| Diabetes mellitus(DM)is a chronic disease caused by relative or absolute insufficiency of insulin,which leads to metabolic disorders such as blood glucose,protein and fat.Comprehensive and effective control of diabetes,as well as the use of oral drugs and insulin,requires further study to develop additional treatment options.Mulberry leaves are one of the most commonly used medicinal and herbaceous traditional Chinese medicines that are currently considered for the treatment of diabetes mellitus and its complications.The relevant researches indicated that the alkaloids,flavonoids,and polysaccharides in mulberry leaf impart regulatory effects on blood glucose levels.To identify the hypoglycemia-related active components in mulberry leaf and their mechanisms,the mice were used as experimental materials in this study.Mainly using the hyperglycemic mice model construction and the combination of transcriptomic and network pharmacology approaches,we investigated the physiological effects of mulberry leaf active components on hyperglycemia model mice according to physiological differences of the mice.The regulation effect of mulberry leaf active components on differentially expressed genes in hyperglycemic mice were revealed from the gene expression level,and the significantly expressed genes were verified by qRT-PCR and Western blotting.Meanwhile,GC/MS and pharmacological analysis were used to identify the mulberry leaf active components and analyze their anti-sugar targets.The main results of this experiment are as follows:(1)The statistical results of physiological differences in mice: After 10 weeks,the body weight of the mulberry leaf-treated mice was significantly higher than that of the hyperglycemic mice;the body weight of the mice that received the mulberry leaf powder was lower than that of normal mice.The blood glucose values were significantly decreased,and the serum insulin levels in the mulberry leaf treated group mice were significantly higher than that in the hyperglycemic mice.In general,the mulberry leaf could significantly improve the insulin sensitivity of hyperglycemic mice.It shows that mulberry leaf has a certain effect on anti-hyperglycemia.(2)The transcriptomic method was used to analyze the differentially expressed genes(DEGs)in liver tissue of normal mice(C group),hyperglycemic mice(DC group)and mulberry leaf treated mice(DD group).Among them,the total number of differentially expressed genes of DC group compared with C group were 679,of which 393 were upregulated and 286 were downregulated;the total number of differentially expressed genes of DD group compared with C group were 740,of which 353 were upregulated and 387 were downregulated;the total number of differentially expressed genes of DD group compared with CC group were 716,of which 211 were upregulated and 405 were downregulated.The GO analysis results showed that differentially expressed genes were mainly involved in stress response,metabolism and receptor binding,and the KEGG analysis results showed that differentially expressed genes were mainly involved in the metabolic pathway,inflammatory mediator regulation and insulin signal transduction.(3)The qRT-PCR and Western blotting results showed that the expression of LY6 A,GRB10,TAP1,IGF2,UBD protein were consistent with the transcription level.The mulberry leaf active components may play a great effect in reducing insulin resistance and restoring glucose metabolism by regulating the expression of these differential proteins.(4)Mulberry leaf active components identification and anti-sugar targets analysis: GC/MS method was used to determine the active components of mulberry leaf.A total of 202 active metabolites,mainly flavonoids and polyphenols,such as pentahydroxyflavone and chlorogenic acid were determined.Further pharmacological analysis of the active components indicated that 22 potential anti-glucose effects compounds that can regulate IRS1,TNF,TAP1,IGF2,FASN and other proteins,which are related to the insulin and inflammatory signaling pathways,glucose metabolism and other related pathways.Conclusion: The active components of mulberry leaves have certain effects on anti-hyperglycemia.It is shown that after feeding mulberry leaves in hyperglycemic mice,the active components of mulberry leaf can significantly reduce the body’s blood glucose and increase the serum insulin level.The mulberry leaf active components may alleviate the disorder of glucose metabolism and lipid metabolism caused by hyperglycemia by acting on the related proteins in insulin signaling pathway such as IGF2,TAP1,so as to reduce blood glucose and improve islet function.At the same time,it also laid a theoretical foundation for revealing the related biological effects of "multi-component-multi-target-multi-pathway-multi-disease" of mulberry leaf. |
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