Preliminary Study On The Protective Effect And Mechanism Of Hesperidin On Diabetes Based On Molecular Docking And Network Pharmacology

Background: Diabetes mellitus is a metabolic disorder caused by genetic and environmental factors,which is mainly manifested in insulin insensitivity,insulin deficiency and impaired insulin-related functions.Due to the high prevalence of the associated disability and mortality,diabetes mellitus has become one of the serious health problems worldwide.At present,the commonly used hypoglycemic drugs have different degrees of side effects.With the development of modern Chinese medicine research,researchers have found that more and more single herbs have the effects of improving diabetes mellitus and its complications.Hesperidin is a flavonoid compound and is also considered a vitamin P.It has a wide range of pharmacological effects,such as anti-inflammatory,preventing bleeding from ruptured capillaries,anti-oxidation and reducing blood viscosity.Clinically,it is mainly used for adjuvant treatment of cardiovascular system diseases.However,there are currently few studies on its protective effect on diabetes mellitus.In order to explore the effect of hesperidin on diabetes mellitus,this study explore whether DPP-4 is a potential target for hesperidin to protect diabetes mellitus based on molecular docking and network pharmacology.We found that hesperidin has a strong binding effect with DPP-4,and hesperidin can inhibit the DPP-4 enzyme activity in vitro,confirming that DPP-4 is the target of hesperidin.Inhibition of DPP-4 can increase the level of endogenous GLP-1 and promote the release of insulin from pancreatic β cells.In addition,it can also improve the function of pancreatic β-cells.Therefore,we researched whether hesperidin had a protective effect on pancreatic islet β cells and verified it through a series of experiments.The results show that hesperidin has a protective effect on the islet β-cell damage induced by high fatty acid and glucose.The findings of this study provide a certain theoretical basis for hesperidin to protect diabetes mellitus.1.To explore the target of hesperidin to protect diabetes mellitus based on molecular docking and network pharmacology and its in vitro verificationObjective:To explore that DPP-4 is a potential target for hesperidin to protect diabetes mellitus based on molecular docking and network pharmacology,and to verify it with in vitro experiments.Methods:The molecular docking method was used to study the binding effect of hesperidin and DPP-4.Pubchem was used to download the 3D structural formula of hesperidin.Pharm Mapper was used to predict the targets of hesperidin.OMIM and Gene Cards database were used to search the targets related to diabetes mellitus.Venn diagram was used to predict the potential targets of hesperidin in the treatment of diabetes mellitus.String database was used to analyze protein-protein interactions.Cytoscape 3.6.0 software was used to construct the "disease-drug-target" network.In vitro enzyme activity experiments verify the inhibitory effect of hesperidin on DPP4 in vitro.Results:The results of molecular docking showed that hesperidin and DPP-4have a strong docking effect,with an affinity of-12.8 kcal/mol.The analysis revealed 278 potential targets for hesperidin,5589 targets for diabetes mellitus,and 185 potential targets for hesperidin to protect diabetes mellitus.PPI and "disease-drug-target" network confirmed that DPP-4 is a potential target for hesperidin to protect diabetes mellitus.In vitro enzyme activity assays verified that hesperidin can inhibit DPP-4enzyme activity.Conclusion:Hesperidin can bind to DPP-4 and inhibit its enzyme activity in vitro.This may be one of its mechanisms to protect diabetes mellitus.2.The protective effect of hesperidin on injury induced by glucolipotoxicity in islet beta cellsObjective:To investigate the protective effect and mechanism of hesperidin on the injury of mouse pancreatic beta cells induced by high fatty acid and glucose.Methods:MIN6 cells were treated with high fatty acid and glucose after pretreatment of hesperidin.CCK-8 and Hoechst 33258 fluorescence staining were used to determine the proliferation and apoptosis of MIN6 cells.WB was used to detect the expressions of apoptosis-related proteins Bcl-2 and Bax.RT-PCR was used to detect the expressions of inflammatory factors TNF-α and IL-1β.ELISA was used to test the insulin secretion of pancreatic islets.Results:High fatty acid and glucose decreased the ratio of Bcl-2/Bax,increased the expression of inflammatory factors TNF-α and IL-1β and inhibited the insulin secretion of mouse pancreatic islets.After pretreatment of hesperidin,the cell viability and Bcl-2/Bax ratio of MIN6 increased,the expressions of inflammatory factors TNF-α and IL-1β decreased with varying degrees,and the insulin secretion of mouse pancreatic islets increased.Conclusion:Hesperidin inhibits the apoptosis of mouse pancreatic islet β cell line MIN6 induced by high fatty acid and glucose,reduces the secretion of inflammatory factors and improve the insulin secretion of pancreatic islets by inhibiting apoptotic signaling pathway and improving pancreatic islet cell function.