Exploring the Ameliorative Role of Equisetum diffusum D. Don Whole-Plant Methanolic-extract in Acute Inflammation and Molecular Docking Analysis of GC-MS-identified Phytocompounds with Few Prominent Inflammatory Markers/Cytokines for Inspecting the Potent

Background: Equisetum diffusum D. Don (Fam. Equisetaceae), an important medicinal pteridophyte species has been traditionally used in the treatment of bone fracture, bone dislocation, and arthritis by various tribal communities of India. Objectives: To validate the potent anti-inflammatory properties of the whole plant methanolic-extract (EDME) using in silico, in vitro, and in vivo strategies. Materials and Methods: The EDME was characterized through total phenolic, flavonoid, tannin, saponin content and five antioxidant assays. The GC-MS analysis was done to screen the active phyto-constituents followed by in silico molecular docking studies with inflammatory markers. The in vitro anti-inflammatory activity was assessed by measuring protein denaturation-inhibition test, heat-induced and hypotonicity-induced haemolysis test. Carrageenan-induced paw-edema test was used to determine the in vivo anti-inflammatory activity of EDME. Results: Our results showed quite higher tannin content in EDME compared to saponin content, and showed satisfactory phenol and flavonoid contents. From the antioxidant survey, it was found that EDME has the highest activity against ABTS radical and the least against superoxide radical. Furthermore, GC-MS analysis of EDME reported the presence of 47 phyto-compounds, out of which few compounds showed good inhibition against Cyclooxygenase-2 (Cox-2) and IL-6 compared to standard NSAIDs, in molecular docking analysis. In vitro anti-inflammatory studies indicated that EDME inhibits protein denaturation, heat-induced, and hypotonicity-induced haemolysis significantly (p<0.001) compared to standard drugs. The in vivo anti-inflammatory study of the plant showed 52.26% and 73.36% reduction in paw-edema in both protective-dose groups respectively, when compared to the carrageenan control group. Conclusion: Our findings established the anti-inflammatory roles of the whole plant methanolic extract of Equisetum diffusum on strong ground which may lead to drug development for the treatment of inflammation-related complications.