Journal of Endocrinology and Metabolism, ISSN 1923-2861 print, 1923-287X online, Open Access
Article copyright, the authors; Journal compilation copyright, J Endocrinol Metab and Elmer Press Inc
Journal website https://www.jofem.org

Original Article

Volume 12, Number 6, December 2022, pages 168-177

Combination of a Glucagon-Like Peptide 1 Analog and a Sodium-Glucose Cotransporter 2 Inhibitor Improves Lipid Metabolism Compared to the Monotherapies in Experimental Metabolic Syndrome

Figures

Figure 1.
Figure 1. Effects of liraglutide and canagliflozin on body mass and food intake of high-fat diet-fed rats. (a) Experimental design. (b) Body mass evolution, #P < 0.001 HFD vs. control, HFD-L, and HFD-LC. *P < 0.001 HFD-L vs. HFD-C. @P < 0.01 HFD-C vs. HFD, HFD-L, and HFD-LC. (c) Food intake evolution, expressed as the amount consumed per rat per day. *P < 0.05 HFD vs. HFD-LC. Values are means and SEM (n = 10 per experimental group). Control: rats on a regular diet; HFD: high-fat diet; HFD-L: HFD treated with liraglutide; HFD-C: HFD treated with canagliflozin; HFD-LC: HFD treated with liraglutide plus canagliflozin; SEM: standard error of the mean.
Figure 2.
Figure 2. Effects of liraglutide and canagliflozin on the glucose metabolism of high-fat diet-fed rats. (a) Oral glucose tolerance test (OGTT) performed after the treatment period. (b) Area under the curve (AUC) of the OGTT graph. ***P < 0.001 HFD vs. control, HFD-C, and HFD-LC. **P < 0.01 HFD vs. HFD-C and HFD-LC. ##P < 0.001 HFD-L vs. HFD. @@P < 0.001 HFD-C and HFD-LC vs. HFD. @P < 0.01 HFD-C and HFD-LC vs. HFD. #P < 0.05 HFD-L vs. HFD-C and HFD-LC. (c) Fasting blood glucose before OGTT. Values are means and SEM (n = 10 per experimental group). Control: rats on a regular diet; HFD: high-fat diet; HFD-L: HFD treated with liraglutide; HFD-C: HFD treated with canagliflozin; HFD-LC: HFD treated with liraglutide plus canagliflozin; SEM: standard error of the mean.
Figure 3.
Figure 3. Treatments with liraglutide and canagliflozin reduced liver fat of high-fat diet-fed rats. Liver slices were stained with oil red O to enable observation of hepatic steatosis (red). (a) Rats on a regular diet (control). (b) High-fat diet (HFD). (c) HFD + liraglutide (HFD-L). (d) HFD + canagliflozin (HFD-C). (e) HFD + canagliflozin + liraglutide (HFD-LC). (f) Morphometric analysis of liver steatosis after treatment period. (g) Liver weights after treatment period. Values are means and SEM. *P < 0.05, ****P < 0.0001. Scale bar = 50 µm. SEM: standard error of the mean; ns: non-significant.
Figure 4.
Figure 4. Effects of liraglutide and canagliflozin in the epididymal adipose tissue of high-fat diet-fed rats. Histology of the epididymal adipose tissue of: (a) rats on a regular diet (control), (b) high-fat diet (HFD), (c) HFD + liraglutide (HFD-L), (d) HFD + canagliflozin (HFD-C), (e) HFD + canagliflozin + liraglutide HFD-LC. (f) Morphometric analysis of adipocyte area after treatment period. (g) Epididymal weight after treatment period. ****P < 0.0001, **P < 0.001, *P < 0.05. Scale bar = 100 µm.

Tables

Table 1. Composition of the Experimental High-Fat Diet
 
Ingredient (g/kg)ControlHFD
The diet was formulated to meet the American Institute of Nutrition AIN-93G recommendations for rodent diets by Prag Solucoes. Control group was fed with a diet containing only soybean oil. The high fat diets containing soybean oil and lard. BHT: butylhydroxytoluene.
Casein140175
Cornstarch465.7192.6
Sucrose155155
Soybean oil100100
Lard-238
Fiber5050
AIN-93G mineral mix3535
AIN-93 vitamin mix1010
L-cystine1.81.8
Choline bitartrate2.52.5
BHT0.0080.06
Energy (kcal/g)15,90920,934
Protein (% of energy)7636
Carbohydrate (% of energy)1414
Fat (% of energy)1050

 

Table 2. Effects of Liraglutide and Canagliflozin in Metabolic Parameters of HFD Rats
 
Variables (mg/dL)ControlHFDHFD-LHFD-CHFD-LC
Values are expressed as means and SEM of six to eight animals per group. Total cholesterol: aP < 0.001 vs. control; bP < 0.05 vs. HFD; cP < 0.001 vs. HFD. Triglycerides: aP < 0.001 vs. control; bP < 0.001 vs. HFD; cP < 0.01 vs. HFD-LC and vs. HFD. HDL cholesterol: aP < 0.01 vs. control; bP < 0.01 vs. HFD; cP < 0.05 vs. HFD. LDL cholesterol: aP < 0.01 vs. control; bP < 0.01 vs. HFD; cP < 0.001 vs. HFD. Glucose urine: aP < 0.001 vs. control. HDL: high-density lipoprotein; LDL: low-density lipoprotein; AST: aspartate aminotransferase; ALT: alanine aminotransferase; HFD: high-fat diet; HFD-L: HFD treated with liraglutide; HFD-C: HFD treated with canagliflozin; HFD-LC: HFD treated with liraglutide plus canagliflozin; SEM: standard error of the mean.
Total cholesterol60.1 ± 4.8110.6 ± 14.5a79.5 ± 4.3b85.7 ± 5.8b63.3 ± 3.4c
Triglycerides82.9 ± 5.9215.9 ± 16.1a94.4 ± 9.3b138.9 ± 11.5c70.3 ± 10.3b
LDL cholesterol44.5 ± 8.092.5 ± 3.4a47.0 ± 11.0b58.2 ± 10.1b33.3 ± 5.5c
HDL cholesterol93.5 ± 7.938.7 ± 5.2a72.3 ± 8.6b64.1 ± 5.1c91.5 ± 8.9b
AST37.1 ± 3.636.0 ± 3.435.0 ± 4.533.0 ± 4.937.0 ± 5.3
ALT47.7 ± 5.946.5 ± 1.538.3 ± 2.153.0 ± 6.454.2 ± 2.4
Glucose in urine65.7 ± 3.368.0 ± 3.065.7 ± 3.1185.3 ± 2.8a184.7 ± 4.1a

 

Table 3. Summary of Effects on Metabolic Syndrome Parameters
 
Treatments
LiraCanaLira + Cana
Lira: liraglutide; Cana: canagliflozin; HDL: high-density lipoprotein.
Metabolic syndrome
  Body mass+++++
  Food intake+++++
  Fasting blood glucose+++++
  Abdominal fat+++++
  Liver fat++++++
  Dyslipidemia++++++
  Triglycerides++++++
  HDL cholesterol++++++