Anti-Inflammatory Effect of Phytoncide in an Animal Model of Gastrointestinal Inflammation

Abstract

Background: Phytoncide is known to have antimicrobial and anti-inflammatory properties. Purpose: This study was carried out to confirm the anti-inflammatory activity of two types of phytoncide extracts from pinecone waste. Methods: We made two types of animal models to evaluate the efficacy, an indomethacin-induced gastroenteritis rat model and a dextran sulfate sodium-induced colitis mouse model. Result: In the gastroenteritis experiment, the expression of induced-nitric oxide synthase (iNOS), a marker for inflammation, decreased in the phytoncide-supplemented groups, and gastric ulcer development was significantly inhibited (p < 0.05). In the colitis experiment, the shortening of the colon length and the iNOS expression were significantly suppressed in the phytoncide-supplemented group (p < 0.05). Conclusions: Through this study, we confirmed that phytoncide can directly inhibit inflammation in digestive organs. Although further research is needed, we conclude that phytoncide has potential anti-inflammatory properties in the digestive tract and can be developed as a functional agent.

Keywords: Pinus koraiensis; anti-inflammation; colitis; gastritis; phytoncide.

Figure 1

Effect of phytoncide on an indomethacin-induced gastric ulcer. (A) Photograph of the stomach of a rat with indomethacin-induced gastritis, (B) gastric ulcer scoring graph of a rat with indomethacin-induced gastritis. G1, normal control; G2, Vehicle control (gastric ulcer control, indomethacin 80 mg/kg); G3, positive control (lansoprazole 30 mg/kg); G4, phytoncide oil 10 mL/kg; G5, phytoncide oil 5 mL/kg; G6, pine cone water extract 10 mL/kg; G7, pine cone water extract 5 mL/kg. Representative data are expressed as mean ± SEM of experiments (n = 10) for each group. *** p < 0.001 compared to vehicle control.

Figure 2

Effect of phytoncide treatment on histological changes in indomethacin-induced gastric damage. (A) Results of the H&E staining, × 100 magnification. (B) Effect of phytoncide on histological scores of the indomethacin-induced gastric ulcer. G1, normal control; G2, vehicle control (gastric ulcer control, indomethacin 80 mg/kg); G3, positive control (lansoprazole 30 mg/kg); G4, phytoncide oil 10 mL/kg; G5, phytoncide oil 5 mL/kg; G6, pine cone water extract 10 mL/kg; G7, pine cone water extract 5 mL/kg. Representative data are expressed as mean ± SEM of experiments (n = 10) for each group. ** p < 0.01 and *** p < 0.001 compared to vehicle control.

Figure 3

Immunohistochemical analysis: effect of phytoncide treatment on iNOS expression in indomethacin-induced gastric damage. (A) Photograph of stomach tissue from rat with indomethacin-induced gastritis, × 100 magnification. (B) iNOS expression graph of rat with indomethacin-induced gastritis. G1, normal control; G2, vehicle control (gastric ulcer control, indomethacin 80 mg/kg); G3, positive control (lansoprazole 30 mg/kg); G4, phytoncide oil 10 mL/kg; G5, phytoncide oil 5 mL/kg; G6, pine cone water extract 10 mL/kg; G7, pine cone water extract 5 mL/kg. *** p < 0.001 compared to vehicle control.

Figure 4

Effect of phytoncide treatment on symptoms of mice with DSS-induced colitis. (A) Effect of phytoncide treatment on body weight change in mice with DSS-induced colitis, (B) effect of phytoncide treatment in photograph of colon tissues of mice with DSS-induced colitis, (C) graphs of colon length, (D) effect of phytoncide treatment on disease activity index in mice with DSS-induced colitis. G1, normal control; G2, vehicle control (colitis control, DSS 2% in DW); G3, prevention phytoncide oil 10 mL/kg; G4, therapy phytoncide oil 10 mL/kg; G5, prevention pine cone water extract 10 mL/kg; G6, therapy pine cone water extract 10 mL/kg. Representative data are expressed as mean ± SEM of experiments (n = 10) for each group. ^#p < 0.05 compare to normal control. * p < 0.05 and ** p < 0.01 compared to vehicle control.

Figure 5

Effect of phytoncide treatment on histological changes on DSS-induced colitis damage. (A) Results of the H&E staining, × 100 magnification. (B) Effect of phytoncide on histological scores of DSS-induced colitis. G1, normal control; G2, vehicle control (colitis control, DSS 2% in DW); G3, prevention phytoncide oil 10 mL/kg; G4, therapy phytoncide oil 10 mL/kg; G5, prevention pine cone water extract 10 mL/kg; G6, therapy pine cone water extract 10 mL/kg. Representative data are expressed as mean ± SEM of experiments (n = 10) for each group. ** p < 0.01 compared to vehicle control.

Figure 5

Effect of phytoncide treatment on histological changes on DSS-induced colitis damage. (A) Results of the H&E staining, × 100 magnification. (B) Effect of phytoncide on histological scores of DSS-induced colitis. G1, normal control; G2, vehicle control (colitis control, DSS 2% in DW); G3, prevention phytoncide oil 10 mL/kg; G4, therapy phytoncide oil 10 mL/kg; G5, prevention pine cone water extract 10 mL/kg; G6, therapy pine cone water extract 10 mL/kg. Representative data are expressed as mean ± SEM of experiments (n = 10) for each group. ** p < 0.01 compared to vehicle control.

Figure 6

Result of immunohistochemical analysis: effect of phytoncide treatment on iNOS expression in DSS-induced colitis damage. (A) Photograph of colon from mouse with DSS-induced colitis, × 100 magnification. (B) iNOS expression graph of mouse with DSS-induced colitis. G1, normal control; G2, vehicle control (colitis control, DSS 2% in DW); G3, prevention phytoncide oil 10 mL/kg; G4, therapy phytoncide oil 10 mL/kg; G5, prevention pine cone water extract 10 mL/kg; G6, therapy pine cone water extract 10 mL/kg. ** p < 0.01 compared to vehicle control.