Melatonin increases collagen content accumulation and Fibroblast Growth Factor-2 secretion in cultured human cardiac fibroblasts

Figures

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  Fig. 1. Total cell number counted in Bürker chamber (A), necrotic cell count recognized by trypan blue elimination assay (B), and fibroblasts proliferation measured by BrdU method (C) in controls (CTR) DMSO-treated cultures (DMSO) and fibroblasts administered with melatonin at the concentrations of 1 µM (MLT-1) and 0.1 µM (MLT-0.1). In A and B, values represent medians ± interquartile ranges (n = 7–8). Kruskal–Wallis’s test followed by Mann–Whitney U test; In C, each value represents mean ± SD. One-way ANOVA followed by Shaffer’s post hoc test

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  Fig. 2. Total cell number (A), necrotic cell count (propidium iodide fibroblasts; (B) early (C) and late (D) apoptotic cell number in controls (CTR), DMSO-treated cultures (DMSO) and fibroblasts administered with melatonin at the concentrations 0.1 µM (MLT-0.1). All measurements were performed using an Annexin-FITC Apoptosis detection kit. Values represent medians ± interquartile ranges (n = 6–7). Kruskal–Wallis’s test followed by Mann–Whitney U test

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  Fig. 3. Intracellular collagen (A), extracellular collagen (B), and total collagen (C) content within controls (CTR), DMSO-treated cultures (DMSO), and fibroblasts administered with melatonin at the concentration of 1 µM (MLT-1) and 0.1 µM (MLT-0.1). Collagen was evaluated by the Woessner method. Col1A1 (D) and Col3A1 (E) gene relative expression were measured by real-time PCR in controls (CTR) DMSO-treated cultures (DMSO) and fibroblasts administered with melatonin at the concentration of 0.1 µM (MLT-0.1). Values represent medians ± interquartile ranges; n = 7–8 (A, B, C); n = 3–5 (D, E). Kruskal–Wallis’s test followed by Mann–Whitney U test

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  Fig. 4. Glycosaminoglycan content (A) evaluated by 1,9-dimethylmethylene blue assay, within controls (CTR) DMSO-treated cultures (DMSO) and fibroblasts administered with melatonin at the concentration of 0.1 µM (MLT-0.1). FGF-2 (B), cardiotrophin (C), and MMP-2 (D) content evaluated by ELISA, in controls (CTR), DMSO-treated cultures (DMSO), and fibroblasts administered with melatonin at the concentration of 0.1 µM (MLT-0.1). Values represent medians ± interquartile ranges (n = 7–10). Kruskal–Wallis’s test followed by Mann–Whitney U test