Nopal and Pirfenidone Ameliorate Epididymal Fat Weight and Anthropometric Parameters in a Mouse Obesity Model with Diethylnitrosamine

Abstract

Introduction and Objectives

Obesity is associated with liver diseases. Mexico has a high prevalence of obesity and metabolic dysfunction-associated steatotic liver disease (MASLD). Nopal and Pirfenidone (PFD) increase insulin signaling and decrease hepatic steatosis in obese mice. We investigated PFD and nopal on anthropometric parameters in obese mice and with diethylnitrosamine (DEN).

Materials and Methods

Five-six-week-old male C57BL/6J mice were treated with a single dose of DEN (25 mg/kg) and fed with a high fat diet (HFD, 60 kcal% from fat: D12492) for 16 weeks. Animals were provided ad libitum access to food and water. The mice were randomly divided into eight groups (n=5 for each group): normal diet (ND), ND plus DEN (ND+DEN), HFD, HFD plus DEN (HFD+DEN), HFD plus DEN plus supplements (cellulose, maltodextrin, and casein; HFD+DEN+SUPPL), HFD plus DEN plus nopal (HFD+DEN+NOP), HFD plus DEN plus PFD (HFD+DEN+PFD), and HFD plus DEN plus NOP plus PFD (HFD+DEN+NOP+PFD). Freeze-dried nopal in fine powder (7%) were mixed with HFD and PFD (300 mg/kg/day) also were mixed with HFD. PFD dosage was adjusted according to body weight and mixed with the diets three times a week. Food intake was measured three times a week, and measurement of body weight each week. Experiments were done according to ARRIVE guidelines. Statistical significance of anthropometric data was determined for parametric data with one-way ANOVA analysis of variance followed by Tukey's post hoc analysis, statistical analyses were performed using SPSS.

Results

All HFD mice developed obesity (P≤0.05), and PFD and NOP plus PFD reduced body weight (P≤0.05). Liver weight was increased in HFD, HFD+DEN, and HFD+DEN+SUPPL groups (P≤0.05), and epididymal fat was increased in all HFD mice (P≤0.001), but NOP, PFD, and NOP plus PFD reduced liver weight (P≤0.05) and PFD, and NOP plus PFD decreased epididymal fat (P≤0.05). Heart weight was increased in HFD, HFD+DEN, HFD+DEN+SUPPL, and HFD+DEN+NOP groups (P≤0.05), but NOP (p=0.06), PFD, and NOP plus PFD reduced it (P≤0.001). The heart weight/body weight ratio was reduced in all mice with HFD (P≤0.001), and only NOP plus PFD increased the heart weight/body weight ratio (P≤0.05). Liver weight/body weight ratio tended to increase in HFD and HFD plus DEN, but decreased with NOP, PFD, and NOP plus PFD (P≤0.05). Body weight/tibia length ratio was increased in all HFD mice (P≤0.01) and decreased with PFD and NOP plus PFD (P≤0.05). Heart weight/tibia length ratio was increased in HFD, HFD+DEN, HFD+DEN+SUPPL, and HFD+DEN+NOP (P≤0.01), but decreased with PFD and NOP plus PFD (P≤0.001). Epididymal fat weight/tibia length ratio was increased in all HFD mice (P≤0.001) but decreased with PFD (P=0.07) and NOP plus PFD (P≤0.05) (Figure 1).

Conclusions

In this study, we showed that intervention with nopal and pirfenidone improved epididymal fat weight and anthropometrical parameters in obese mice with DEN, this effects observed are possibly due to increased insulin sensitivity and decreased hepatic steatosis by nopal and pirfenidone.