유세포분석을 이용한 레즈브라트롤이 인체 테논낭 섬유아세포의 생존에 미치는 영향에 대한 연구
Flow Cytometric Analysis of the Effects of Resveratrol on the Survival of Human Tennon’s Capsule Fibroblasts
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목적: 레즈브라트롤은 세포의 종류에 따라 세포보호 효과를 나타내거나 독성을 나타낸다. 레즈브라트롤이 배양된 인체 테논낭 섬유아세포의 생존에 미치는 영향에 대해 알아보고자 하였다. 대상과 방법: 일차배양한 섬유아세포에 0, 10, 100 μM의 레즈브라트롤에 3일간 노출시킨 후 MTT assay를 이용하여 세포의 생존을측정하였고, annexin-V/propidium iodide 이중염색 후 유세포분석을 시행하여 세포고사에 미치는 영향을 측정하였다. 결과: 레즈브라트롤은 10 μM의 농도에서부터 섬유아세포의 생존을 유의하게 저하시켰다(p=0.04). 유세포분석에서 10 μM의 농도에서는 섬유아세포의 세포고사를 유의하게 유발하지 않았으나(p=0.89), 100 μM의 농도에서는 섬유아세포의 세포고사를 유의하게 증가시켰다(p=0.003). 10 μM와 100 μM의 농도에서 세포괴사에는 유의한 영향을 미치지 않았다(p=0.74, 0.33). 결론: 배양된 인체의 테논낭섬유아세포에 대해 레즈브라트롤은 세포의 생존을 저하시키며 세포고사를 유발하므로 인체의 테논낭섬유아세포에 대해 레즈브라트롤은 항증식작용을 나타낼 수 있을 것이다.
Abstract
Purpose: Resveratrol exerts cytoprotective or cytotoxic effects according to cell type. This study was performed to evaluate the effects of resveratrol on the survival of cultured human Tenon’s capsule fibroblasts (HTFBs). Methods: Primarily cultured HTFBs were exposed to 0, 10, or 100 μM resveratrol for 3 days. Cellular survival was assessed using the MTT assay and degree of apoptosis was analyzed with flow cytometry using annexin-V/propidium iodide double staining. Results: Resveratrol decreased the survival of HTFBs after exposure to 10 μM (p = 0.04). In flow cytometric analysis, 10 μM resveratrol did not affect the degree of apoptosis (p = 0.89), but 100 μM resveratrol increased the degree of apoptosis (p = 0.003). Both 10 and 100 μM resveratrol did not affect the degree of necrosis (p = 0.74, 0.33). Conclusions: Resveratrol decreased cellular survival of cultured HTFBs and induced apoptosis. Thus, resveratrol may exert antiproliferative effects on HTFBs.
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