The stimulatory effect of porcine placenta extract on in vitro high glucose-induced angiogenesis disturbance

Abstract

High glucose (HG)-induced reactive oxygen species (ROS) overproduction impairs angiogenesis that is one pivotal factor of wound healing process. Angiogenesis impairment induces delayed wound healing, whereby it has a higher risk of amputation in poorly controlled diabetic patient with diabetic foot ulcers. Porcine placenta extract (PPE) is a natural waste product that comprises a plenty of bioactive agents including growth factors and antioxidants. It was reported as an effective compound that prevents ROS generation. The goal of this study was to investigate the in vitro stimulatory effect of PPE on HG-induced angiogenesis disturbance. The primary endothelial cells (HUVECs) and endothelial cell line (EA.hy926) were treated with HG in the presence of PPE. The endothelial cells (ECs) viability, intracellular ROS generation, migration, angiogenesis, and apoptosis were determined by MTT assay, DCFDA reagent, wound healing assay, tube formation assay, and Annexin V/PI staining, respectively. Additionally, the molecular mechanism of PPE on HG-induced angiogenesis disturbance was investigated by Western blot analysis. The angiogenic growth factor secretion was also investigated by the sandwich ELISA technique. The results showed HG in the presence of PPE significantly decreased intracellular ROS overproduction compared to HG alone. HG in the presence of PPE significantly increased ECs viability, migration, and angiogenesis compared to HG alone by showing the recovery of PI3K/Akt/ERK1/2 activation. HG in the presence of PPE also decreased ECs apoptosis compared to HG alone by decreasing p53, Bax, cleaved caspase 9, cleaved caspase 3 levels and increasing Bcl 2 level. In summary, PPE attenuated HG-induced intracellular ROS overproduction and improved ECs viability, proliferation, migration, and angiogenesis by showing the recovery of PI3K/Akt/ERK1/2 activation and inhibition of ECs apoptosis. This study suggests PPE ameliorated HG-induced ROS-mediated angiogenesis impairment, whereby it potentially provides an alternative treatment for diabetic wounds.

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