Journal: Biomolecules

Article Title: Ursolic Acid Inhibits Collagen Production and Promotes Collagen Degradation in Skin Dermal Fibroblasts: Potential Antifibrotic Effects.

doi: 10.3390/biom15030365

Figure Lengend Snippet: Figure 3. UA induces MMP-1 by activation of MAPK pathways in primary dermal fibroblasts. Cells were treated with UA (10 µM) for the indicated time periods. (A) ERK1/2 activation, (B) c-Jun mRNA, (C) c-Jun protein, (D) c-Fos mRNA, and (E) c-Fos protein levels. (F) UA increases AP-1 reporter activity. Cells were transiently transfected with AP-1 reporter construct and AP-1 activity was determined by luciferase assay. (G,H) UA-induced MMP-1 expression was blocked by MAPK inhibitors. Cells were treated with MAPK (PD98059 and SP600125 for 2 h and then treated with UA (10 µM) for 24 h. (G) MMP-1 mRNA and (H) MMP-1 protein. mRNA levels were quantified by real-time RT-PCR and normalized to 36B4 mRNA, a ribosomal protein used as an internal control. Protein levels were determined by Western blot analysis and normalized to β-actin as a loading control. Data are expressed as mean ± SEM (N = 3). p-values are shown in the respective graphs. Original western blots can be found at Supplementary Materials.

Article Snippet: MEK inhibitor PD98059 (10 μM) and JNK inhibitor SP600125 (10 μM) were purchased from Selleckchem (Munich, Germany) and treated for the indicated times.

Techniques: Activation Assay, Activity Assay, Transfection, Construct, Luciferase, Expressing, Quantitative RT-PCR, Control, Western Blot