Extract of Pinus densiflora needles suppresses acute inflammation by regulating inflammatory mediators in RAW264.7 macrophages and mice

Pharm Biol. 2022 Dec;60(1):1148-1159. doi: 10.1080/13880209.2022.2079679.

Abstract

Context: Pinus densiflora Siebold & Zucc. (Pinaceae) needle extracts ameliorate oxidative stress, but research into their anti-inflammatory effects is limited.

Objective: To investigate antioxidant and anti-inflammatory effects of a Pinus densiflora needles (PINE) ethanol extract in vitro and in vivo.

Materials and methods: We measured levels of reactive oxygen species (ROS), superoxide dismutase (SOD) and inflammatory mediators in lipopolysaccharide (LPS)-stimulated RAW264.7 cells at various PINE concentrations (25, 50 and 100 μg/mL; but 6.25, 12.5 and 25 μg/mL for interleukin-1β and prostaglandin E2 (PGE2)). Thirty ICR mice were randomized to six groups: vehicle, control, PINE pre-treatment (0.1, 0.3 and 1 mg/left ear for 10 min followed by arachidonic acid treatment for 30 min) and dexamethasone. The posttreatment ear thickness and myeloperoxidase (MPO) activity were measured.

Results: PINE 100 μg/mL significantly decreased ROS (IC50, 70.93 μg/mL, p < 0.01), SOD (IC50, 30.99 μg/mL, p < 0.05), malondialdehyde (p < 0.01), nitric oxide (NO) (IC50, 27.44 μg/mL, p < 0.01) and tumour necrosis factor-alpha (p < 0.05) levels. Interleukin-1β (p < 0.05) and PGE2 (p < 0.01) release decreased significantly with 25 μg/mL PINE. PINE 1 mg/ear inhibited LPS-stimulated expression of cyclooxygenase-2 and inducible NO synthase in RAW264.7 macrophages and significantly inhibited ear oedema (36.73-15.04% compared to the control, p < 0.01) and MPO activity (167.94-105.59%, p < 0.05).

Discussion and conclusions: PINE exerts antioxidant and anti-inflammatory effects by inhibiting the production of inflammatory mediators. Identified flavonoids such as taxifolin and quercetin glucoside can be attributed to effect of PINE.

Keywords: Reactive oxygen species; anti-inflammatory; antioxidant; arachidonic acid; ear oedema; lipopolysaccharide.

Publication types

  • Randomized Controlled Trial, Veterinary

MeSH terms

  • Animals
  • Anti-Inflammatory Agents / therapeutic use
  • Antioxidants / pharmacology
  • Dinoprostone / metabolism
  • Inflammation / pathology
  • Inflammation Mediators* / metabolism
  • Interleukin-1beta / metabolism
  • Lipopolysaccharides / pharmacology
  • Macrophages
  • Mice
  • Mice, Inbred ICR
  • Pinus*
  • Plant Extracts / therapeutic use
  • RAW 264.7 Cells
  • Reactive Oxygen Species / metabolism
  • Superoxide Dismutase / metabolism

Substances

  • Anti-Inflammatory Agents
  • Antioxidants
  • Inflammation Mediators
  • Interleukin-1beta
  • Lipopolysaccharides
  • Plant Extracts
  • Reactive Oxygen Species
  • Superoxide Dismutase
  • Dinoprostone

Grants and funding

This work was supported by the National Institute of Biological Resources (NIBR) funded by the Ministry of Environmental of the Republic of Korea under Grant No. NIBR201726101 and the Chung-Ang University Graduate Research Scholarship in 2021.