Fig 2: Alteration of PKLR activity by pravastatin treatment in lean C57BL/6 mouse kidney and liver. Mouse kidney (A) and liver (B) were lysed by activity assay buffer and then pyruvate measured in 50 µg tissue. The generated pyruvate is oxidized by pyruvate oxidase, while producing light at 570 nm wavelength. **P < 0.01 vs. CTRL (n = 5).

Fig 3: Effects of pravastatin on glucose metabolism-related enzyme expression in HK-2 cells and HepG2 cells under high-cholesterol conditions. HK-2 cell (A) and HepG2 cell (B) were treated with 1, 2, or 4 µM pravastatin (prava) plus 25-hydroxy cholesterol (chol.) for 24 or 48 h. Total protein lysates were western blotted using the antibodies; PKLR (59 kD), PFK-1 (85 kD), PEPCK (62 kD), and G6PC (36 kD). ß-actin was used to confirm equal loading. (C) Bar graphs representing quantitative differences in expressions of PKLR. Results are means ± SEMs (n = 3). *P < 0.05, **P < 0.01 vs. the 48 h-untreated control (-/-, CTRL).

Fig 4: Effects of pravastatin on PKLR expression in high-fat diet-fed C57BL/6 mice. C57BL/6 mice were fed a high-fat diet with or without pravastatin (0.01%, w/w) for 16 weeks. (A) Kidney tissue lysates (40 µg) were loaded onto gels and immunoblotted; ß-actin was used to confirm equal loading. (B) Bar graphs representing quantitative differences in PKLR protein expression. Body weights (C) and blood glucose levels (D) were measured at 6, 10, and 22 weeks of age and 10 and 22 weeks of age, respectively. *P < 0.05; **P < 0.01 compared with untreated mice. Data are presented as mean ± SEM (CTRL; n = 5, Pravastatin; n = 9).

Fig 5: Effects of pravastatin on glucose metabolism-related enzymes in HK-2 cells and HepG2 cells under normal conditions. HK-2 cell (A) and HepG2 cell (B) were treated with 1, 2, or 4 µM pravastatin for 24, 48, or 72 h. Western blotting was used to evaluate the protein expression of PFK-1, PKLR, PEPCK, and G6PC in the treated cells. Cell lysates (40 µg) were loaded onto gels and immunoblotted; ß-actin was used to confirm equal loading. (C) Bar graphs representing quantitative differences in expressions of PKLR. Results are means ± SEMs (n = 3). *P < 0.05, **P < 0.01 vs. CTRL.