Effects Of High Fructose Exposure In Gestational And Lactating Rats On The Regulation Of Appetite By The PI3K/Akt/FoxO1 Pathway In The Hypothalamus Of The Offspring And Analysis Of Differentially Expressed Genes

Objective: With the development of industry and change of diet,fructose is widely used in various food products due to its sweetness and low cost,and its adverse effects have gradually been noticed due to the increase of fructose consumption in the last 30 years,especially the maternal fructose intake during pregnancy and lactation increases the appetite of offspring,however,its mechanism is still unclear.The hypothalamus senses the nutritional status of the body and integrates hormonal signals from the periphery,which is a key region in appetite regulation.Therefore,a fructose exposure model was established in pregnant and lactating female rats,and full-length RNA sequencing was performed using an Oxford nanopore technology platform to mine the DEGs affecting appetite in the hypothalamus of littermates according to the gene expression system,and to verify the protein expression levels of PI3 K,Akt,p-Akt and Fox O1 in the PI3K/Akt signaling pathway.Methods: Eighteen pregnant rats were randomly divided into 3 groups: control group,13%fructose group and 40% fructose group,6 rats in each group.From the day of gestation,drinking water with different concentrations of fructose was provided until the end of lactation,and all were given normal chow.The pups were kept in separate cages on day 21 and day 60,and deionized water without fructose was consumed.The expression of NPY in the hypothalamic arcuate nucleus of the three groups of pups was observed by immunofluorescence on day 21 and day 60,and the expression of Xprl,Kcnq3,Micall2,Prdm16 and Arhgap26,Traf3,Arf6,Tnc in the hypothalamus of the pups on day 60 was detected by RT-q PCR.The m RNA expression levels of Xprl,Kcnq3,Micall2,Prdm16 and Arhgap26,Traf3,Arf6 and Tnc in the hypothalamus tissues of the three groups of pups on day 21 and day 60 of life were detected by Western blot.Results: Body weight of both mothers and pups in all groups showed an increasing trend over time,and insulin and leptin levels,body weight,energy intake,and 12-h fasting glucose levels increased significantly in the fructose group compared with the control group;transcriptomic data identified 662 DEGs in F13 and 648 DEGs in F40,which are involved in food intake,hormone activity,signal transduction,and metabolism.Immunofluorescence results showed that NPY protein expression levels were significantly increased in the fructose group compared to Con,with the most significant increase in F40.RT-qPCR results showed that compared to Con,m RNA levels of Xprl,Kcnq3,Micall2,Arhgap26,Traf3 were significantly increased,and Prdm16,Arf6,Tnc m RNA levels were significantly decreased.In the Western blot results,the protein expression levels of PI3 K and p-Akt were significantly decreased and those of Fox O1 were significantly increased in the hypothalamus of the fructose group littermates compared with Con.Conclusion: The study of differentially expressed genes and changes in the expression of PI3K/Akt/Fox O1 signaling pathway in the hypothalamus of the litter confirmed that high fructose intake during gestation and lactation in the mother affected the regulation of appetite in the hypothalamus of the offspring,which may be associated with increased appetite in the litter.