J Korean Ophthalmol Soc > Volume 60(10); 2019 > Article
Journal of the Korean Ophthalmological Society 2019;60(10):975-981.
DOI: https://doi.org/10.3341/jkos.2019.60.10.975    Published online October 15, 2019.
Role of Hydrogen Sulfide in the Survival of Fibroblasts and Fibroblast-mediated Contraction of Collagen Gel.
Hyeon Jin Park, Jae Woo Kim
Department of Ophthalmology, Daegu Catholic University School of Medicine, Daegu, Korea. jwkim@cu.ac.kr
황화수소가 섬유아세포의 생존과 교원질 겔의 수축에 미치는 영향
박현진 · 김재우
대구가톨릭대학교 의과대학 안과학교실
Correspondence:  Jae Woo Kim,
Email: jwkim@cu.ac.kr
Received: 24 January 2019   • Revised: 26 February 2019   • Accepted: 19 September 2019
Abstract
PURPOSE
To investigate the role of hydrogen sulfide in the survival and collagen gel contraction of cultured human Tenon's capsule fibroblasts (HTCFs). METHODS: Primarily cultured HTCFs were exposed to 0, 100, 200, or 300 µM hydrogen sulfide (sodium hydrogen sulfide, NaHS) for 2 days. Cellular survival was assessed by MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide) assay. Degree of apoptosis was assessed with flow cytometry using annexin-V/propidium iodide double staining. To evaluate the effect of NaHS on cellular transdifferentiation, HTCFs were stimulated with 5 ng/mL TGF-β1 and the level of expression of α-smooth muscle actin (SMA) mRNA was assessed using reverse-transcription polymerase chain reaction. The cells were embedded in collagen gel, and the amount of gel contraction was measured. RESULTS: NaHS at 300 µM reduced HTCF survival (p = 0.013); NaHS at both 200 and 300 µM increased apoptosis in a dose-dependent manner (p = 0.013 and p = 0.016). TGF-β1 increased the expression of α-SMA mRNA (p = 0.041); co-treatment with 100 µM NaHS decreased TGF-β1-induced α-SMA mRNA expression (p = 0.039) and inhibited collagen gel contraction. CONCLUSIONS: NaHS at high concentration reduced cellular survival and increased HTCF apoptosis. NaHS decreased TGF-β 1-induced increases in α-SMA mRNA expression and collagen gel contraction. Thus, hydrogen sulfide may suppress scar formation by inhibiting HTCF transdifferentiation and contraction of collagen gels.
Key Words: Apoptosis;Hydrogen sulfide;Survival;Transdifferentiation;Tenon capsule fibroblasts


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