Research on saponin active compounds of Tuchao Baibiandouren for the treatment of type-2 diabetes based on UHPLC-Q-Exactive Orbitrap MS and network pharmacology

Objective To explore the pharmacological mechanism of active saponin compounds of Tuchao Baibiandouren(Lablab Semen Album fried with earth,TCBBDR)in treating type 2 diabetes(T2DM)using UHPLC-Q-Exactive Orbitrap MS and network pharmacology.Methods UHPLC-Q-Exactive Orbitrap MS was used for a qualitative analysis of saponin compounds in TCBBDR.PharmMapper and CTD were used to screen drug active compounds and disease targets,and an active compound-target network was constructed via Cytoscape 3.8.0.Molecular docking was applied with the drug active compounds and key targets using AutoDock Vina 1.1.2,and a trajectory for the molecular dynamics simulation was completed by GROMACS 2019-3.Results Sixteen saponin compounds were identified from TCBBDR,along with 292 saponin compoud targets and 792 T2DM targets.Through Venn analysis of target saponin constituents and T2DM related targets,a total of 91 intersection targets were screened out in the treatment of T2DM with saponin.The mean values of degree,betweenness centrality and closeness centrality were taken as the thresholds to screen out 22 key genes,among which 4 key proteins namely MAPK1,IGF1 EGFR,PIK3R1 were selected in the top 10 key genes.On this basis,the saponin active constituent-target-signaling pathway network was established.The Gene Ontology(GO)enrichment analysis showed that the related biological modules included activity of steroid hormone receptor,steroid binding,and insulin receptor binding,etc.;the related signaling pathways were EGFR,PI3K-Akt and MAPK,etc.;regulating signaling pathways like MAPK could induce the proliferation,inhibition and apoptosis of pancreaticβcells,increase the quantity of pancreaticβcells,improve the functions of pancreaticβcells and stimulate the insulin secretion.Docking experiment analysis showed that all selected saponin compounds could enter the active sites of targets and form 3–14 hydrogen bonds with residues of the active sites.Moreover,van der Waals forces were present between chemical compounds and active sites.By combining the docking binding energy,we determined that the chemical compounds showed strong binding energy to the targets.Conclusion TCBBDR exerts therapeutic effects on diabetes through multi-compound and multi-target collaboration.Specifically,saponin components mediate pathways including inflammatory reaction and signal transduction to treat T2DM by regulating several key proteins that interact with EGFR and a series of signaling pathways related to disease development.

Screening and functional evaluation of the glucose-lowering active compounds of total saponins of Baibiandou(Lablab Semen Album)

Objective To screen forα-glucosidase inhibitor active compounds in the total saponins of Baibiandou(Lablab Semen Album)based on UHPLC-Q-Exactive Orbitrap MS technology and to evaluate its hypoglycemic activity in vivo.Methods Acarbose was used as the positive control,and the median inhibitory concentration(IC50)was used as the evaluation index ofα-glucosidase inhibitory activity to establish an in vitroα-glucosidase inhibition model.Further,UHPLC-Q-Exactive Orbitrap MS technology was used to screen and identify the active compounds ofα-glucosidase inhibitors in the total saponins of Baibiandou(Lablab Semen Album)in order to further verify the activity of the main active monomer and to perform homologous modeling and molecular docking of yeast-derivedα-glucosidase and human-derivedα-glucosidase,while the hypoglycemic activity was evaluated in diabetic mice.Results This study successfully identified 15 compounds with potentialα-glucosidase inhibitory activity,including Chikusetsusaponin IVa,from the total saponins of Baibiandou(Lablab Semen Album).Simultaneously,we verified the activity of the main active monomer Chikusetsusaponin IVa,and showed that it has strongα-glucosidase inhibitory activity.Theα-glucosidase inhibitory concentration IC50 was(565.2±1.026)μg/m L,and the IC50 of acarbose,which was lower than the positive control,was(706.6±1.058)μg/m L.The docking energies of Chikusetsusaponin IVa were–6.1 and–7.7 kcal/mol with yeast-derivedα-glucosidase and human-derivedα-glucosidase molecules,respectively.Both showed strong binding activity,and the levels of alanine aminotransaminase(ALT),aspartate aminotransaminase(AST),UREA,Creatinine(CREA),and cholesterol(CHO)were significantly decreased by Chikusetsusaponin IVa(P<0.05).In addition,it could repair damaged liver and pancreas cells of diabetic mice to some extent.Conclusion This study provides a basis for screeningα-glucosidase inhibitors and structural modifications of the total saponins of Baibiandou(Lablab Semen Album).