Metabolomics-guided exploration of Pinus wallichiana A.B. Jacks. cones for antidiabetic compounds: Integrating in vitro, in silico, in vivo, and histopathological approaches

Pinus wallichiana A.B. Jacks., also known as Himalayan white pine, is recognized in traditional medicine for its various uses, including diabetes mellitus (DM). Pinus species like Pinus roxburghii (Chir pine) and Pinus gerardiana (Chilgoza pine) have shown promise for antidiabetic properties. This study focuses on the use of edible cones of Pinus wallichiana (P. wallichiana) for the potential management of DM. The methanolic extract of Pinus wallichiana (Pw.Cme) was subjected to Gas Chromatography–Mass Spectrometry (GC-MS) analysis, total phenolic content (TPC) and total flavonoid content (TFC) analyses, and qualitative phytochemical studies. The Pw.Cme and its derived fractions were evaluated for their in vitro antioxidant, α-glucosidase, and α-amylase inhibitory studies, and the identified compounds were docked against enzyme targets, followed by Molecular Dynamics Simulation (MDS) studies. Detailed in vivo antidiabetic and histopathological studies were performed following standard procedures. The GC-MS analysis of Pw.Cme lead to identification of 45 compounds, and the Pinus wallichiana ethyl acetate (Pw.EtAc) fraction exhibited the highest TPC (258.55 mg gallic acid equivalent [GAE]/g) and TFC (63.05 mg quercetin equivalent [QE]/g). In 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assays, the respective highest anti-radical activity was observed for Pw.EtAc, as IC50=13.0 µg/mL and 5.3 µg/mL. In enzymes inhibition studies, considerable α-amylase inhibition was observed for Pw.EtAc with IC50=1.98 µg/mL, and IC50=7.2 µg/mL for α-glucosidase. In vivo studies indicated that the administration of Pw.EtAc resulted in a marked decrease in fasting blood glucose levels, hyperlipidemia, and weight loss in diabetic albino mice. Histopathology of vital organs of albino mice, administered with various doses of Pw.EtAc showed a healing effect against alloxan-induced lesions in the heart, pancreas, liver, and kidneys. In conclusion, we can claim that the extract of P. wallichiana cones is rich in many phytocomponents and have potential antidiabetic properties.