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

Figure 2

(a-b) Effects of IPF sera on HPASMC intracellular ROS levels. Before stimulation, subconfluent human pulmonary artery smooth muscle cells (HPASMCs) were loaded with 10 μM of H₂-DCFDA, 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). (b) In selected experiments, cells were pretreated for 60 min with the NADPH oxidase inhibitor diphenyleneiodonium (DPI) before exposure to the sera. (a-b) Data represent the variations of intracellular ROS levels after 2 hours of sera stimulation. Fluorescence data were normalized for protein content and expressed as relative fluorescence units (RFUs). value indicating that the significance is reported in the figure.

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