The Regulation Effects of Hesperetin on Hypersensitivity in Human Mast Cells and Microglia Cells
- Issued Date
- 2018
- Abstract
- IABSTRACT
The Regulation Effects of Hesperetin on Hypersensitivity
in Human Mast Cells and Microglia Cells
Jo, Sun hyo
Advisor : Prof. Lee, Jun Sik, Ph.D.
Department of Life Science,
Graduate School of Chosun University
Immune response is an essential defense mechanism to protect our bodies. However, inadequate immune response causes hypersensitivity leading to various diseases. Hypersensitivity is divided into four types. The present study focuses on type I and type IV hypersensitivity. Type I hypersensitivity is an allergic response and induced by histamine and cytokines released from mast cells. Atopic dermatitis is typical chronic inflammatory disease skin caused by a persistent allergic reaction. Type IV hypersensitivity is a inflammatory reactions caused by phagocytes such as macrophage and microglia. Neuroinflammation is a specific or nonspecific immunological reaction in the central nervous system that is distinct from peripheral inflammatory responses. Neuroinflammation is also induced by activation of microglial cells that acts as resident phagocytes in the brain. Activated microglial cells release a variety of pro-inflammatory mediator that lead to neuronal cell death and neurodegenerative diseases, such as Alzheimer’s and Parkin-son’s disease.
Hesperetin is one of the flavanones and is present in the peel of citrus. Hesperetin is reported to have anti-inflammatory, anti-cancer, and antioxidant effects. However, the anti-allergic and anti-neuroinflammatory effects of hesperetin on human mast cells and microglia cells are still unknown. Therefore, this study focused on the regulatory effects of hespretin on hypersensitivity through modulation of immune cells.
In the study of Part I, I examined whether hesperetin regulates the allergic inflammatory re-sponse in activated human mast cells. I found that hesperetin significantly inhibited gene expres-sion of pro-allergic inflammatory cytokine such as interleukin (IL)-6, TNF-α and IL-1β and secre-tion of IL-6 in PMA/A23187-stimulated human mast cells. In addition, I found that hesperetin inhibited the phosphorylation of the mitogen-activated protein kinase (MAPK) pathway including extracellular signal-regulated kinase (ERK) 1/2 and p38. Moreover, hesperetin suppressed atopic dermatitis and infiltration of mast cells in epidermis of DNCB-induced atopic dermatitis mouse model.
In Part II, I investigated the anti-neuroinflammatory effects of hesperetin on lipopolysaccharide (LPS)-stimulated BV-2 microglial cells. I found that hesperetin strongly inhibited nitric oxide (NO) production, as well as expression of inducible nitric oxide synthase (iNOS) in LPS-stimulated BV-2 microglial cells. Hesperetin also significantly reduced secretion of inflammatory cytokines, including IL-1β and IL-6. Furthermore, I found that hesperetin down-regulated the phosphorylation of ERK1/2 and p38 in MAPK pathway, resulting in anti-inflammatory effects. Moreover, Hesperetin suppressed astrocytes and microglial cells activation in LPS-challenged mice brain.
Therefore, my findings indicate that hesperetin inhibits allergic inflammation and neuroinflam-mation and has potential prophylactic application in treating allergic disease and neurodegenerative diseases by modulation of hypersensitivity mediated mast cells and microglia cells.
- Alternative Title
- Hesperetin에 의한 인간비만세포 및 신경아교세포의과민성 면역반응 조절 연구
- Alternative Author(s)
- Jo, Sun Hyo
- Affiliation
- 조선대학교 일반대학원 생명과학과
- Department
- 일반대학원 생명과학
- Advisor
- 이준식
- Awarded Date
- 2019-02
- Table Of Contents
- CONTENTS
LIST OF TABLES................................................................i
LIST OF FIGURES.............................................................ii
ABBREVIATIONS.............................................................iii
ABSTRACT..........................................................................v
국문초록.............................................................................vii
I. INTRODUCTION............................................................1
1. Hypersensitivity in immune response.......................................1
A. Type I hypersensitivity...............................................................2
(1) Mast cells...............................................................................................2
(2) Atopic dermatitis...................................................................................5
B. Type II of hypersensitivity............................................................6
C. Type III of hypersensitivity............................................................8
D. Type IV of hypersensitivity...........................................................10
(1) Neuroinflammatory response............................................................10
2. Hesperetin...................................................................................15
3. Inflammatory signaling pathway…….....................................17
A. Mitogen-activated protein kinase (MAPK) pathway................17
B. Nuclear factor kappa-B (NF-κB) pathway.................................18
4. Purpose of study............................……..................................21
II. MATERIALS AND METHODS.................................22
1. Mice........................................................................................22
2. Chemicals and reagents........................................................22
3. Cell culture and chemical treatment..................................22
4. Annexin V & PI staining......................................................23
5. Cytotoxicity assay..................................................................23
6. NO assay.................................................................................23
7. Reverse transcription (RT)-PCR.........................................24
8. Western blot analysis............................................................26
9. Enzyme-linked immunosorbent assay (ELISA).................27
10. Animal model........................................................................27
11. Histological analysis..............................................................28
12. Immunohistochemistry.........................................................28
13. Statistical analysis.................................................................29
III. RESULTS.....................................................................31
Part I. Anti-allergic effect of hesperetin on human mast cell activation via MAPK pathway regulation.
1. Hesperetin had no effect on cytotoxicity in human mast cells................................................................................................31
2. Hesperetin suppresses the gene expression levels of pro-inflammatory cytokines in human mast cell activation............33
3. Hesperetin significantly inhibits allergic inflammatory cytokine production in human mast cell activation...............................35
4. Hesperetin inhibits phosphorylation of ERK and p38 MAPK pathway in activated human mast cells............................................37
5. Hesperetin attenuates epidermal hypertrophy and mast cell infiltration in DNCB-induced atopic dermatitis........................39
Part II. Hesperetin regulates LPS-induced neuroinflammation on microglia by suppressing pro-inflammatory cytokines and MAPK phosphorylation.
1. Hesperetin inhibits NO production in LPS-stimulated BV-2 microglial cells..............................................................................41
2. Hesperetin inhibits the expression of pro-inflammatory cytokines and iNOS in LPS-stimulated BV-2 microglial cells..43
3. Hesperetin reduces the production of pro-inflammatory cytokine such as TNF-α, IL-1β, and IL-6.................................46
4. Hesperetin inhibits phosphorylation of p38 MAPK and ERK in LPS-stimulated BV-2 microglial cells.........................................49
5. Hesperetin attenuates astrocyte and microglia activation in the LPS-administrated mouse brain.................................................50
IV. DISCUSSION...............................................................53
V. REFERENCES............................................................57
감사의 글...........................................................................64
- Degree
- Master
- Publisher
- 조선대학교 일반대학원 생명과학과
- Citation
- 조순효. (2018). The Regulation Effects of Hesperetin on Hypersensitivity in Human Mast Cells and Microglia Cells.
- Type
- Dissertation
- URI
- https://oak.chosun.ac.kr/handle/2020.oak/13723
http://chosun.dcollection.net/common/orgView/200000267084
- Appears in Collections:
- General Graduate School > 3. Theses(Master)
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