Suggesting kaempferol and CYP2 Inhibitor Species kaempferide usually do not affect cell viability of OA-treated HepG2 cells.Int. J. Mol. Sci. 2021, 22,5 ofFigure three. Modifications in viability of HepG2 cells just after incubation with kaempferol and kaempferide. (a) Chemical structure of kaempferol. (b) Chemical structure of kaempferide. (c) HepG2 cell viability after incubation with kaempferol. (d) HepG2 cell viability soon after incubation with kaempferide. (e) No alter in HepG2 cell viability by co-incubation of OA and kaempferol for 48 h. (f) No alter in HepG2 cell viability by co-incubation of OA and kaempferide for 48 h. Data had been expressed as Mean SD of 3 independent experiments (n = 3). p 0.01, compared with vehicle-treated handle.two.three. Kaempferol and Kaempferide IL-4 Inhibitor medchemexpress suppressed Lipid accumulation in OA-Treated HepG2 Cells To investigate whether kaempferol and kaempferide influence intracellular lipid accumulation, oil red O staining was performed. 0.5 mM OA brought on prominent raise lipid droplets accumulation in HepG2 cells, compared together with the control group (Figure 4a,b). Noticeably, incubation with kaempferol and kaempferide for 48 h reduced the accumulation of intracellular lipid droplets in a dose-dependent manner, compared with OA group. In addition, kaempferide lowered the intracellular TG levels at concentration of 10 and 20 (p 0.01), compared with all the OA group (Figure 4c). Kaempferol treatment induced a trend of reduction in TG content material, but statistical significance was not accomplished. The results recommend that kaempferol and kaempferide attenuate OA-induced lipid accumulation in HepG2 cells.Int. J. Mol. Sci. 2021, 22,six ofFigure four. Kaempferol and kaempferide suppressed lipid accumulation in OA-induced HepG2 cells. HepG2 cells had been incubated with distinct concentrations of kaempferol or kaempferide in the presence of 0.5 mM OA for 48 h. (a) Oil red O staining inside the cultured HepG2 cells. (b) Visualization of intracellular lipid droplets in HepG2 cells under microscope (100magnification). (c) Quantification of intracellular TG contents in HepG2 cells. Data had been expressed as imply SD of 3 independent experiments (n = 3). ## p 0.01, compared with vehicle-treated handle cells (Con); p 0.01, compared with OA-treated cells (OA).2.4. Kaempferol and Kaempferide Decreased Expression of SREBP1, FAS and SCD-1 in OA-Treated HepG2 Cells To ascertain the underlying mechanism for the inhibitory effect of kaempferol and kaempferide on lipid accumulation, expression of lipogenesis-related proteins, SREBP1, FAS and SCD-1 had been analyzed by western blot. As shown in Figure five, kaempferide dosedependently decreased the expression of SREBP1 in HepG2 cells (p 0.01), compared with OA group. Reduction was also observed in expression of FAS and SCD-1 (p 0.01), which was regulated by SREBP1. In contrast, therapy with kaempferol showed tiny impact on expression of SREBP1, FAS and SCD-1 (Figure 5). These findings recommend kaempferide could reduce lipid accumulation in OA-treated HepG2 cells through decreasing the expression of lipogenic proteins.Int. J. Mol. Sci. 2021, 22,7 ofFigure 5. Kaempferol and kaempferide reduced expression of SREBP1, FAS and SCD-1 in OA-treated HepG2 cells. HepG2 cells had been treated with various concentrations of kaempferol or kaempferide within the presence of 0.5 mM OA for 48 h followed by western blot analysis of expression of SREBP1, FAS and SCD-1. (a) Representative blots. (b) Quantification benefits in the expression of FAS. (c) Quantification final results in the expressio.
