Fish Scale Collagen Peptides Protect against CoCl₂/TNF-α-Induced Cytotoxicity and Inflammation via Inhibition of ROS, MAPK, and NF-κB Pathways in HaCaT Cells

Figure 2

Inhibitory effects of FSCP on CoCl₂/TNF-α-induced ROS generation in HaCaT cells. Intracellular ROS levels were determined via fluorescence microscopy (a, c) or flow cytometry (b, d) using DCFH-DA. The increased ROS levels induced by 100 μM CoCl₂ treatment was significantly attenuated by treatment with 1 mg/mL FSCP for 24 h or NAC for 2 h (a, b). Elevated ROS levels were induced by treatment with 20 ng/mL TNF-α but were reversed by treatment with 1 mg/mL FSCP for 12 or 24 h (c, d). Staining intensities were measured using ImageJ software (a, c). Blue histograms represent ROS negative cells, red histograms represent ROS positive cells, and the median fluorescence intensity (MFI) of ROS expression is plotted along the y-axes (b, d). Results are presented as the means ± SD of three independent experiments. ^# versus the control and and versus the CoCl₂/TNF-α-treated group.

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