Fig 1.
Effect of calophyllolide on HaCaT and RAW264.7 cell viability.
HPLC chromatograms of the isolated calophyollide (A) and standard control (B). This compound was recorded at 233 nm, and its retention time is 36.6 min. (C) No effect of CP on the viability of both HaCaT and murine macrophage RAW264.7 cells after 24 h treatment.
Fig 2.
The enhancement of wound closure by calophyllolide.
Mice were daily treated with CP (6 mg/animal) and PI (100 mg/animal) until enthanasia. (A) Process of surgical wound healing in CP-treated group versus vehicle and PI-treated group, scale bar = 1cm (five animals per group). (B) The wound area (%) in the three treatment groups from day 1 to day 14 (n = 3–4 animals per group per experiment). (C) HE staining of cutaneous wound healing at day 7 and day 14 post-operation. Arrows indicate the wound sites, scale bar = 500 μm.
Table 1.
Effect of calophyllolide (CP) on acceleration of wound closure.
Fig 3.
Histological and quantitative analyses of the cutaneous wound healing of calophyllolide.
Mice were daily treated with CP (6 mg/animal) and PI (100 mg/animal) until enthanasia. (A) Histological observation of collagen on wound healing at day 14 by Masson’s Trichrome staining. Reduction of collagenous scar (arrow head) in CP-treated group compared to vehicle- and PI-treated groups. Arrows indicate wound site with scale bar = 1cm. (B) Representative graph of semi-quantitative collagen content at day 10 and day 14 (n = 3–4 animals per group per experiment). Data are represented as mean ± SEM and compared by one-way ANOVA.
Fig 4.
Characterization of spleen changes in wound healing.
Mice were daily treated with CP (6 mg/animal) and PI (100 mg/animal) until enthanasia. (A) Comparison of spleen size in each group at day 7 and day 14, scale bar = 1 cm. (B) Spleen length, (C) Spleen weight, (D) spleen index of each group at day 7 post-operation. (E) Comparison of total splenic cell numbers, live and dead of splenic cells in each group at day 7. Data are represented as mean ± SEM and compared by one-way ANOVA (n = 3–4 animals per group per experiment). * P<0.05, ** P<0.01, *** P<0.001.
Fig 5.
Effect of calophyllolide on myeloperoxidase (MPO) activity.
All mice were sacrificed on day 1 and day 5 post-operation, and skin tissue samples were collected to assess MPO activity (n = 3 mice per group per experiment). Data are represented as mean ± SEM and compared by one-way ANOVA. *** P<0.001.
Fig 6.
Attenuation of inflammatory cytokines expression by calophyllolide.
Serum levels of (A) IL-1β, (B) IL-6, (C) TNF-α, and (D) IL-10. Data are represented as mean ± SEM and compared by one-way ANOVA (n = 3 mice per group per experiment). * P<0.05, ** P<0.01, *** P<0.001.
Fig 7.
Expression of M1/M2 macrophage-related genes/markers during SLS induced inflammation.
(A and B): M1-related genes/markers (CD14 and CD127), (C and D): M2-related genes/markers (CD163 and CD206). Expression level was measured by qRT-PCR analysis.