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Role of Hydrogen Sulfide in the Survival of Fibroblasts and Fibroblast-mediated Contraction of Collagen Gel
Journal of the Korean Ophthalmological Society ; : 975-981, 2019.
Article in Korean | WPRIM | ID: wpr-766834
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.
Subject(s)

Full text: Available Index: WPRIM (Western Pacific) Main subject: RNA, Messenger / Polymerase Chain Reaction / Actins / Collagen / Cicatrix / Apoptosis / Tenon Capsule / Fibroblasts / Flow Cytometry / Gels Limits: Humans Language: Korean Journal: Journal of the Korean Ophthalmological Society Year: 2019 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: RNA, Messenger / Polymerase Chain Reaction / Actins / Collagen / Cicatrix / Apoptosis / Tenon Capsule / Fibroblasts / Flow Cytometry / Gels Limits: Humans Language: Korean Journal: Journal of the Korean Ophthalmological Society Year: 2019 Type: Article