Unravelling the prophylactic anti-inflammatory potential of Koenigia tortuosa through modulation of cytokine levels and inflammatory markers in LPS-induced localized inflammation in Wistar rat models.

Abstract

Chronic inflammation, a pivotal factor in various chronic diseases, necessitates safe and effective treatments to alleviate disease severity and symptoms. Current interventional approaches, including synthetic steroids and non-steroidal anti-inflammatory drugs, pose safety concerns. Consequently, people seek plant-based alternatives as safer substitutes. Koenigia tortuosa, a medicinal plant with rich folklore claims, traditionally treats joint pain, swelling, dysentery and kidney related problems but lacks documentation. This study investigated anti-inflammatory properties of Koenigia tortuosa. Soxhlet extraction method was employed to obtain five different extracts of Koenigia tortuosa viz., hexane (95%), ethyl-acetate (99%), ethanol (99%), methanol (95%) and aqueous. Anti-inflammatory potential of different extracts was determined by both in vitro (including protein denaturation, nitric-oxide scavenging, proteinase inhibition, and erythrocyte membrane stabilization) and in vivo by performing histopathological studies and determining levels of various inflammatory markers like IL-1β, IL-6, IFN-γ and TNF-α using ELISA and, iNOS, PPAR-γ and COX-2 by Western blotting. GC-MS analysis was performed to reveal the bioactive compounds in extracts. At 600 μg/mL, two extracts, ethyl acetate and methanolic extract exhibited maximum inhibition of protein denaturation 75.07% ± 3.28% and 64.97% ± 1.73%, nitric oxide activity 88.06% ± 3.49% and 82.09% ± 3.61%, proteinase activity 82.06% ± 2.98% and 71.06% ± 3.58%, and erythrocyte-membrane haemolysis 84.94% ± 4.14% and 72.97% ± 4.68%, respectively (P < 0.001). In vivo studies using Wistar rats demonstrated no toxic effects of ethyl acetate and methanolic extract upon oral administration. These two extracts modulated cytokine levels and inflammatory markers, showing concentration dependent reductions in levels of IL-6, IL1-β, IFN-γ, TNF-α (P < 0.001), iNOS, COX-2 in LPS -induced inflammation in Wistar rats. At a dose of 100 mg/kgbwt, KTEA administration resulted in a substantial decrease in cytokine levels: IL1β from 68.99 ± 1.83 pg/mL to 31.68 ± 1.90 pg/mL (P < 0.001), IL6 from 80.40 ± 0.70 pg/mL to 39.47 ± 1.85 pg/mL (P < 0.01), TNFα from 71.34 ± 2.35 pg/mL to 29.37 ± 2.20 pg/mL (P < 0.001), and IFNγ from 120.27 ± 4.26 pg/mL to 68.07 ± 2.78 (P < 0.01) pg/mL. Similarly, a concentration dependent decrease in prostaglandins (273.68 pg/mL and 418.96 pg/mL by ethyl acetate and methanolic extract at 100 mg/kgbwt) and leukotrienes (239.37 pg/mL and 302.19 pg/mL by ethyl acetate and methanolic extract at 100 mg/kgBwt) were observed as compared with the LPS induced group (prostaglandins 1129.99 pg/mL and leukotrienes 558.67 pg/mL). We also observed that Koenigia tortuosa extracts improves the levels of lymphocytes and leukocytes. Notably, PPAR-γ expression exhibited a concentration dependent increase, suggesting potential anti-inflammatory effects through nuclear receptor modulation. Histopathological investigations demonstrated significant healing effects of extracts. Analysis using GC-MS unveiled the presence of bioactive compounds with potent anti-inflammatory properties. These findings suggest Koenigia tortuousa's anti-inflammatory mechanisms and potential therapeutic applications.