Effect of suppressed PGC-1α expression on OA-induced VSMC proliferation and migration.

<p>VSMCs were transfected with siRNA S1 or the negative control (siRNA N). The interference effect was assessed by quantitative PCR (A) and western blot (B). Effects of decreased PGC-1α on OA-induced VSMC proliferation were determined by MTT assay (C) and cell counting (D). Effects of decreased PGC-1α on OA-induced VSMC migration were determined by wound healing (E) and transwell analysis (F). Data in VSMC proliferation and migration detections represent the means±SEM of 18 determinants from 3 independently prepared samples each with 6 measurements. Data in PGC-1α mRNA level detections are expressed as means±SEM of five different experiments normalized to β-actin levels. Data in PGC-1α protein level detections are expressed as means±SEM of four different experiments normalized to GAPDH levels. **P<0.01, #P<0.001 vs. control or N group.</p

Effects of PGC-1α on OA-induced p-ERK activity in VSMCs.

<p>PDGF-BB and pharmacological ERK-MAPK inhibitor PD98059 were chosen as positive and negative controls of ERK phosphorylation, respectively. VSMCs were made quiescent by serum-starvation for 24 h and then stimulated with PDGF-BB (100 ng/ml) for 30 min, PD98059 (50 uM) for 1 h before stimulation with PDGF-BB (100 ng/mL), or 0.4 mmol/L OA for 2 h. Proteins extracted following these treatments and phosphorylation of ERK was determined by western blot (A). VSMCs were also treated with 48 h adenovirus infection and then incubated with 0.4 mmol/L OA for 24 h. Phosphorylation of ERK was also analyzed with elevated PGC-1α level by western blot (B). Data were shown as the ratio of p-ERK/total ERK. The ratios of control were designated as 1.0. Individual data in this chart represent the mean±SEM of 4 determinants. #P<0.001 vs. control or positive/negative group.</p

Effect of overexpressed PGC-1α on OA-induced VSMC proliferation and migration.

<p>VSMCs were treated with 48 h adenovirus infection, then incubated with 0.4 mmol/L OA for 24 h. PGC-1α expression was analysised by quantitative PCR (A) and western blot (B). Effects of PGC-1α overexpression on OA-induced VSMC proliferation were determined by MTT assay (C) and cell counting (D). Effects of PGC-1α overexpression on OA-induced VSMC migration were determined by wound healing (E) and transwell analysis (F). Data in VSMC proliferation and migration detections represent the means±SEM of 18 determinants from 3 independently prepared samples each with 6 measurements. Data in PGC-1α mRNA level detections are expressed as means±SEM of five different experiments normalized to β-actin levels. Data in PGC-1α protein level detections are expressed as means±SEM of four different experiments normalized to GAPDH levels. *P<0.05, **P<0.01, #P<0.001 vs. control or GFP group.</p

Changes of PGC-1α expression and VSMC proliferation/migration in response to increased proportion of PA in fatty acid mixtures.

<p>VSMCs were incubated with various FFAs for 24 h before analysis. Effects of PA on PGC-1α expression was determined by quantitative PCR (A) and western blot analysis (B). Effects of increased PA on PGC-1α expression were determined by quantitative PCR (C). Effects of increased PA on OA-induced VSMC proliferation were determined by MTT assay (D) and cell counting (E). Effects of increased PA on OA-induced VSMC migration were determined by wound healing (F) and transwell analysis (G,H). Data in VSMC proliferation and migration detections represent the means±SEM of 18 determinants from 3 independently prepared samples each with 6 measurements. Data in PGC-1α mRNA level detections are expressed as means±SEM of five different experiments normalized to β-actin levels. Data in PGC-1α protein level detections are expressed as means±SEM of four different experiments normalized to GAPDH levels. *P<0.05, **P<0.01, #P<0.001 vs. control</p