인간 치수세포에서 H2O2로 유도된 Reactive Oxygen Species 생성에 대한 Tat-Antioxidant 1 융합 단백질의 효과
Effect of Tat-Antioxidant 1 fusion protein on H2O2-induced reactive oxygen species production in Human dental pulp cells
- 주제(키워드) 도움말 Human dental pulp cells , Tat-Atox1 , Protein transduction domain , Reactive oxygen species
- 발행기관 강릉원주대학교
- 발행년도 2016
- 학위수여년월 2016. 2
- 학위명 석사
- 학과 및 전공 도움말 일반대학원 치의학과
- 세부전공 생화학 및 분자생물학
- 실제URI http://www.dcollection.net/handler/kangnung/000000008089
- 본문언어 한국어
초록/요약 도움말
Dental pulp is a mesenchymal tissue surrounded by dentin, a mineralized tissue, and it contains blood vessels and nervous tissue. The damage to dental pulp due to mechanical, chemical, and temperature stimuli as well as microbial stimuli causes various inflammatory responses. The inflammatory response of the dental pulp causes tissue breakdown, induces cell damage and activates the factors that induce inflammatory responses. H2O2 is a typical ROS and causes apoptosis of human dental pulp (HDP) cells. Antioxidant 1 (Atox1) controls copper homeostasis and promotes cellular antioxidation of the toxins produced by the superoxide anion and hydrogen peroxide. However, the role of Atox1 in dental pulp diseases is not well known. In this study, the anti-inflammation activity and anti-apoptotic effect of Tat-Atox1, which can penetrate cells, were tested by inducing inflammatory responses with H2O2 in HDP cells. Atox1 alone cannot penetrate a cell, and thus a Tat-Atox1 fusion protein was created using the protein transduction domain (PTD). The purified Tat-Atox1 fusion protein was transduced into HDP cells in a time- and concentration-dependent manner, and effective inhibition of H2O2 induced-apoptosis was confirmed through Western blot analysis. Tat-Atox1 increased the survival rate of cells containing H2O2 and reduced the ROS level, there by playing a protective role. This result indicates that Tat-Atox1 can be used for the inhibition of HDP apoptosis and as a agent for dental pulp diseases.
more목차 도움말
1.서론 1
2.재료및방법 4
2.1. 치수세포(humandentalpulp;HDP)준비 4
2.2. Tat-Atox1plasmid의발현 4
2.3. Tat-Atox1융합단백질의발현과정제 5
2.4. 치수세포에Tat-Atox1융합단백질의형질도입및H2O2처리 5
2.5. Westernblotanalysis 6
2.6. Confocalfluorescencemicroscope 7
2.7. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide(MTT)assay 7
2.8. 통계분석 8
3. 결과 9
3.1. Tat-Atox1융합단백질의발현과정제 9
3.2. 치수세포에서Tat-Atox1융합단백질의형질도입 13
3.3. H2O2로산화스트레스를유도한치수세포생존율에대한Tat-Atox1의효과 17
3.4. H2O2로산화스트레스를유도한치수세포에서Bcl-2와Bax발현의변화 19
3.5. H2O2로산화스트레스를유도한치수세포에서caspase의활성화변화 21
4.고찰 24
5.결론 29
6.참고문헌 30
7.영문초록 36
