Figure 5

LPS induced EC dysfunction and collagen I mRNA synthesis. Cultured ECs were incubated with LPS, 2 μg/ml, or LPS, 4 μg/ml, for 24 hours. (A) EC viability was evaluated with FACS analysis (AnnV/PI). Only a small percentage of ECs underwent apoptosis after LPS, 2 μg/ml (8.9% ±1.6 versus basal 5. 9% ±1.2) and LPS, 4 μg/ml (7.8% ±1.6) stimulation compared with ECs treated with H₂O₂ 100 μM for 24 hours (positive control). (B) MTT cell-viability assay highlighted a significant proliferation of ECs after LPS, 2 μg/ml (0.720 ± 0.03 versus basal 24-hour 0.469 ± 0.004; P = 0.03) and LPS 4 μg/ml (0.597 ± 0.03 versus basal 24 hour 0.469 ± 0.004, P = 0.04) stimulation. (C, D) FACS analysis of EC showed the phenotypic changes induced by 24 hours of LPS stimulations. Results are expressed as mean ± SD and are representative of three independent experiments. (E) Real-time RT-PCR revealed the mRNA expression levels of collagen I. The gene relative expression was normalized to the expression of GAPDH. The histograms represent the mean ± SEM and are representative of three independent experiments.