J Korean Ophthalmol Soc.  2015 Aug;56(8):1268-1273. 10.3341/jkos.2015.56.8.1268.

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

• Lee SE
• Kim KH
• Kim JW

• KMID: 2215081
• DOI: http://doi.org/10.3341/jkos.2015.56.8.1268

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 microM 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 microM (p = 0.04). In flow cytometric analysis, 10 microM resveratrol did not affect the degree of apoptosis (p = 0.89), but 100 microM resveratrol increased the degree of apoptosis (p = 0.003). Both 10 and 100 microM 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.

Keyword

Apoptosis; Fibroblast; Flow cytometry; Resveratrol; Survival

MeSH Terms

Apoptosis
Fibroblasts*
Flow Cytometry
Humans*
Necrosis
Tenon Capsule

Figure

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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.

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  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.

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  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.

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  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.

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  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.

Reference

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