Research Paper Volume 13, Issue 23 pp 25342—25364

Protective effects of galangin against H2O2/UVB-induced dermal fibroblast collagen degradation via hsa-microRNA-4535-mediated TGFβ/Smad signaling

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Figure 3. Galangin attenuates UVB/H2O2-induced intracellular and mitochondrial ROS production as well as MMP imbalance in HS68 cells. HS68 cells were exposed to H2O2 (200 μM) for 1 h and then co-treated with galangin (30 μM) for 23 h. HS68 cells were exposed to UVB radiation (40 J/cm2) and then co-treated with galangin (30 μM) for 24 h. (A, B) Intracellular and mitochondrial ROS levels were determined by MitoSOX™ red mitochondrial superoxide indicator (2 μM) and 2′,7′-dichlorofluorescin diacetate (DCF-DA) (5 μM). The mitochondria positive for ROS fluoresced red, while green fluorescence indicated intracellular ROS levels. (C) Mitochondrial membrane potential (MMP) was determined by JC-1 staining. Red fluorescence indicated JC-1 dimers (intact mitochondria) and green fluorescence indicated JC-1 monomers (impaired mitochondria). The images were taken using a florescence microscope. Values are shown as mean ± SE. Quantification of the results is shown (n = 3) **P < 0.01, ***P < 0.001 vs. untreated control cells; #P < 0.05, vs. H2O2 or UVB-treated cells.