See Eun Lee; Keun Hae Kim; Jae Woo Kim

doi:10.3341/jkos.2015.56.8.1268

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Lee, Kim, and Kim: Flow Cytometric Analysis of the Effects of Resveratrol on the Survival of Human Tennon’s Capsule Fibroblasts

Original Article

J Korean Ophthalmol Soc 2015;56(8):1268-1273.

Published online: 29 August 2015

DOI: https://doi.org/10.3341/jkos.2015.56.8.1268

Flow Cytometric Analysis of the Effects of Resveratrol on the Survival of Human Tennon’s Capsule Fibroblasts

See Eun Lee, M.D., Keun Hae Kim, M.D., Jae Woo Kim, M.D., PhD

Department of Ophthalmology, Catholic University of Daegu School of Medicine, Daegu, Korea

Address reprint requests to Jae Woo Kim, MD, PhD Department of Ophthalmology, Daegu Catholic University Hospital, #33 Duryugongwon-ro 17-gil, Nam-gu, Daegu 705-718, Korea Tel: 82-53-650-4728, Fax: 82-53-627-0133 E-mail: jwkim@cu.ac.kr

Received 12 February 201530 March 2015 Accepted 19 June 2015

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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 μ M resveratrol for 3 days. Cellular survival was assessed us-ing 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 μ M ( p = 0.04). In flow cytometric analysis, 10 μ M re-sveratrol did not affect the degree of apoptosis ( p = 0.89), but 100 μ M resveratrol increased the degree of apoptosis ( p = 0.003). Both 10 and 100 μ M 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 an-tiproliferative effects on HTFBs.

Keywords: Apoptosis, Fibroblast, Flow cytometry, Resveratrol, Survival

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Figure 1.

Effects of resveratrol on the survival of human Tenon's capsule fibroblasts. Resveratrol decreased cellular survival significantly in a dose-dependent manner. ^* p < 0.05.

Figure 2.

Flow cytometric analysis of apoptosis using annexin-PI double staining. Cells in quadrant B1, B2, B3, B4 rep-resents necrotic cells, late apoptotic cells, living cells and early apoptotic cells, respectively. (A) Unstained control. (B) Exposed to 100 μ M resveratrol. FITC = fluorescein isothiocyanate; PI = propidium iodide.

Figure 3.

Flow cytometric analysis of apoptosis using annex-in-PI double staining. 100 μ M resveratrol increased the degree of apoptosis significantly compared to non-exposed control. PI = propidium iodide. ^* p < 0.05.

Figure 4.

Flow cytometric analysis of necrosis using annex-in-PI double staining. 10, 100 μ M resveratrol did not affect the degree of necrosis significantly compared to non-exposed con-trol ( p > 0.05). PI = propidium iodide.

Figure 5.

Effect of resveratrol on the production of nitric ox-ide in human Tenon's capsule fibroblasts. Resveratrol in-creased nitric oxide production significantly in a dose-depend-ent manner. ^* p < 0.05.

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