Figure 2.

Inhibition by chrysin of glucose-induced cellular induction of α-SMA and fibroblast specific protein (FSP)-1 (A) and secretion of collagen I and collagen IV (B) in mesangial cells and α-SMA induction in mouse glomeruli (C). Human renal mesangial cells were challenged for 3 days with 5.5 mM glucose, 5.5 mM glucose plus 27.5 mM mannitol as osmotic controls or with 33 mM glucose in the absence and presence of 1–20 μM chrysin. Cellular induction of α-SMA and FSP-1 and secretion of collagen I and collagen IV into culture media were measured by using Western blot analysis with a primary antibody against α-SMA, FSP-1, collagen I or collagen IV (A,B). The bar graphs (mean ± SEM, n = 3 independent experiments) in the bottom panel represent densitometric results obtained from Image analysis. β-actin protein was used as an internal control. Diabetic mice were orally administrated with 10 mg/kg chrysin for 8 weeks. The α-SMA induction in diabetic mouse glomeruli was visualized as FITC-green staining and nuclear counter-staining was done with the blue stain DAPI (C). Scale bar = 25 μm. The relative staining intensity was measured and the respective values are expressed as mean ± SEM (n = 4 mice in each group). Values in bar graphs not sharing a same lower case indicate significant different at p < 0.05.