2.2. Cell culture 4
2.3. Cell viability assay 4
2.4. Flow cytometric analysis for cell apoptosis detection 4
2.5. Flow cytometric analysis for ROS detection 5
2.6. Statistical analysis 5
3. RESULTS 6
3.1. EFE protects against glutamate-induced cytotoxicity 6
3.2. EFE protects against glutamate-induced apoptotic cell death 8
3.3. EFE prevents ROS production induced by glutamate 10
4. DISCUSSION 12
5. REFERENCES 13
ACKNOWLEDGEMENT 15
Degree
Master
Publisher
제주대학교
Citation
Kang, Mi Sook. (2020). Anti-cancer effects of Lapathoside A from Fagopyrum esculentum in human pancreatic cancer cells and neuroprotective properties of Ecklonia cava ferment extract against glutamate-induced oxidative stress in HT22 hippocampal cell
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Faculty of Biotechnology > Molecular Biotechnology
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