- 學位論文
探討1-Nitropyrene誘導視網膜色素上皮細胞氧化壓力及細胞凋亡之相關機制
Mechanism of 1-nitropyrene induced oxidative stress and apoptosis in ARPE-19 cells
摘要
隨著科技日新月異，空氣汙染越來越嚴重，形成一個重要的環境議題。空氣汙染的成分非常多種，如：二氧化氮、臭氧、二氧化硫和多環芳香烴化合物等成分，其中多環芳香烴化合物又以硝基多環芳香烴中的 1-Nitropyrene (1-NP) 最具代表性。有文獻指出，硝基多環芳香烴具有高度的誘發突變性，不必經代謝作用，已具生物活性，能誘發原核細胞和真核細胞發生基因突變，造成細胞損傷。因此本實驗研究目的是探討 1-NP 是否會進入人體內，造成眼睛內的視網膜傷害，以及其相關的傷害機制討論。 結果發現，1-NP 作用24小時後，會誘發視網膜色素上皮細胞 (adult retinal pigment epithelial-19, ARPE-19) 產生氧化壓力，造成細胞內的超氧化物歧化酶 (superoxidase dismutase，SOD) 、過氧化氫酶 (catalase) 、穀胱甘肽 (glutathione) 等等的抗氧化物的活性減少，造成細胞受損。另一方面，1-NP 作用後所產生的氧化壓力，也會導致 ARPE-19 細胞產生脂質過氧化的現象，讓細胞內的丙二醛 (MDA) 表現量增加，造成細胞受到傷害。此外，透過 MTT、LDH 以及 AnnexinV/PI 這三種實驗可觀察到，1-NP 會對 ARPE-19 細胞產生細胞毒性。最後，透過 western blot 實驗發現 1-NP 會磷酸化 p38 MAPK 、 ERK 以及 p53 造成細胞凋亡。此外，還發現 1-NP 會磷酸化 NFκB pathway 的相關蛋白 NFκB 以及 IκB-α，進而導致下游 iNOS 以及 COX-2 活化，最終導致細胞產生發炎反應，另外還發現 1-NP 會透過磷酸化 Nrf2 以及降解 Keap1 造成細胞產生氧化反應。本論文確定 1-NP 會對 ARPE-19 細胞產生細胞毒性，造成氧化壓力、細胞凋亡以及發炎反應。
並列摘要
With the rapid development of technology, air pollution is becoming more and more serious, forming an important environmental issue. There are many kinds of air pollution components, such as nitrogen dioxide, ozone, sulfur dioxide and polycyclic aromatic hydrocarbon compounds. Among this air pollution components, 1-Nitropyrene (1-NP) are representative in nitropolycyclic aromatic hydrocarbons. It has been pointed out in the literature that nitropolycyclic aromatic hydrocarbons have a high degree of induced mutation. It are not biologically active, and can induce gene mutations in prokaryotic cells and eukaryotic cells, resulting in cell damage. Therefore, the purpose of this experimental study is to investigate whether 1-NP will enter the human body, causing retinal damage in the eye and its associated injury mechanism. It was found that after 24 hours of 1-NP treatment, oxidative stress was induced in the retinal pigment epithelial cells (ARPE-19), resulting in superoxide dismutase (SOD) in the cells. The activity of antioxidants such as catalase, glutathione, etc. is reduced, causing cell damage. On the other hand, the oxidative stress generated by the action of 1-NP also causes lipid peroxidation in ARPE-19 cells, which increases the amount of malondialdehyde (MDA) in the cells and causes damage to cells. In addition, 1-NP was observed to be cytotoxic to ARPE-19 cells by three experiments, MTT, LDH and AnnexinV/PI. Finally, wesrern blot analysis revealed that 1-NP phosphorylates p38 MAPK, ERK and p53 to cause apoptosis. In addition, 1-NP was also found to phosphorylate NFκB pathway-associated proteins NFκB and IκB-α, which in turn led to the activation of downstream iNOS and COX-2, which ultimately led to inflammatory responses in cells, and also found that 1-NP phosphorylates Nrf2 and Degradation of Keap1 causes oxidation of cells. This paper determined that 1-NP can cause cytotoxicity to ARPE-19 cells, resulting in oxidative stress, apoptosis and inflammatory response.
參考文獻
延伸閱讀
- Natalia, S. L. (2017). 氧化壓力誘導Musashi-1 斷裂造成視網膜色素上皮 細胞變性 [master's thesis, National Chiao Tung University]. Airiti Library. https://www.airitilibrary.com/Article/Detail?DocID=U0030-0705201811493452
- 呂偉台（2022）。探討 1-Undecanol 誘導人類角膜上皮細胞之氧化壓力、細胞焦亡及分子機轉〔碩士論文，中山醫學大學〕。華藝線上圖書館。https://doi.org/10.6834/csmu202200153
- 張集武（2007）。探討HMG CoA還原酶抑制劑對於視網膜色素上皮細胞分泌血管內皮生長因子的影響〔碩士論文，高雄醫學大學〕。華藝線上圖書館。https://www.airitilibrary.com/Article/Detail?DocID=U0011-2707200700270800
- 黃文郁（2007）。Investigation of Methotrexate-Induced Cell Cycle Arrest and Apoptosis〔碩士論文，國立臺灣大學〕。華藝線上圖書館。https://doi.org/10.6342/NTU.2007.02753
- 林俊祐（2009）。Inhibition of Endotoxin-Induced Nitric Oxide Synthase-2 Synthesis by Pheophytin a in Macrophages〔碩士論文，高雄醫學大學〕。華藝線上圖書館。https://doi.org/10.6832/KMU.2009.00014