LC-MS/MS analysis and network pharmacology of Trigonella foenum-graecum - A plant from Ayurveda against hyperlipidemia and hyperglycemia with combination synergy

Background: The seed of Trigonella foenum-graecum L. (Methika in Sanskrit) is a well known kaphahara (balancing kapha) herb in Ayurveda indicated in Prameha or early diabetes mellitus. It is also useful in obesity and reduces lipid level of blood. Purpose: We aimed to explore the metabolites present in the plant extract and to establish the combination synergy and the network pharmacology along with the underlying the mechanism of action involved. Study design: LC-MS/MS based metabolite screening followed by ADME screening and finally network pharmacology exploration of the mechanism of action involved against hyperlipidemia and hypolipidemia with neighbourhood based combination synergy approach. Methods: Ethanolic extract of Trigonella foenum-graecum L. (TFHE) was subjected to LC-MS/MS analysis to identify the active constituents. Oral bioavailability and drug likeness was screened for all the compounds. Databases- Binding DB, DAVID, KEGG and STRING were used to gather information to develop the networks. The networks were constructed using Cytoscape 3.2.1. Combination synergy analysis was performed with the help of Cytoscape network analyzer tool with neighbourhood approach. Results: The LC-MS/MS analysis identified 13 compounds which were found to be bio-available and drug like following the QED and Veber drug likeness parameters. The pathway analysis showed enrichment for different pathways like MAPK pathway (p-4.69E-07), JAK-STAT pathway (p-6.30E-05), Adipocytokine (p-0.00179), Type 2 Diabetes mellitus (0.00441), Insulin signalling pathway (p-0.0121), mTOR signalling pathway (p-0.000378), which are all connected to hyperlipidemia and hyperglycemia. The combination synergy network identified 23 targets interacting with 13 compounds based on a network neighbourhood approach. Conclusion: The network pharmacology analysis strongly suggested the multimode evidences that TFHE largely works on the insulin signalling pathway and mainly based on its antioxidant potential due to its interaction with carbonic anhydrase. Various compounds were found to be interacting with key proteins that activates EGFR/AKT/mTOR signalling cascade which has therapeutic implication in hyperglycemia and hyperlipidemia. The combination synergy network analysis based on neighbourhood approach can help us in further understanding mechanism of multi-molecular fixed dose combinations.