Anti‐Inflammatory Activity of Novel Dehydrozingerone‐6 (DHZ‐6) Through Modulation of iNOS, COX‐2, and NF‐κB/p65 in LPS‐Stimulated Macrophages and In Vivo Acute Lung Injury Murine Model

Acute lung injury (ALI) poses a significant threat in respiratory diseases and can lead to organ failure in chronic conditions. In this study, we evaluated the preventive effects of Dehydrozingerone‐6 against Lipopolysaccharide (LPS)‐induced inflammation in macrophages using RAW 264.7 and in a mouse model of ALI. During preliminary screening, Dehydrozingerone‐6 demonstrated significant inhibition of cytokines and maintained 97% cell viability at 10 μM. Further in vitro studies showed that Dehydrozingerone‐6 effectively suppressed LPS‐induced production of nitric oxide, interleukin‐6 (IL‐6), tumor necrosis factor‐alpha (TNF‐α), interferon‐gamma (IFN‐γ), interleukin‐1 beta (IL‐1β), and reactive oxygen species (ROS) at a concentration of 10 μM. Immunocytochemistry and Western blot analyses revealed reduced expression levels of inducible nitric oxide synthase (iNOS), cyclooxygenase‐2 (COX‐2), Nuclear Factor kappa B (NF‐κB/p65), and phosphorylated IκBα (p‐IκBα). In vivo LPS‐induced lung injury model revealed that Dehydrozingerone‐6 protected lung tissue from degradation and significantly inhibited IL‐6 and TNF‐α production at a dose of 50 mg/kg. Furthermore, Dehydrozingerone‐6 significantly reduced carrageenan‐induced paw edema and leukocyte migration. Dehydrozingerone‐6 also markedly reduced vascular permeability in an acetic acid‐induced model. Toxicological studies conducted at doses up to 2000 mg/kg body weight showed no notable alterations in hematological parameters or histopathology of the liver and kidney, suggesting a favorable safety profile. Overall, Dehydrozingerone‐6 exhibited strong anti‐inflammatory activity in vitro as well as in vivo studies by downregulating the generation of pro‐inflammatory mediators and oxidative stress through attenuation of iNOS, COX‐2, and NF‐κB/p65.