The Effect of Trolox on Oxidative Damage in Human Skin Fibroblast Cells : The Effect of Trolox on Oxidative Damage in Human Skin Fibroblast Cells
- 발행기관 고려대학교
- 발행년도 2007
- 학위수여년월 2007. 8
- 학위명 박사
- 학과 대학원 생명공학과 의약생명공학전공
- 식별자(기타) DL:000018571595
- 서지제어번호 000045398052
초록/요약
본 연구는 human skin fibroblast cell들의 oxidative damage에 대한 Trolox의 antioxidative effect에 관한 내용이다. 특히AFM을 이용하여, oxidative stress동안 skin cell들의morphology와 surface feature의 변화 양상을 관찰할 뿐 아니라, 그것을 수치화하는 방법을 제시하였다. 또한 skin cell들의 oxidative damage에 대한 Trolox의 antioxidant effect의 action mechanism을 제시하였다. Human skin fibroblast cell 들의 cytoskeleton structure는 ROS/oxidative stress에 의해 비교적 초기 단계에 가장 쉽게 손상되는 대표적인 세포 성분 중 하나로 판단된다. Oxidative agent에 의한 cytoskeleton의 손상은human skin fibroblast cell 들의morphology 변화 및 cell death로 이어졌다. 그러나 Trolox는ROS에 의해 유발되는cytoskeleton의 손상 및roughness, dimension, stiffness, morphology변화를 차단하는 ROS/oxidative stress로 부터human skin fibroblast cell들을 보호하는 효과를 보였다. 이러한 Trolox의 antioxidative effect는 부분적으로 세포 내 ROS를 제거함으로써 진행됨이 확인되었다. 또한 증가된sub G1 population, PARP cleavage, MMP disruption, caspase-9 and -3 activities, Bax:Bcl-2 ratio increase와 같은 ROS에 노출된human skin fibroblast cell들에서 나타나는 대표적 apoptotic feature들을 완화시키는 효과를 나타냈다. 결론적으로, cytoskeleton의 oxidative damage가 노출의 비교적 초기 단계에 진행되며, 특히 그것의 변형은mitochondria-dependent pro-apoptotic signal을 포함하는 cell death cascade를 활성화 시킨다고 판단된다.
more초록/요약
The overall and nanoscale morphologies and mechanics of cytostructural elements of human skin fibroblast cells were examined during oxidative stress-induced skin damage. Human skin fibroblast cells exposed to oxidizing agents were imaged with atomic force microscopy (AFM), and their cytoskeletal organization, morphology, oxidative stress, and viability were marked at the following time points. AFM was also employed to evaluate quantitatively the apparent responses of human skin fibroblast cells to the oxidative stress signaling mechanism. The cytoskeleton structure of human skin fibroblast cells seemed to be one of the earliest and most vulnerable cellular components injured by reactive oxygen species (ROS)-induced oxidative stress. Oxidative agents induced time- and concentration-dependent disruption of native organization of cytoskeleton with a decrease in cell integrity. These dynamic changes in cell mechanical elements were accompanied by morphological changes, increased oxidative stress, and loss of cell viability. A water-soluble derivative of vitamin E, Trolox, showed a preventive effect on ROS-induced cell death and cytoskeleton loss accompanied with roughness, dimension, stiffness, and morphology changes. Furthermore, Trolox extended cytoskeleton structures, resulting in decreased roughness of skin cells. The Trolox effects were in part through repression of oxidative stress in skin cells by decreasing their intracellular ROS level. More importantly, Trolox attenuated several apoptotic features displayed in H2O2 -exposed human skin fibroblast cells, including increased sub G1 population, poly-(ADP-ribose) polymerase (PARP) cleavage, mitochondrial membrane potential (MMP) disruption, and caspase-9 and -3 activities in a dose-dependent manner. Furthermore, pretreatment of human skin fibroblast cells with Trolox not only increased the protein expression level of Bcl-2, but also decreased Bax after H2O2 addition. The results indicate that the oxidative damage to cytoskeleton might occur at relatively early stages of exposure. Furthermore, alterations of cellular constituents like microfilaments seem to activate the cell death cascade, most likely pro-apoptotic signals related to the mitochondria-dependent pathway.
more