Figure 4
From: Connective tissue growth factor is correlated with peritoneal lymphangiogenesis
Connective tissue growth factor (CTGF) expression was correlated with expression of vascular endothelial growth factor-C (VEGF-C) and lymphatics in a rat diaphragmatic fibrosis model induced by chlorhexidine gluconate (CG). Diaphragmatic fibrosis was induced by intraperitoneal injection of CG in rats. Control rats were treated with saline. (a,b) Immunohistochemistry (IHC) showed that expression of CTGF, VEGF-C, and lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1) was increased in the rat diaphragmatic fibrosis model induced by CG compared with controls. The staining intensity for CTGF and VEGF-C was scored as follows: 0, absent; 1, mild; 2, moderate; 3, extensive. CTGF messenger RNA (mRNA) was detected in the CG model with in situ hybridization (ISH). Arrowheads indicate CTGF mRNA expression. Arrows indicate LYVE-1-positive lymphatic vessels. Insets show magnification of the dotted-line boxed areas. Scale bars; 100 μm. (c) CTGF, VEGF-C, LYVE-1, and podoplanin mRNA expression were increased in the diaphragm in CG-injected rats compared with controls as seen with quantitative polymerase chain reaction analysis. (d–f) IHC analysis showed positive correlations among CTGF, VEGF-C, and LYVE-1 expression in the CG model. Spearman’s rank correlation was used for the analysis. (g) Double immunofluorescent staining showed that CTGF expression was increased around LYVE-1-positive lymphatic vessels in the CG model. Bar graphs show means ± SD (Control, n = 5; CG model, n = 9).
