鎳奈米粒子對子宮頸癌細胞與猴腎細胞之細胞毒性、氧化傷害及基因毒性

Cytotoxicity, Oxidative Stress and Genotoxicity of Nickel-nanoparticles in HeLa Cell and Vero Cell

林羿廷(Yi-Ting Lin)

指導教授 ： 黃友利

高雄醫學大學/健康科學院/生物醫學檢驗學研究所/碩士(2007年)

摘要

鎳奈米粒子具有高效催化劑、高效助燃劑等功能，被廣泛運用在工業材料上。雖然已有文獻證實鎳奈米粒子在血管內皮細胞和神經細胞無毒性反應，但鎳奈米粒子對其他細胞型態的毒性作用仍值得討論。本研究目的在於子宮頸癌細胞與猴腎細胞中探討鎳奈米粒子的細胞毒性、脂質過氧化作用及基因毒性。子宮頸癌細胞與猴腎細胞和各種鎳奈米粒子濃度 (0, 1, 5, 50, 100, 500 ?慊/mL培養液) ，共同培養48小時。本研究利用MTT分析細胞存活度 (cell viability)，評估細胞毒性；使用高效液相層析法測定丙二醛 (MDA)，探討脂質過氧化作用；利用彗星分析法及流式細胞法測定DNA傷害及細胞週期，進而評估基因毒性；以倒立顯微鏡觀察細胞的形態及電子顯微鏡證實鎳奈米粒子是否進入細胞內；藉由電熱式原子吸收光譜儀，分析鎳離子的存在與否。另外，加入抗氧化劑麩胱甘肽 (glutathione) 評估鎳奈米粒子對細胞存活度和MDA生成的影響。結果顯示，經鎳奈米粒子處理的子宮頸癌細胞與猴腎細胞，其細胞存活度降低，MDA增加、DNA斷裂傷害。另外，經倒立顯微鏡及穿透式電子顯微鏡觀察，證實細胞型態被破壞且鎳奈米粒子被噬入細胞內；而經由電熱式原子吸收光譜儀的測定，證明鎳離子的存在；添加麩胱甘肽可降低MDA產生及細胞死亡。上述結果證明鎳奈米粒子對子宮頸癌細胞和猴腎細胞確有細胞毒性、氧化性傷害及基因毒性。

關鍵字

鎳奈米粒子 ； 細胞毒性 ； 脂質過氧化 ； 基因毒性

並列摘要

Nickel-nanoparticles (Ni-NPs) are widely employed in the industry as the catalyzer, combustion supporter, and electrode material. Although no significant apparent cytotoxicity of Ni-NPs were found in the endothelial cell and the Neuro-2A cell, the possible effects of Ni-NPs in the other cell types remain to be elucidated. The aim of this study was to evaluate the cytotoxicity, lipid peroxidation and genotoxicity of Ni-NPs in HeLa cell and Vero cell. HeLa cell and Vero cell were incubated in growth medium containing 0, 1, 5, 50, 100, 500 Ni-NPs ?慊/ml culture medium for 48 h. Cell viability was measured by MTT assay for the investigation of cytotoxicity. The malondialdehyde (MDA) was determined by HPLC with thiobarbituric acid-reaction substance assay for the evaluation of lipid peroxidation. The DNA damage was performed by comet assay and flow cytometry for the evaluation of genotoxicity and cell cycle, respectively. The addition of glutathione in both HeLa cell and Vero cell were used to evaluate the effects of cell viability and MDA. Our results showed that the decreased cell viability, increased MDA and increased DNA damage were observed in both HeLa cell and Vero cell after the administration of Ni-NPs. Furthermore, the reverse microscopy and the transmission electron microscopy revealed the broken cell morphology in high concentration of Ni-NPs and differences in subcellular localization of the particles, respectively. In addition, the DNA fragments appeared in both HeLa cell and Vero cell. The addition of glutathione in both HeLa cell and Vero cell decreased the cell death and the production of MDA. The results of decreased cell viability, increased MDA and increased DNA damage and the effects of glutathione after the administration of Ni-NPs demonstrated the cytotoxicity, oxidative stress and genotoxicity of Ni-NPs in HeLa cell and Vero cell.

並列關鍵字

Nickel nanoparticles ； Cytotoxicity ； Lipid peroxidation ； Genotoxicity

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