Table 1.
The body and fat weight, food intake and lipids in the obese agouti-mice and the effect of metformin treatment.
Table 2.
The plasma levels of glucose, insulin and leptin and the intrahypothalamic leptin and insulin content in obese agouti-mice and the effect of metformin treatment.
Fig 1.
Glucose curves and AUC0-120 in GTT in obese agouti-mice and the effect of metformin treatment.
(A) The glucose concentration curves in the glucose tolerance test. (B) The AUC0-120 values for the glucose concentration curves during the time interval from 0 to 120 min. The data are presented as the M ± SD. *—the difference between the control mice and all the studied groups of agouti-mice is significant at P < 0.05; #—the difference between the Group Ay/a and MF-treated agouti-mice is significant at P < 0.05. In each group: n = 5.
Fig 2.
The histological evidence of steatosis in the liver of obese agouti-mice, and the improving effect of long-term metformin treatment (600 mg/kg/day).
A, B–Group a/a (control); C, D–Group Ay/a; E, F–Group Ay/a-M600. The liver histology was evaluated using the staining of liver sections with hematoxylin and eosin (A, C, E), while the hepatic lipid deposition was evaluated by the Sudan III-staining (B, D, F). The orange inclusions and granules after the Sudan III staining are the lipid and lipoproteins accumulations located within hepatocytes. The liver sections from animals of each of the three studied groups were mounted on the same glass. Scale bars, 100 μm.
Fig 3.
The gene expression of the proteins involved in apoptosis and inflammation in the liver of obese agouti-mice and the effect of metformin treatment.
The levels of mRNA expression of the Bax, Bcl-2, IL1beta and TNFalpha genes encoding pro-apoptotic factor Bax, anti-apoptotic factor Bcl-2, the inflammatory cytokines interleukin 1β and TNFα are normalized by the expression of the reference 18s rRNA gene. The relative mRNA expression is calculated with the respect to control group. The data are presented as the M ± SEM, n = 5. *—the difference between the Groups a/a and Ay/a-M600 is significant at the P < 0.05, #—the difference between the Groups Ay/a and Ay/a-M600 is significant at the P < 0.05.
Fig 4.
Phosphorylation of AMPK α-subunits in the hypothalamus of obese agouti-mice and the effect of metformin treatment.
(A) The histograms for the ratio of the Thr172- and Ser485/491-phosphorylated α1/2-AMPK and the non-phoshorylated α2-AMPK. (B) and (C) The Western blotting for the Thr172- and Ser485/491-phosphorylated forms of α1/2-AMPK, respectively. The data are presented as the M ± SEM. *—the difference between the Groups a/a and Ay/a is significant at the P < 0.05, #—the difference between the Groups Ay/a and Ay/a-M600 is significant at the P < 0.05. In each group: n = 5.
Fig 5.
Phosphorylation of Akt-kinase in the hypothalamus of obese agouti-mice and the effect of metformin treatment.
(A) The histograms for the ratio of the Ser473- and Thr308-phosphorylated and non-phosphorylated Akt-kinase. (B) and (C) The Western blotting for the Ser473- and Thr308-phosphorylated forms of Akt kinase, respectively. The data are presented as the M ± SEM. #—the difference between the Groups Ay/a and Ay/a-M600 is significant at the P < 0.05. In each group: n = 5.
Fig 6.
Phosphorylation of STAT3 in the hypothalamus of obese agouti-mice and the effect of metformin treatment.
(A) The histograms for the ratio of the Tyr705-phosphorylated and non-phosphorylated STAT3. (B) The Western blotting for the Tyr705-phosphorylated STAT3. The data are presented as the M ± SEM. *—the difference between the Groups a/a and Ay/a is significant at the P < 0.05. In each group: n = 5.
Fig 7.
The expression of the genes in the hypothalamus of obese agouti-mice and the effect of metformin treatment.
The levels of mRNA expression of the Lepr, Pomc, Agrp, Npy, Mc3r and Mc4r genes encoding LepR, POMC, AgRP, NPY, MC3R and MC4R are normalized by the expression of the reference 18s rRNA and Hprt genes. The relative mRNA expression is calculated with the respect to control group. The data are presented as the M ± SEM, n = 5. *—the difference between the Groups a/a and Ay/a-M600 is significant at the P < 0.05, #—the difference between the Groups Ay/a and Ay/a-M600 is significant at the P < 0.05.
Fig 8.
Metabolic and hormonal parameters and hypothalamic regulation in obese agouti-mice and the effect of metformin treatment.