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. 2022 Mar 6;18(5):2202–2219. doi: 10.7150/ijbs.65973

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Melatonin restores the TNF-α-induced metabolic disturbance of NP cells in vitro. (A and B) The expression of TNF-α in NP tissues from human and rat were detected by IHC. Representative images of different magnifications were displayed. Scale bars: 100 μm and 50 μm for 200× and 400× images, respectively. (C) RT-qPCR analysis was conducted to assess the mRNA expression of COL2A1 and MMP13 in human NP cells with the addition of vehicle(ethanol), TNF-α (10 ng/ml) and a gradient concentration of melatonin for 48 hours to determine the optimum concentration. After indicated treatment for 48 hours, RT-qPCR (D) and WB (E) analyses were conducted to assess the expression of COL2A1, ACAN, MMP13, and ADATMS5 in human NP cells. RT-qPCR (F) analysis was conducted to assess the expression of IL-1β and TNF-α. * means P < 0.05 compared with the vehicle group. # means P < 0.05 compared with the TNF-α group. Ns means not significant.