A Study On The Therapeutic Role Of Genipin In Diabetic Retinopathy And Its Mechanism

Background: Diabetes mellitus(DM)is a metabolic disorder caused by environmental and genetic factors.Diabetic retinopathy(DR),as the major and first complication of DM,is a mainly blinding eye disease leading to the vision loss caused by the interaction of genetic factors and environmental factors.Traditional Chinese medicine(TCM)is helpful in treating and preventing chronic diseases like DM.With the developing of new technologies,such as mass spectrum,c DNA microarray,RNA-seq,Real-Time Quantitative PCR and network pharmacology(NP),made predicting disease treatment targets by informations and previous studies possible.Based on different databases,we predicted an active ingredient,genipin,can be used to treat DR.Genipin is an active TCM compound used to treat diabetic retinopathy(DR).In this study,a network pharmacology(NP)-based approach was employed to investigate the therapeutic mechanisms underlying genipin administration in DR.Methods: The potential targets for DR were identified according to 9different databases.TCM database screening and NP were used to predict the potential active targets and pathways of genipin in DR.Cell viability was tested in vitro to determine the effects of different doses of glucose and genipin on h RMECs.CCK-8,wound healing,Transwell,tube-forming,reactive oxygen species(ROS),and other assay kits were used to detect the effects of genipin on h RMECs during high levels of glucose.In vivo,a streptozotocin(STZ)-mouse intraocular genipin injection(IOI.)model was used to explore the effects of genipin on diabetes-induced retinal dysfunction.Western blotting was performed to identify the cytokines involved in proliferation,apoptosis,angiogenesis,ROS,and inflammation.The protein expression of the AGEs/ RAGE pathways was also examined.Results: Approximately 26 types of TCM,and nearly 300 active ingredients,including genipin,were identified.The NP approach successfully identified the AGEs-RAGE pathways,with the UCP2 target,as key method of genipin in treating DR.In the in vitro and in vivo models,we discovered that high glucose increased cell proliferation,apoptosis,angiogenesis,ROS,and inflammation.However,genipin application regulated cell proliferation and apoptosis,inhibited angiogenesis,and reduced ROS and inflammation in the HRMECs exposed to high glucose.Furthermore,the retinal thickness in the genipin-treated group was lower than that in the untreated group.AGEs/ RAGE signaling was increased by high glucose levels;however,genipin treatment decreased AGEs/ RAGE pathway expressions.Genipin was found to protect HG-induced h RMECs and the retina of STZ-mice,based on;1.the inhibition of UCP2 and GLUT1 decreased intracellular glucose,and glycosylation;2.Inhibit UCP2 leaded oxidative phosphorylation from ATP synthesis decreased oxidoreductase activity,and;3.The effect of glycosylation on vascular endothelial growth factor(VEGF),MMP9,and SCG3.Conclusion: Based on NP,we demonstrated the potential targets and pathways of genipin in the treatment of DR and confirmed its effective molecular mechanism in vitro and in vivo.Genipin protects cells and tissues from high glucose levels by regulating phosphorylation and glycosylation.Our study provides new insights into the key genes and pathways associated with the prognosis and pathogenesis of DR.