Results showed that mulberry leaves could significantly reduce backfat thickness (P<0.05) and increase intramuscular fat (IMF) content (P<0.05). Lipidomics analysis showed that mulberry leaves improved lipid profile composition and increased the proportion of triglycerides (TG). Interestingly, IMF content was positively correlated with acyl C18:2 and negatively correlated with C18:1 of differential TGs. In addition, cecal microbiological analysis showed that mulberry leaves could increase the abundance of bacteria such as UCG−005, Muribaculaceae_norank, Prevotellaceae_NK3B31_group, Limosilactobacillus, and so on. At the same time improve the L-tyrosine-baton rouge ester, Oleic acid methyl ester, 21-deoxycortisol, N-acetyldihydrosphingosine, Mulberrin relative abundance. In this study, we found that mulberry leaves could extremely significantly increasing the mRNA relative expression of lipoprotein lipase (LPL), fatty acid-binding protein 4 (FABP4) and peroxisome proliferators-activated receptor γ (PPARγ) in muscle (P<0.01), significantly increasing mRNA relative expression of diacylglycerol acyltransferase 1 (DGAT1) (P<0.05) and significantly decreasing the mRNA relative expression of acetyl CoA carboxylase (ACC) (P<0.05), and aignificantly upregulated hormone-sensitive triglyceride lipase (HSL), peroxisome proliferators-activated receptor α (PPARα) mRNA relative expression (P<0.05) in backfat. In addition mulberry leaves could increased serum leptin and adiponectin (P<0.01). Ultimately lead to up-regulation of IMF and down-regulation of backfat.