Bevacizumab Increases Endothelin-1 Production via Forkhead Box Protein O1 in Human Glomerular Microvascular Endothelial Cells In Vitro

Figure 4

Vascular endothelial growth factor A (VEGFA) is associated with the suppression of ET-1 production in cultured human glomerular microvascular endothelial cells (hGECs). hGECs were plated in 24-well plates (1 × 10⁵ cells/well) for ELISA or 60 mm culture dishes (1 × 10⁶ cells/dish) for RT-PCR. hGECs that were 90% confluent were serum-starved for 24 h before the experiments were performed. hGECs were treated with 0, 10, and 40 ng/mL of VEGFA or with a combination of 40 ng/mL VEGFA and 1 µM bevacizumab for 8 h. The levels of ET-1 mRNA (a) and ET-1 protein (b) were measured using real-time RT-PCR and the ELISA after VEGFA and bevacizumab treatment. Data shown are means ± SD (n = 3;  < 0.05). The experiments were repeated at least three times, with reproducible results.

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(b)