Objective. To observe the impact of quercetin and isoquercitrin on gluconeogenesis in hepatocytes. Methods. Mouse primary hepatocytes were cultured with lactic acid and pyruvic acid. After treatment with quercetin and isoquercitrin for 24 hours, the glucose concentration in the culture supernatant was determined. RT-PCR was used to detect the mRNAs of PEPCK, G6Pase, LKB1, and AMPKα. Protein levels of LKB1, AMPKα, and Thr172 phosphorylation were evaluated by Western blot. Results. The glucose concentration in the gluconeogenesis group (GN) was significantly higher than in the control group (C), but the glucose concentrations in the high level quercetin(group 80Q) and high level isoquercitrin (group 80I) were significantly lower than in the group GN, P<0.01. In the group 80Q, and group 80I, the mRNA levels of PEPCK and LKB1were significantly lower than in the group GN (P<0.01), and the G6Pase mRNA were significantly lower than in the group GN (P<0.05). The protein levels of LKB1 and the phosphorylation of AMPKα Thr172 in the group 80Q, group 40I, and group 80I were higher than in the group GN. The effects of quercetin and isoquercitrin on LKB1 and AMPKα were similar to those of metformin. Conclusions. Quercetin and isoquercitrin inhibit gluconeogenesis in hepatocytes, which may be related to the LKB1 upregulation and phosphorylation of AMPKα.
ABSTRACT. We examined the influence of caffeine on the proliferation and apoptosis of b cells cultured in vitro in the presence of the free fatty acid palmitic acid (PA). Different concentrations of caffeine (1-100 mM) and free fatty PA were added to cultured b cells. The MTT assay was used to analyze cell proliferative activity; flow cytometry was used to measure apoptosis and calculate the apoptosis rate. Compared with the blank control group, cells cultured with 500 mM PA for 24, 48, 72, and 96 h showed inhibition of pancreatic b cell proliferative activity. In the 10 and 25 mM caffeine groups cultured for 48, 72, and 96 h, b cell proliferative activity was much higher than that in the 500 mM PA group. The apoptosis rate in the 500 mM PA group was 40.55 ± 20.33%, which was higher than that in the blank control group. The apoptosis rates in the 10 and 25 mM caffeine group and the PA group were 19.12 ± 10.56 and 20.97 ± 9.75%, respectively, which was lower than that in the 500 mM PA group. At some concentrations, caffeine can improve free fatty PA levels and guide pancreatic b cell proliferation inhibition and cell apoptosis.
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