Effects of Valproic Acid on the Survival of Human Tennon's Capsule Fibroblasts

See Eun Lee; Jae Woo Kim

doi:10.3341/jkos.2018.59.11.1056

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Lee and Kim: Effects of Valproic Acid on the Survival of Human Tennon's Capsule Fibroblasts

Original Article

Journal of the Korean Ophthalmological Society 2018; 59(11): 1056-1061.

Published online: 16 November 2018

DOI: https://doi.org/10.3341/jkos.2018.59.11.1056

Effects of Valproic Acid on the Survival of Human Tennon's Capsule Fibroblasts

See Eun Lee, MD, Jae Woo Kim, MD, PhD

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

Address reprint requests to Jae Woo Kim, MD, PhD. Department of Ophthalmology, Daegu Catholic University Medical Center, #33 Duryugongwon-ro 17-gil, Nam-gu, Daegu 42472, Korea. Tel: 82-53-650-4728, Fax: 82-53-627-0133, jwkim@cu.ac.kr

Received 28 February 2018 Revised 21 March 2018 Accepted 18 October 2018

The Korean Ophthalmological Society

(open-access):

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

To investigate the effects of valproic acid on the survival of cultured human Tenon's capsule fibroblasts (HTFBs).

Methods

Primary cultured HTFBs were exposed to 0, 0.25, 0.5, and 1.0 mM valproic acid with or without 0, 1.0, 2.5 µg/mL mitomycin C, and incubated for 5 days. Cell survival was assessed using an MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide) assay and the degree of apoptosis was assessed by flow cytometry using annexin-V/propidium iodide double staining.

Results

Valproic acid decreased the survival of HTFBs in a dose-dependent manner, and survival was further decreased by adding mitomycin C to valproic acid. Both valproic acid and mitomycin C induced apoptosis of HTFBs. Valproic acid induced less apoptosis than mitomycin C.

Conclusions

Valproic acid decreased the cellular survival of HTFBs and induced apoptosis. The antiproliferative effects of valproic acid were further enhanced by the addition of mitomycin C.

Keywords: Apoptosis, Fibroblast, Flow cytometry, Mitomycin C, Valproic acid

Figures and Tables

Figure 1

Effect of valproic acid and mitomycin C on the survival of human Tenon's capsule fibroblasts. Both valproic acid (V, mM) and mitomycin C (M, µg/mL) decreased cellular survival in a dose-dependent manner (^*p < 0.05).

Figure 2

Additive effect of valproic acid (V, mM) on the survival of human Tenon's capsule fibroblasts when co-exposed to 1.0 µg/mL of mitomycin C (M). Valproic acid further decreased mitomycin C-induced reduction of cellular survival significantly (^*p < 0.05).

Figure 3

Additive effect of valproic acid (V, mM) on the survival of human Tenon's capsule fibroblasts when co-exposed to 2.5 µg/mL of mitomycin C (M). Valproic acid further decreased mitomycin C-induced reduction of cellular survival significantly (^*p < 0.05).

Figure 4

Flow cytometric analysis of apoptosis using annexin-propidium iodide (PI) double staining. Cells in quadrant B1, B2, B3, B4 represent necrotic cells, late apoptotic cells, living cells and early apoptotic cells, respectively. (A) Exposed to 0.5 mM valproic acid. (B) Exposed to 2.5 µg/mL of mitomycin C. Cell count = 10,000. FITC = fluorescein isothiocyanate.

Table 1

Results of flow cytometric analysis of apoptosis induced by valproic acid (VPA) and mitomycin C (MMC)

Values are presented as mean ± SD (%). ‘B1’ means ‘only propidium iodide (PI) stained (necrotic cells)’, ‘B2’ means ‘Annexin and PI stained (late apoptotic cells)’. ‘B3’ means ‘unstained (live cells)’, ‘B4’ means ‘only Annexin stained (early apoptotic cells)’.

Table 2

Additive effect of valproic acid (VPA) on the apoptosis of human Tenon's capsule fibroblasts when co-exposed to mitomycin C (MMC)

^*Represent % apoptosis as mean ± standard deviation.

Notes

^{Conflicts of Interest} The authors have no conflicts to disclose.

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Effects of Valproic Acid on the Survival of Human Tennon's Capsule Fibroblasts

Abstract

Purpose

Methods

Results

Conclusions

Figures and Tables

Figure 1

Effect of valproic acid and mitomycin C on the survival of human Tenon's capsule fibroblasts. Both valproic acid (V, mM) and mitomycin C (M, µg/mL) decreased cellular survival in a dose-dependent manner (*p < 0.05).

Figure 2

Additive effect of valproic acid (V, mM) on the survival of human Tenon's capsule fibroblasts when co-exposed to 1.0 µg/mL of mitomycin C (M). Valproic acid further decreased mitomycin C-induced reduction of cellular survival significantly (*p < 0.05).

Figure 3

Additive effect of valproic acid (V, mM) on the survival of human Tenon's capsule fibroblasts when co-exposed to 2.5 µg/mL of mitomycin C (M). Valproic acid further decreased mitomycin C-induced reduction of cellular survival significantly (*p < 0.05).

Figure 4

Table 1

Results of flow cytometric analysis of apoptosis induced by valproic acid (VPA) and mitomycin C (MMC)

Table 2

Additive effect of valproic acid (VPA) on the apoptosis of human Tenon's capsule fibroblasts when co-exposed to mitomycin C (MMC)

Notes

References

Effect of valproic acid and mitomycin C on the survival of human Tenon's capsule fibroblasts. Both valproic acid (V, mM) and mitomycin C (M, µg/mL) decreased cellular survival in a dose-dependent manner (^*p < 0.05).

Additive effect of valproic acid (V, mM) on the survival of human Tenon's capsule fibroblasts when co-exposed to 1.0 µg/mL of mitomycin C (M). Valproic acid further decreased mitomycin C-induced reduction of cellular survival significantly (^*p < 0.05).

Additive effect of valproic acid (V, mM) on the survival of human Tenon's capsule fibroblasts when co-exposed to 2.5 µg/mL of mitomycin C (M). Valproic acid further decreased mitomycin C-induced reduction of cellular survival significantly (^*p < 0.05).