N of SCD-1. (d) Quantification benefits on the CCR9 Antagonist Synonyms expression of SREBP1. Information were expressed as imply SD of 3 independent experiments (n = 3). ## p 0.01, compared with vehicle-treated manage cells (Con); p 0.05 and p 0.01, compared with OA-treated cells (OA).two.5. Kaempferol and Kaempferide Decreased Expression of PPAR and C/EBP in OA-Treated HepG2 Cells To figure out irrespective of whether kaempferol and kaempferide regulate adipogenesis, protein levels of PPAR and C/EBP, two adipogenic transcription aspects, have been determined by western blot. As shown in Figure 6, incubation with kaempferol (10 and 20 ) and kaempferide (5, 10 and 20 ), reduced the expression of PPAR in HepG2 cells (p 0.01), compared with OA group. A dose-dependent manner was observed for kaempferol. Moreover, incubation with kaempferol and kaempferide triggered a trend of reduction in expression of C/EBP, though statistical significance was not realized. These outcomes suggest kaempferol and kaempferide may well lessen lipid accumulation by decreasing the expression of adipogenesis-related proteins.Int. J. Mol. Sci. 2021, 22,8 ofFigure 6. Kaempferol and kaempferide inhibited expression of PPAR and C/EBP in OA-treated HepG2 cells. HepG2 cells have been incubated with different concentrations of kaempferol or kaempferide inside the presence of 0.5 mM OA for 48 h and followed by western blot analysis on the expression of PPAR and C/EBP. (a) Representative blots. (b) Quantification benefits of your expression of PPAR. (c) Quantification results in the expression of C/EBP. Data have been expressed as mean SD of three independent experiments (n = three). ## p 0.01, compared with vehicle-treated control cells (Con); p 0.05 and p 0.01, compared with OA-treated cells (OA).2.six. Kaempferol and Kaempferide Decreased Expression of Estrogen receptor Antagonist Compound Perilipin-1 and Caveolin-1 in OA-Treated HepG2 Cells Expression of lipid droplet-associated proteins, Perilipin-1 and Caveolin-1, was determined by western blot. As shown in Figure 7, compared with OA group, incubation with kaempferol (five, ten and 20 ) and kaempferide (five, ten and 20 ) decreased the expression of Perilipin-1 in HepG2 cells (p 0.01), having a dose-dependent effect becoming observed. Furthermore, kaempferol (10 and 20 ) and kaempferide (10 and 20 ) decreased the expression of Caveolin-1 (p 0.05, Figure 7). These final results recommend kaempferol and kaempferide may well lessen lipid accumulation by decreasing the expression of lipid droplet proteins.Int. J. Mol. Sci. 2021, 22,9 ofFigure 7. Kaempferol and kaempferide inhibited expression of Perilipin-1 and Caveolin-1 in OA-induced HepG2 cells. HepG2 cells were incubated with distinct concentrations of kaempferol or kaempferide in the presence of 0.five mM OA for 48 h followed by western blot evaluation on the expression of Perilipin-1 and Caveolin-1. (a) Representative blots. (b) Quantification results on the expression of Perilipin-1. (c) Quantification final results of your expression of Caveolin-1. Data had been expressed as imply SD of 3 independent experiments (n = 3). # p 0.05 and ## p 0.01, compared with vehicle-treated handle cells (Con); p 0.05 and p 0.01, compared with OA-treated cells (OA).2.7. Kaempferol and Kaempferide Reduced Expression of Nrf2 and HO-1 in OA-Treated HepG2 Cells Beneath oxidative tension situations, cells respond by activation of its antioxidant defense machinery to combat the cellular damage. We investigated the impact of kaempferol and kaempferide on oxidative stress by analyzing the adjustments inside the Nrf2/HO-1 pathway.