Mechanisms of Action of Potentilla discolor Bunge in Type 2 Diabetes Mellitus Based on Network Pharmacology and Experimental Verification in Drosophila

Abstract

Purpose

Type 2 diabetes mellitus (T2DM) is associated with reduced insulin uptake and glucose metabolic capacity. Potentilla discolor Bunge (PDB) has been used to treat T2DM; however, the fundamental biological mechanisms remain unclear. This study aimed to understand the active ingredients, potential targets, and underlying mechanisms through which PDB treats T2DM.

Methods

Components and action targets were predicted using network pharmacology and molecular docking analyses. PDB extracts were prepared and validated through pharmacological intervention in a Cg>InR^K1409A diabetes Drosophila model. Network pharmacology and molecular docking analyses were used to identify the key components and core targets of PDB in the treatment of T2DM, which were subsequently verified in animal experiments.

Results

Network pharmacology analysis revealed five effective compounds made up of 107 T2DM-related therapeutic targets and seven protein–protein interaction network core molecules. Molecular docking results showed that quercetin has a strong preference for interleukin-1 beta (IL1B), IL6, RAC-alpha serine/threonine-protein kinase 1 (AKT1), and cellular tumor antigen p53; kaempferol exhibited superior binding to tumor necrosis factor and AKT1; β-sitosterol demonstrated pronounced binding to Caspase-3 (CASP3). High-performance liquid chromatography data quantified quercetin, kaempferol, and β-sitosterol at proportions of 0.030%, 0.025%, and 0.076%, respectively. The animal experiments revealed that PDB had no effect on the development, viability, or fertility of Drosophila and it ameliorated glycolipid metabolism disorders in the diabetes Cg>InR^K1409A fly. Furthermore, PDB improved the body size and weight of Drosophila, suggesting its potential to alleviate insulin resistance. Moreover, PDB improved Akt phosphorylation and suppressed CASP3 activity to improve insulin resistance in Drosophila with T2DM.

Conclusion

Our findings suggest that PDB ameliorates diabetes metabolism disorders in the fly model by enhancing Akt activity and suppressing CASP3 expression. This will facilitate the development of key drug targets and a potential therapeutic strategy for the clinical treatment of T2DM and related metabolic diseases.

Graphical Abstract

Keywords:

• Potentilla discolor Bunge
• T2DM
• network pharmacology
• molecular docking
• Drosophila

Abbreviations

T2DM: type 2 diabetes mellitus; PDB: Potentilla discolor Bunge; IL1B: Interleukin-1 beta; IL6: Interleukin-6; AKT1: RAC-alpha serine/threonine-protein kinase; TP53: Cellular tumor antigen p53; TNF: Tumor necrosis factor; CASP3: Caspase-3; DM: diabetes mellitus; IR: insulin resistance; TCM: traditional Chinese medicine; InR: insulin receptor; OB: oral bioavailability; DL: drug-likeness; PPI: protein-protein interaction; GO: gene ontology; KEGG: Kyoto encyclopedia of genes and genomes; HPLC: high-performance liquid chromatography; PBS: phosphate-buffered saline; TAG: triglycerides; Blue-9: blue FCF; CST: Cell signaling technology; WB: Western blotting; VEGFA: Vascular endothelial growth factor A; PDW: water extract of Potentilla discolor Bunge; HSD: high-sugar diet.

Ethical Statement

Network pharmacology analysis utilized publicly available data and was approved by the Ethics Committee of North China University of Science and Technology. The use of Drosophila melanogaster, which are invertebrates, is not considered of ethical concern. The animal study was audited by the Laboratory Animal Welfare Ethics Committee of North China University of Science and Technology in accordance with the local legislation and institutional requirements.

Acknowledgments

We would like to thank the Bloomington Drosophila Stock Center for providing stock of Drosophila melanogaster, members of the Wu lab and Li lab, Dr. Zhen Chen (Oriental Herbs KFT of Hungary) for discussion and critical comments, and Editage (http://www.editage.cn) for English language editing. This work was supported by the Hebei Natural Science Foundation (H2022209027), Tangshan Science and Technology Project (21130230C) and Tangshan Talent Funding Project (A202203021) to Chenxi Wu, the Science and Technology Partnership Program, Ministry of Science and Technology of China (KY201904005) and Hebei Natural Science Foundation (H2023209038) to Ji-an Li, the Hebei Natural Science Foundation (H2022209031) to Hong Chang, and Graduate Student Innovation Fund of North China University of Science and Technology (2023S15) to Yinghong Li.

Author Contributions

All authors made a significant contribution to the work reported, whether that is in the conception, study design, execution, acquisition of data, analysis and interpretation, or in all these areas; took part in drafting, revising or critically reviewing the article; gave final approval of the version to be published; have agreed on the journal to which the article has been submitted; and agree to be accountable for all aspects of the work.

Disclosure

The authors report no conflicts of interest in this work.