Periplaneta americana peptide decreases apoptosis of pig‐ovary granulosa cells induced by H₂O₂ through FoxO1

Abstract

Oxidative stress can induce apoptosis of granulosa cells and lead to follicular atresia, thereby reducing the number of pigs giving birth. The aim of this study was to investigate the protective effect of Periplaneta americana peptide (PAP) on the apoptosis of the granulosa cells of pig ovaries (PGCs) induced by hydrogen peroxide (H 2 O 2 ) via FoxO1. PGCs were treated with H 2 O 2 to establish a cell apoptosis model. Cell viability was measured using the cell counting kit-8 (CCK-8) assay, and cell apoptosis was detected using flow cytometry. The malondialdehyde (MDA) level and nitric oxide (NO) content were detected to reflect the oxidative stress. Western blotting, qRT-PCR and overexpression were undertaken to determine the expression of FoxO1 and caspase-3, and immunofluorescence was used to detect FoxO1 in the nucleus and cytoplasm. PGCs were treated with 100 μM H 2 O 2 for 6 hr, which resulted in oxidative damage and apoptosis and an apoptosis rate for PGCs of 32.95%. Next, PGCs were treated with 400 μg/ml PAP for 24 hr to repair the apoptosis induced by H 2 O 2 . PAP improved cell viability in H 2 O 2 -stimulated PGCs, the increased MDA level and NO content caused by H 2 O 2 stimulation were reversed and the apoptotic rate of PGCs was reduced. The qRT-PCR and Western blotting results indicated that PAP decreased the H 2 O 2 -induced apoptosis and the expression of FoxO1 and caspase-3 in PGCs. The effect of PAP was the same following FoxO1 overexpression. FoxO1 was expressed in the nucleus when stimulated by H 2 O 2 or overexpression; however, it migrated to the cytoplasm following PAP treatment. PAP decreased the apoptosis of PGCs induced by H 2 O 2 by regulating FoxO1 expression and nuclear translocation.

Mechanisms of and Potential Medications for Oxidative Stress in Ovarian Granulosa Cells: A Review

“…Modern medicine has further studied Periplaneta americana extract and found that it possesses several outstanding biological properties, including antibacterial, antioxidant, anti-inflammatory, and anti-cancer properties [129,130]. It has been demonstrated that Periplaneta americana peptide can significantly inhibit H 2 O 2 -induced apoptosis in KGN cells and porcine ovarian GCs by upregulating SIRT1 expression and reducing FOXO1 expression [73,131,132]. Moreover, Periplaneta americana peptide can also inhibit mitochondrial damage by upregulating Bcl2L13-mediated mitochondrial autophagy, thereby suppressing OS-induced apoptosis in KGN cells [107].…”