Publications
Detailed Information
Modulatory mechanisms on the inflammation and muscarinic receptor function in salivary gland epithelial cells : 타액선 상피세포 내 염증 및 무스카리닉 수용체 조절기전
DC Field | Value | Language |
---|---|---|
dc.contributor.advisor | 박경표 | - |
dc.contributor.author | 신용환 | - |
dc.date.accessioned | 2017-07-14T05:50:29Z | - |
dc.date.available | 2017-07-14T05:50:29Z | - |
dc.date.issued | 2014-08 | - |
dc.identifier.other | 000000021386 | - |
dc.identifier.uri | https://hdl.handle.net/10371/125215 | - |
dc.description | 학위논문 (박사)-- 서울대학교 대학원 : 치의과학과, 2014. 8. 박경표. | - |
dc.description.abstract | Salivary gland epithelial cells (SGEC) release several cytokines that play important roles in the inflammatory process. Muscarinic receptors, particularly the type 3 subtype (M3R), play an important role in exocrine secretion. However, the regulatory mechanism of the inflammation and the functional expression of M3R in SGEC remains to be elucidated.
In chapterⅠ, I examined whether capsaicin can modulate the cytokine release in SGEC. These findings demonstrated that the increases in TNFα and IL-6 mRNA transcripts were highest at 3h and 1h after incubation with poly(I:C) and LPS, respectively. Pretreatment of the cells with 10 μΜ capsaicin, however, significantly inhibited mRNA transcripts and its protein levels. The simultaneous application of 10 μΜ capsazepine with capsaicin did not block the inhibitory effect of capsaicin. Furthermore, the inhibitory effect of capsaicin was also shown in primary cultured cells from TRPV1−/− mice. I found that both poly(I:C) and LPS induce IκB-α degradation and phosphorylation, which results in NF-κB activation and capsaicin inhibits this NF-κB pathway. These results demonstrate that SGEC release pro-inflammatory cytokines by TLR stimulation, and capsaicin inhibits this process by inhibiting the NF-κB pathway. In chapter Ⅱ, I examined A253 cells derived from human salivary gland tumor tissue, in which muscarinic receptor function is suppressed. In this study, I examined whether M3R function is suppressed by epigenetic modulation of the receptor. I found that A253 cells expressed all subtypes of muscarinic receptors, except subtype 3, at the mRNA and protein level. However, treatment of cells with 5-aza-2'-deoxycytidine (5-Aza-CdR) restored the functional expression of the M3R. Treatment of cells with 5-Aza-CdR completely restored the carbachol-induced calcium response, which was not observed in untreated A253 cells. Global methylation levels in A253 cells were also reduced by 5-Aza-CdR-treatment. I also examined whether 5-Aza-CdR treatment induced demethylation of the M3R CpG island, and found that one of the methylated CGs was demethylated by bisulfite sequencing. Thus, I conclude that suppression of M3R function in A253 cells results from hypermethylation of the CpG island | - |
dc.description.abstract | moreover, M3R function can be restored by DNA demethylation.
This study suggests that capsaicin could potentially alleviate the inflammation and 5-Aza-CdR could potentially be used to restore function to the M3R, which is suppressed in salivary gland epithelial cells. | - |
dc.description.tableofcontents | Contents
Abstract ⅰ Contents ⅲ List of Figures ⅴ Abbreviations vi General introduction 1 ChapterⅠ. Capsaicin regulates the NF-κB pathway in salivary gland inflammation 1. Introduction 6 2. Materials and Methods 8 3. Results 13 4. Discussion 25 ChapterⅡ. Epigenetic modulation of the muscarinic type 3 receptor in salivary epithelial cells 1. Introduction 29 2. Materials and Methods 31 3. Results 36 4. Discussion 53 References 56 Abstract in Korean 70 List of Figures ChapterⅠ Figure 1. TNFα and IL-6 mRNA levels in HSG cells stimulated with poly(I:C) or LPS 14 Figure 2. Expression of TRPV1 and application of capsaicin 17 Figure 3. Effect of capsaicin on TNFα and IL-6 mRNA expression and protein level in poly(I:C) and LPS-stimulated HSG cells 20 Figure 4. Inhibitory effect of capsaicin in TRPV1−/− mice and IκB-α/NF-κB signaling pathway 23 ChapterⅡ Figure 1. Induction of the M3R in A253 cells by treatment with 5-Aza-CdR 37 Figure 2. Induction of M3R mRNA expression and protein production by 5-Aza-CdR 40 Figure 3. Immunocytochemical staining of the M3R before and after 5-Aza-CdR treatment 43 Figure 4. Rescue of M3R function by 5-Aza-CdR treatment 45 Figure 5. Methylation levels in total DNA and the M3R CpG island upon treatment with 5-Aza-CdR 48 Figure 6. Bisulfite sequencing of the M3R CpG island before and after 5-Aza-CdR treatment 51 | - |
dc.format | application/pdf | - |
dc.format.extent | 1496877 bytes | - |
dc.format.medium | application/pdf | - |
dc.language.iso | en | - |
dc.publisher | 서울대학교 대학원 | - |
dc.subject | salivary gland | - |
dc.subject | capsaicin | - |
dc.subject | TNFα | - |
dc.subject | IL-6 | - |
dc.subject | capsazepine | - |
dc.subject | NF-κB | - |
dc.subject | epigenetic | - |
dc.subject | muscarinic receptor | - |
dc.subject | A253 | - |
dc.subject | hypermethylation | - |
dc.subject | 5-Aza-CdR | - |
dc.subject.ddc | 617 | - |
dc.title | Modulatory mechanisms on the inflammation and muscarinic receptor function in salivary gland epithelial cells | - |
dc.title.alternative | 타액선 상피세포 내 염증 및 무스카리닉 수용체 조절기전 | - |
dc.type | Thesis | - |
dc.description.degree | Doctor | - |
dc.citation.pages | vi, 71 | - |
dc.contributor.affiliation | 치과대학 치의과학과 | - |
dc.date.awarded | 2014-08 | - |
- Appears in Collections:
- Files in This Item:
Item View & Download Count
Items in S-Space are protected by copyright, with all rights reserved, unless otherwise indicated.