Antioxidant Activity Mediates Pirfenidone Antifibrotic Effects in Human Pulmonary Vascular Smooth Muscle Cells Exposed to Sera of Idiopathic Pulmonary Fibrosis Patients

Figure 3

(a-b) Effects of IPF sera on HPASMC collagen I production. (a) Before stimulation, subconfluent HPASMCs were transduced with lentiviral particles obtained from the COL1A1-LV-tGFP and EF1α-LV-FP602 lentivectors and then cultured in basal medium containing 10% (v/v) of sera from idiopathic pulmonary fibrosis (IPF), sera from idiopathic pulmonary fibrosis patients treated for 24 weeks with Pirfenidone (IPF + D), and healthy donors (HD). Data represent the collagen I promoter activity after 8 hours of sera stimulation. Data are normalized for transduction efficiency by reporting the ratio of COL1A1-LV-tGFP to EF1α-LV-FP602 relative fluorescence units (RFUs). (b) Subconfluent HPASMCs were stimulated for 48 hrs with basal medium containing 10% (v/v) of sera from idiopathic pulmonary fibrosis (IPF), sera from idiopathic pulmonary fibrosis patients treated for 24 weeks with Pirfenidone (IPF + D), and healthy donors (HD) and processed for collagen I quantification as reported in the Materials and Methods. In selected experiments, cells were pretreated for 60 min with the NADPH oxidase inhibitor diphenyleneiodonium (DPI) before exposure to the sera. Data are expressed as ng/ml collagen protein. value indicating that the significance is reported in the figure.

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