Figure 7

The effect of glabridin on the expression, deposition and arrangement of extracellular matrix in hypertrophic scar.

(A) Immunoblotting analysis on the expression of p-FAK-Tyr⁴⁰⁷, FAK, p-Src-Tyr⁵²⁹, p-Src-Tyr⁴¹⁶ and Src after treating hypertrophic scar fibroblasts with 100 μM glabridin. Each data point of the phosphorylated form was normalized against its corresponding total protein level with the value in DMSO (vehicle control) group arbitrarily set at 1. Each bar represents a mean ± SD of n = 4. ^*P < 0.05, ^**P < 0.01. (B) Co-immunoprecipitation was performed to examine the effect of glabridin on FAK-Src association in hypertrophic scar fibroblasts. Mouse origin anti-Src IgG was used as the precipitating antibody to extract interacting proteins binding to Src. Normal mouse (ms) IgG was used to replace anti-Src IgG and served as negative control, while HF lysates without co-immunoprecipitation served as positive control. Each data point of FAK was normalized against its corresponding Src level with the value in DMSO group arbitrarily set at 1. Each bar represents a mean ± SD of n = 4. ^**P < 0.01. (C) Immunoblotting analysis on the expression of Col I, Col III and α-SMA after treating hypertrophic scar fibroblasts with 100 μM glabridin. Each data point was normalized against its corresponding actin level with the value in DMSO group arbitrarily set at 1. Each bar represents a mean ± SD of n = 4. ^*P < 0.05. (D) H&E staining showed ECM deposition in hypertrophic scar tissue after glabridin treatment. Scale bar = 100 μm. The relative density of ECM deposition was analyzed by Image J software with the value in DMSO group arbitrarily set as 1. Each bar represents a mean ± SD of n = 4. ^**P < 0.01. (G) Masson staining showed collagen deposition as well as its arrangement. Scale bar = 100 μm. The relative density of collagen deposition was analyzed by Image J software with the value in DMSO group arbitrarily set as 1. Each bar represents a mean ± SD of n = 4. ^**P < 0.01.