The mechanism study of sleep disorders accelerating obesity by regulating the leptin/AMPK/SIRT1/Src/ACSS2/PPARγ pathway–mediated white fat synthesis

Sleep disorders, as a common health issue in modern society, are closely associated with an increased risk of obesity. This study aimed to investigate the impact of sleep disorders on white fat synthesis and their specific molecular mechanisms. First, the bulk RNA-seq dataset GSE299158 was utilized for differential expression gene screening and functional enrichment analysis to identify pathways associated with sleep disorders. Concurrently, the public single-cell transcriptome dataset GSE213496 was analyzed to characterize preadipocyte subpopulations, with differentiation potential assessed using CytoTRACE, oxidative stress scores calculated, pseudo-temporal trajectories reconstructed, and intercellular communication networks elucidated. Twenty-four ICR male mice were randomly and equally divided into four groups: control group, sleep disorder group, high-fat diet (HFD) group, and sleep disorder + HFD group. The mice were weighed, and white fat was excised to observe the white fat status in each group. The pituitary glands of the mice were removed, and protein expression was detected using Western blot. In vitro experiments were conducted using purchased 3T3-L1 mouse preadipocytes, which were subjected to different treatments, and protein expression was detected using Western blot. Bulk RNA-seq analysis revealed that leptin expression was significantly downregulated in mice with sleep disturbances, and differentially expressed genes were enriched in pathways related to oxidative stress, lipid metabolism, and immunity; Single-cell transcriptomics further revealed that sleep deprivation treatment induced significant transcriptional remodeling in preadipocytes, characterized by elevated oxidative stress scores, reduced differentiation potential (CytoTRACE), altered circadian trajectories, downregulation of mature adipocyte markers (Lpl, Fasn), and upregulation of fibrosis-related genes (Pdgfrb, Vim). Mice with sleep disorders showed significant weight gain and accumulated large amounts of white fat. Leptin and p- AMPK/t-AMPK expression were reduced, while NOX4, p-Src/t-Src, p-ACSS2/t-ACSS2, and nuclear PPARγ expression were increased. When sleep disorder mice were fed with HFD, their weight increased significantly, and they accumulated extremely large amounts of white fat. Leptin and p-AMPK/t-AMPK expression were significantly reduced, while NOX4, p-Src/t-Src, p-ACSS2/t-ACSS2, and nuclear PPARγ expression were increased. In vitro experiments showed that white fat inhibited the expression of p- AMPK/t-AMPK, SIRT1, PGC-1α, and UCP1, while promoting the expression of NOX4, p-Src/t-Src, p-ACSS2/t-ACSS2, nuclear PPARγ, C/EBPα, and SIK2. Leptin, on the other hand, promoted the expression of p- AMPK/t-AMPK, SIRT1, PGC-1α, and UCP1, while inhibiting the expression of NOX4, p-Src/t-Src, p-ACSS2/t-ACSS2, nuclear PPARγ, C/EBPα, and SIK2. Sleep disorders accelerate obesity progression by regulating white fat synthesis through the Leptin/AMPK/SIRT1/Src/ACSS2/PPARγ pathway.