| Objective:A combination of network pharmacological exploration and in vivo experimental validation was used to investigate to verify the mechanism of action of Buyang Huanwu Decoction in the protection of type 2 diabetic mouse myocardium.Methods:1.Network pharmacology study: Searching for the active ingredients and corresponding BYHWD target genes using publicly available databases.Targets related to diabetic cardiomyopathy were collected and drug and disease targets were interleaved.These core targets were screened,enriched,and analyzed to construct a "drug-component-target pathwaydisease" network.Molecular docking enables validation of key active ingredients and central targets.2.In vivo experiments: Sixty SPF-grade male c57BL/6J mice,10 as a control group,the remaining 50 to establish the T2DM model,mice were fed a high fat diet for 4 weeks and then injected intraperitoneally with STZ,after successful modeling,split into 5 groups of 10 mice each.During the rearing period,general condition was monitored regularly,and each group was gavaged separately for a period of 8 weeks and material was collected afterwards.Serum INSR levels were measured by ELISA,and mouse cardiological changes were detected by HE and Masson staining.We measured the expression of insulin signaling pathway-related proteins in mouse cardiac tissue by Western Blot.Results:1.Network pharmacology study: 62 active ingredients of Buyang Huanwu Decoction were obtained,with a total of 469 active targets and 109 targets intersecting with diabetic cardiomyopathy.A total of 2077 biological processes,45 cellular components,158 molecular functions and 175 signaling pathways related to diabetic cardiomyopathy were enriched.The active ingredients baicalin and β-sitosterol showed high stability in binding to the target AKT1.2.Experiments in vivo: At 8 weeks post-treatment,mice in the model group were in a generally poorer state of health than those in the control,with significantly higher levels of blood glucose(P<0.01),lower body weight(P<0.05),higher food intake and water intake(P<0.01),and severe pathological damage to the heart tissue of the mice.And,in the present study,there was no significant change in blood glucose.Blood glucose levels were lower in mice from the BYHWD low,medium,and high dose and Metformin groups(P<0.01).Body weight,food intake,and water intake improved to different extents when compared to mice in the model group(P<0.05 or P<0.01).The level of serum INSR,the expression of PI3 K,AKT,and their phosphorylated proteins in cardiac tissues were decreased in the model group compared with the control group,which did not show a significant difference(P<0.01),and pGSK3β protein expression was significantly up regulated(P<0.01).There was a trend for the serum INSR level to increase in each therapeutic group compared to the model group(P<0.05),and both p-PI3 K and p-AKT expression were significantly increased(P<0.01),the levels of GSK3β and phosphorylated proteins were down regulated to different extent(P<0.01).Conclusion:1.Buyang Huanwu Decoction may exert its therapeutic effects on diabetic cardiomyopathy by regulating multiple signaling pathways,including insulin signaling pathway,and its core target molecules may include AKT and GSK3β.2.Buyang Huanwu Decoction can lower fasting blood glucose levels,and increase body mass,water intake,and consumption.Ameliorate myocardial pathomorphological changes,reduce myocardial fibrosis in T2DM mice,and have been shown to have a protective effect on the myocardium in the T2DM model.3.Buyang Huanwu Decoction may activate the insulin signaling pathway,upregulate INSR,p-PI3K and p-AKT,and downregulate p-GSK3β to improve the level of insulin transmission signals in cardiac tissues,thus exerting a protective effect on the myocardium of T2DM mice. |
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