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Effects of 4-Hexylresorcinol on the Reactive Oxygen Species induced by Oxidative Stress in Dental Pulp Cells of Rats : 흰 쥐의 치수세포에서 4-Hexylresorcinol 투여가 산화스트레스로 인한 활성산소에 미치는 영향

DC Field Value Language
dc.contributor.advisor김태우-
dc.contributor.author장준호-
dc.date.accessioned2022-06-08T06:42:56Z-
dc.date.available2022-06-08T06:42:56Z-
dc.date.issued2022-
dc.identifier.other000000169671-
dc.identifier.urihttps://hdl.handle.net/10371/181240-
dc.identifier.urihttps://dcollection.snu.ac.kr/common/orgView/000000169671ko_KR
dc.description학위논문(박사) -- 서울대학교대학원 : 치과대학 치의과학과, 2022.2. 김태우.-
dc.description.abstractObjective: It is crucial for dental pulp to manage the stress avoiding wound healing delay and consequent inflammation. The application of anti-oxidant may be beneficial preventing the such complications. In this study, we aimed to evaluate the therapeutic effects of 4- Hexylresorcinol (4HR), one of the well-known antioxidants, on dental pulp cells.

Methods: In cellular experiments, dental pulp cells from rat mandibular incisor were cultured and treated with 4HR (1–100 µM). Changes in total antioxidant capacity (TAC) and glutathione peroxidase (GPx) activity in response to 4HR treatment were evaluated following hydrogen peroxide treatments. Primary antibodies against tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were used in evaluation assays. The positive control was H2O2 treatment without 4HR pretreatment, whereas the negative control was no treatment group.
In animal experiment, twenty rats were subjected to the experiment. Two rats were used to measure the baseline activity of TAC and GPx after pulp tissue extraction. Eighteen rats were devided into two groups. The experimental group (n = 9) was treated with 2% 4HR ointment and the control group (n = 9) treated with the ointment base (lanolin) only. Three animals (n = 3 / 9) were used for protein extraction and Western blot analysis 3 days after the application. And six animals (n = 6 / 9) were used for histological and immunohistochemical analyses (n = 3) and for TAC and GPx analyses (n = 3) 5 days after the application.

Results: TAC and GPx activities were significantly increased (p < 0.05), while the expression levels of TNF-α and IL-1β were decreased following the exposure to 4HR pretreatment in an in vitro model (p < 0.05). Additionally, the application of 4HR ointment in an exposed dental pulp model significantly reduced the expression of TNF-α and IL-1β (p < 0.001).

Conclusions: 4HR exerted protective effects against oxidative stress in dental pulp tissues through downregulating TNF-α and IL-1β.
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dc.description.abstract1. 목 적
외상이나 교정력에 의해 치아는 스트레스를 받게 되며, 이에 대한 생물학적 반응으로 발생하는 산화 스트레스(oxidative stress)는 tumor necrosis factor-α (TNF-α)와 interleukin-1β (IL-1β) 같은 염증 촉진(pro-inflammatory) 요인들을 증가시켜 구강내 염증 반응을 일으킨다. 4-Hexylresorcinol (4HR)은 resveratrol과 유사한 항산화제로 밝혀졌으며, 항산화제를 투여하여 과도한 산화 스트레스를 경감시키는 것은 치수조직의 치유에 도움을 줄 수 있다. 본 연구의 목적은 치수에서 산화스트레스에 대한 항산화제로서4HR의 치료 효과를 평가하는 것이다.

2. 방 법
생체 외(in vitro) 실험을 위하여 흰쥐의 하악 전치에서 치수 조직을 채취하여 배양한 dental pulp cell에 다양한 농도의 4HR (1,10,100 µM)을 전처리하고 H2O2를 처치한 후 total antioxidant capacity (TAC)와 glutathione peroxidase (GPx) 활성도를 측정하였다. TNF-α와 IL-1β의 항체에 대한 western blot을 시행하여 단백질 발현정도를 평가하였다.
생체 내(in vivo) 실험으로는 18마리(실험군, 대조군 각 9마리)의 흰 쥐에 하악 우측 절치를 절단하고 각각 2% 4HR ointment, ointment base (lanolin)를 도포하였다. 희생 후 TNF-α와 IL-1β에 대한 western blot을 시행하였으며 면역조직화학분석(immunohistochemical analysis) 및 TAC 와 GPx 활성도를 측정하였다. 하악 절치를 절단하지 않은 2마리는 TAC 와 GPx의 baseline activity를 측정하는데 이용하였다.

3. 결 과
1) 생체 외(in vitro) 실험: 4HR 투여 시 치수배양세포에서TAC 와 GPx 활성도가 증가하였으며 (p < 0.05), TNF-α 와 IL-1β 발현 수치는 감소하였다 (p < 0.05).
2) 생체 내(in vivo) 실험: 외상에 의한 스트레스를 받은 치수조직에 4HR ointment 적용시 산화 스트레스를 완화시키며, TNF-α 와 IL-1β 발현 수치 역시 감소하였다 (p < 0.001).

4. 결 론
이상의 결과를 바탕으로, 4HR 투여가 TNF-α와 IL-1β를 감소시켜 산화스트레스로부터 치수 조직을 보호하는 효과가 있다고 판단된다.
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dc.description.tableofcontentsI. INTRODUCTION 1
II. REVIEW OF LITERATURE
1. Reactive oxygen species and oxidative damage 3
2. Oxidative stress in orthodontic treatment 5
3. Pulpal reaction to orthodontic force 7
4. Biological properties and applications of 4HR 8
III. MATERIALS AND METHODS
Part 1. Cellular experiments
1.1. Cell culture and 4HR treatment 11
1.2. Assessing GPx Activity and TAC 11
1.3. Evaluating TNF-α and IL-1β Expression in Dental Pulp Cells after 4HR Treatment 12
Part 2. Animal experiments
2.1. Animals and experimental design 14
2.2. Histological, Immunohistochemical, Western Blot, GPx activity and TAC Analysis 14
Part 3. Statistical analysis 16
IV. RESULTS
Part 1. Cellular experiments
1.1. Application of 4HR Increases GPx Activity and TAC in Dental Pulp Cells 17
1.2. 4HR Decreases the TNF-α and IL-1β Expression Induced by Hydrogen Peroxidase 18
Part 2. Animal experiments
2.1. 4HR Decreased Inflammatory Reaction of Pulp Tissue Induced by Physical Stress 20
V. DISCUSSION 22
VI. CONCLUSIONS 27
REFERENCES 28
TABLES AND FIGURES 37
ABSTRACT IN KOREAN 46
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dc.format.extent47-
dc.language.isoeng-
dc.publisher서울대학교 대학원-
dc.subject4-Hexylresorcinol, reactive oxygen species, dental pulp, oxidative stress-
dc.subject.ddc617.6-
dc.titleEffects of 4-Hexylresorcinol on the Reactive Oxygen Species induced by Oxidative Stress in Dental Pulp Cells of Rats-
dc.title.alternative흰 쥐의 치수세포에서 4-Hexylresorcinol 투여가 산화스트레스로 인한 활성산소에 미치는 영향-
dc.typeThesis-
dc.typeDissertation-
dc.contributor.AlternativeAuthorChang Jun-Ho-
dc.contributor.department치과대학 치의과학과-
dc.description.degree박사-
dc.date.awarded2022-02-
dc.identifier.uciI804:11032-000000169671-
dc.identifier.holdings000000000047▲000000000054▲000000169671▲-
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