Protection of Chebulic Acid against Alveolar Epithelial Damage induced by Urban Particulate Matter
- 주제(키워드) Urban Particulate Matter , 미세먼지 , lung , chebulic acid
- 발행기관 고려대학교 대학원
- 지도교수 이광원
- 발행년도 2016
- 학위수여년월 2016. 8
- 학위구분 석사
- 학과 대학원 생명공학과
- 원문페이지 53 p
- 실제URI http://www.dcollection.net/handler/korea/000000068322
- 본문언어 영어
- 제출원본 000045881506
초록/요약
Urban Particulate Matter (SRM1648a) is atmospheric particulate matter collected in an urban area. Exposure to particulate matter is a significant risk for increased cardiopulmonary vascular morbidity and mortality. We studied the protective effect of chebulic acid (CA) against PM disrupting physical integrity of the pulmonary alveolar epithelial (PAE) monolayer. Barrier integrity of PAE depends upon the intercellular junction properties particularly tight junction, which consist of three types of transmembrane proteins. Human PAE cell line (NCI-H441) pretreated with CA for 24 h, and treated with PM. We verified noncytotoxicity of PM and CA using MTT assay. PM induced timedependent reactive oxygen species generation in alveolar epithelial cell as measurements by 2′,7′-Dichlorofluorescin diacetate (DCFH-DA). Furthermore, we showed that effect of PM on expression of junction protein RNA and protein levels. To analyze changes in integrity of PAE monolayer, we use transepithelial electrical resistance (TEER) measurement. And also, we measured paracellular permeability using the fluorescent tracer. These data showed that pretreatment of CA on alveolar epithelial cell reduced PM-induced ROS production, and protect disorganization of tight junction of PAE cells.
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CONTENTS
LIST OF FIGURE III
LIST OF TABLE 4
LITERATURE REVIEW 5
1.Terminalia chebula Retz. (T. chbula) 5
ABSTRACT 6
I. INTRODUCTION 7
II. MATERIALS AND METHODS 9
1. Materials 9
2. Preparation of Urban Particulate Matter (UPM) 10
3. Cell Culture 11
4. Cell Viability Assay 12
5. Measurement of Reactive Oxygen Species 13
6. Confocal Microscopy and Image Evaluation 14
7. ZO-1 Protein Staining 15
8. Western Blot Analysis 16
9. RNA Isolation and Quantitative real time PCR 17
10. Transepithelial Electrical Resistance (TEER) Measurement 19
11. Paracellular Permeability Assay 20
12. Statistical analysis 21
III. RESULTS 22
1. Cell viability of CA and UPM 22
2. ROS scavenging activities of CA 24
3. Inflammatory cytokines mRNA expression levels of PAE cells 26
4. Inflammatory cytokines mRNA expression levels of THP-1 cells 28
5. Tight junction mRNA and protein expression levels 30
6. ZO-1 immunofluorescence staining 32
7. Transmembrane epithelial electric resistance (TEER) 34
8. Paracellular permeability assay using fluorescently labelled dextran 36
IV. DISCUSSION 38
ABSTRACT IN KOREAN 43
REFERENCES 45
