LC-MS profiling and multi-target mechanistic insights of Hibiscus rosa-sinensis in diabetes: Network pharmacology, molecular docking, MD simulation, PCA, and in-vitro α-amylase inhibition

Abstract

Background

Hibiscus rosa-sinensis Linn. (Zhū Jǐn Huā), known in Traditional Chinese Medicine (TCM) as Dà Hóng Huā, has been widely used for its cooling, detoxifying, and anti-inflammatory properties. It is a medicinal plant traditionally used for managing diabetes, yet its multi-target mechanisms remain largely unexplored. This study aimed to investigate the anti-diabetic potential of H. rosa-sinensis through LC-MS-based phytochemical profiling, network pharmacology, molecular docking, molecular dynamics (MD) simulations, principal component analysis (PCA), and in vitro α-amylase inhibition assay.

Methods

LC-MS analysis was performed to identify bioactive compounds present in the methanolic extract of H. rosa-sinensis. Identified compounds were subjected to network pharmacology analysis to predict potential diabetes-related protein targets. Key compound-target interactions were evaluated using molecular docking (AutoDock Vina v1.2.0), followed by 100 ns MD simulations to assess structural stability and binding dynamics. PCA and free energy landscape (FEL) analyses were conducted to examine conformational behaviour. Finally, in vitro α-amylase inhibition was tested to validate the hypoglycaemic potential.

Results

Quercetin demonstrated strong binding affinities with PTK2 (–8.2 kcal/mol), SRC (–9.2 kcal/mol), and α-amylase (–8.9 kcal/mol), suggesting its multi-target action in diabetes. MD simulation confirmed the structural stability of all three complexes over 100 ns, supported by consistent RMSD and favourable interaction profiles. The PCA and FEL analyses showed minimal conformational fluctuations and energetically favourable states. The in vitro α-amylase inhibition assay further validated the antidiabetic potential, with an IC₅₀ value of 107.75 µg/mL for the H. rosa-sinensis flower extract.

Conclusion

This integrative study provides strong evidence for the multi-target antidiabetic potential of Hibiscus rosa-sinensis, primarily attributed to quercetin and its derivatives. The findings highlight stable interactions with key diabetic targets and demonstrate significant α-amylase inhibition, supporting the traditional use of H. rosa-sinensis as a complementary therapy for diabetes management.