## 4 Discussion
It has been indicated that flavonoids in mulberry leaves attenuate the development of atherosclerotic lesions by inhibiting the oxidative modification of LDL [53]. Persistent hyperglycemia triggers progressive damage to pancreatic \(\beta\)-cell function with enhanced IR, leading to significant metabolic disturbances in insulin target organs. The metabolic disturbances result in abnormally high blood glucose levels, creating a vicious cycle [54]. In the study, insulin secretion was increased in T2 DM mice, but the GSP remained at a high level, suggesting that T2 DM mice had severe IR. After FMLSP intervention, insulin and GSP levels were decreased, and the islet volume and morphology were also improved. Based on previous reports, it was hypothesized that the active ingredients (flavonoids, 1-DNJ, polysaccharides, etc.) in FMLSP could ameliorate IR by protecting the structure of pancreatic islet cells and enhancing insulin sensitivity [55]. [PERSON] et al. found that flavonoids in mulberry leaves could ameliorate IR by improving mitochondrial function in skeletal muscle [19]. Diabetic hyperglycemia boosts free radical generation in the body, resulting in oxidative stress. Conversely, it has been shown that oxidative stress plays a major role in the development of insulin resistance [56; 57]. It was reported that antioxidant enzymes (SOD, GSH-Px, and CAT) defend against the formation of reactive oxygen species in response to oxidative stress, while MDA production is a marker of oxidative stress leading to liver injury [33]. [PERSON] et al. discovered that the multi-components in mulberry leaves could relieve oxidative stress. This is because they can enhance the activity of antioxidant enzymes and reduce MDA accumulation. Consequently, IR is improved, and hypoglycemic effects are achieved [58]. The results of the present study are in agreement with the findings of [PERSON] et al. [58]. It was hypothesized that it was because the frosted mulberry leaves were superfinely crushed to expose a large number of active substances such as phenols, free polyphenols, and flavonoids bound to dietary fibers, which allowed them to be fully utilized, thus decreasing reactive oxygen species levels and exerting an antioxidant effect in T2 DM mice [59; 60]. The liver serves as a major organ involved in glycolipid metabolism, and disorders of glycolipid metabolism due to IR can impair liver function [47]. Notably, circulating ALT and AST maintain basal concentrations in physiological conditions yet exhibit marked elevation upon hepatocyte membrane integrity compromise, establishing these enzymes as sensitive biomarkers for quantifying hepatocellular injury severity [61]. Elevated ALT and AST levels combined with the morphologic structure of liver tissues in T2 DM mice suggested severe liver injury in T2 DM mice, similar to previous studies [62]. In the present study, FMLSP had a protective effect on liver tissue. Glycogen is the storage form of glucose, and a decrease in glycogen stores is associated with disturbances in glucolipid metabolism due to IR [63]. And glycolysis is a key stage that organisms must undergo to metabolize glucose, and hexokinase and pyruvate kinase are key enzymes in the glycolytic pathway, and they play an important role in maintaining blood glucose homeostasis [47]. In the current study, T2 DM impairs hepatic glycogen synthesis and attenuates glycolysis in mice. This means that glucose cannot be properly catabolized and metabolized, and therefore blood glucose levels are elevated [58]. FMLSP intervention improves glycolysis in T2 DM mice, reduces hepatic glucose output, promotes hepatic glycogen synthesis, and thus reduces blood glucose levels, which ultimately facilitates the restoration of glucose homeostasis in T2 DM mice. Emerging evidence delineates a pathological cascade wherein hyperglycemia induces gut microbiota dysbiosis, subsequently driving metabolic dysregulation and chronic low-grade inflammation that collectively exacerbate diabetes mellitus progression [64].