Effect of Alternate day fasting over Metabolic dysfunction-associated steatohepatitis in adult offspring of dams exposed to cafeteria diet during pregnancy and lactation.

Introductions and Objectives

Metabolic dysfunction-associated steatohepatitis (MASH) is a liver disease characterized by lipid accumulation and inflammation that can be exacerbated by cafeteria diets (CAF) exposition during pregnancy and lactation, whereas Alternate day fasting (ADF) improves metabolic parameters. Evaluate the effect of ADF and CAF maternal programming on MASH-associated markers in the offspring.

Materials and Patients

To assess the effect of maternal programming, we elaborated a mice model using 8-week C57BL6 females exposed to a CAF (cafeteria) diet (39% carbs, 49% fats, 12% proteins and sodium 231.8 mg) during 3 weeks of mating, 3 weeks of gestation and 3 weeks of lactation. For maternal programming control, we fed females with a Chow or control diet (57% carbs, 13% fats, 30% proteins and sodium 105 mg) during 3 weeks of mating, 3 weeks of gestation and 3 weeks of lactation. After weaning, the offspring were fed a control diet until they were 8 weeks old. They were then divided into four groups (Control n=8, Control + ADF n=8, CAF n=8, CAF+ ADF n=8) and an alternate day Fasting (ADF) protocol was initiated for 5 weeks. At the end of the fasting protocol, plasma samples were taken and beta-hydroxybutyrate (BHB) concentration was measured; in addition, samples of the left lateral lobe of the liver were taken at slaughter to evaluate by qPCR the effect of intermittent fasting on the expression of metabolic function markers involved during MASH: fibrosis (TGFβ, Col1a1), steatosis (PLIN2, ApoB100, Mylcd, PPARPα) and inflammation (Mcp-1).

Results

Groups treated with ADF showed an increase in plasma BHB concentration of 400 μmol compared to non-fasted groups. However, no significant difference was found between the control +ADF and CAF + ADF groups, so no effect of maternal programming with CAF diet on BHB production was observed. Additionally, the relative expression of mRNA from fibrosis-associated markers such as Col1a1 showed an 84% decrease in the CAF maternal programming model, 80% in the Control + ADF group and 88% in the CAF + ADF model with respect to control. Levels of mRNA-Plin2, involved in lipid droplet formation, decreased by 57% in the CAF group, 48% in Control +ADF and 79% in CAF+ADF. On the other hand, mRNA-Mcp-1 levels (chemokine) showed a decrease of 14.36% in CAF, 46.42% in Control + ADF and 62.68% in CAF+ ADF with respect to control.

Conclusions

The model of alternate-day fasting (ADF) showed an increased plasma BHB, but we did not observe a maternal programming effect on the concentration of betahydroxybutyrate. Interestingly, maternal programming and ADF reduce the expression of MASH-associated markers involved in fibrosis, lipid droplet formation and inflammation in this mouse model.

Ethical statements: This project was authorized by the ethics committee of the Universidad Autónoma de Nuevo León with the registration number B120-00,004.

Declaration of interest: None.

Funding: This work was supported by the National Council of humanities, Science and Technology in Mexico (CONACHYT) for Martín García-Juarez, (Grant number: 631654).