FRTL-5 갑상선세포에서 Potassium Bromate 독성에 대한 항산화제의 보호효과

Other Titles
Protective Effects of Antioxidants on Potassium Bromate-Induced Cytotoxicity in FRTL-5 Thyroid Cells
Authors
도영민
Abstract
갑상선세포에서 일상생활에서 섭취 가능한 항산화물질들이 KBrO₃가 유발하는 독성에 대한 보호효과가 있는지 알아보고자 정상 흰쥐의 갑상선세포주인 FRTL-5를 이용하여 200 μM potassium bromate (KBrO₃)를 처리하여 세포독성을 유발하였다. KBrO₃에 의한 갑상선세포의 독성에 대해 항산화물질들의 효과를 관찰하기 위해 N-acetyl-cysteine (NAC), L-2-oxothiazoline-4 -carboxylate (OTC), L-N(G)-nitroarginine methylester (L-NAME), melatonin, D-methionine, minocycline, (-)-epigallocatechin-3-gallate (EGCG), retinol, ascorbic acid, α-tocopherol을 각각 2-24시간 전처치한 후 FRTL-5 세포의 생존율의 변화를 관찰하였다. NAC과 OTC을 각각 전처치한 후 KBrO₃에 의한 FRTL-5 세포의 생존율은 60%에서 약 95%까지 농도에 의존적으로 증가하였고, minocycline을 전처치 하였을 때 세포생존율은 70% 이상 증가하였다. 그러나 L-NAME를 전처치한 경우에 KBrO₃에 의한 FRTL-5 세포의 생존율에는 변화가 없었다. Ascorbic acid를 전처치한 후 KBrO₃를 투여하였을 때 FRTL-5 세포의 생존율은 80% 이상 농도에 의존적으로 증가하였으며, retinol과 α-tocopherol을 각각 전처치한 후 세포의 생존율은 70% 이상 증가하였고, EGCG를 전처치하였을 때 세포생존율은 60%대로 약간 증가하였다. 그러나 고농도에서 retinol과 EGCG의 전처치는 세포생존율을 오히려 감소시켰다. Melatonin, D-methionine을 전처치한 후 KBrO₃를 투여하였을 때 FRTL -5 세포의 생존율은 KBrO₃만을 처치하였을 때와 비교해서 변화가 없었다. 이상의 결과에서 FRTL-5 갑상선세포에서 KBrO₃에 의해 유발된 산화독성에 대하여 일상에서 섭취할 수 있는 녹차나 비타민 제제는 어느 정도 보호효과가 있을 것으로 생각되나 무분별하게 과다한 섭취는 오히려 산화독성을 유발할 가능성이 있어 항산화물질의 섭취에 주의를 기울여야 할 것이며, 이러한 항산화물질들의 복합적인 투여에 대한 연구가 추가적으로 진행되어야 할 것으로 생각된다. The present study was designed to evaluate the protective effects of antioxidative agents on cytotoxicity mediated by potassium bromate (KBrO₃), which is a prooxidant and carcinogen, in FRTL-5 rat thyroid cells. FRTL-5 cells were primarily incubated in the presence of 200 μM KBrO₃ for 4 hours to induce cytotoxicity and measured the cell viability using MTS-based cell proliferation assay. The protective effect of antioxidative agents, including N-acetylcysteine (NAC), L-2-oxothiazoline -4-carboxylate (OTC), L-N(G)-nitroarginine methylester (L-NAME), melatonin, D-methionine, minocycline, (-)-epigallocatechin-3-gallate (EGCG), retinol, ascorbic acid or α-tocopherol on KBrO₃-induced cytotoxicity in FRTL-5 cells was evaluated with pretreating each antioxidant for 2-24 hours prior to the exposure of KBrO₃. Treatment with 200 μM of KBrO₃ caused a reduction of cell viability to about 60% comparing to control. The reduction of cell viability by 200 μM KBrO₃ was blocked by pretreating 0.05-1 mM of NAC or OTC in a dose-dependent manner. Pretreatment with minocyline was also found to promote cell viability by KBrO₃, however, L-NAME has no effect on the cell viability. Furthermore, pre-treatment with ascorbic acid, retinol, α-tocopherol and EGCG showed some protective effect on FRTL-5 cells against KBrO₃ -induced cytotoxicity, respectively. However, retinol and EGCG in high concentration further decreased the cell viability less than that by 200 μM KBrO₃, respectively. On the other hand, L-NAME, melatonin or D-methionine has no protective effect against KBrO₃-induced cytotoxicity on FRTL-5 cells. In conclusion, these results demonstrate that the intake of ascorbic acid, retinol, α-tocopherol or EGCG may be benefit to protect the thyroid cells against KBrO₃-induced oxidative toxicity. However, it is recommended to have a caution when to intake these antioxidants particularly in high concentrations because of their prooxidant effects.
URI
http://kumel.medlib.dsmc.or.kr/handle/2015.oak/11778
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3. Thesis (학위논문) > 1. School of Medicine (의과대학) > 박사
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