Abstract
The mechanism of action by which oxidative stress induces granulosa cell apoptosis, which plays a vital role in initiating follicular atresia, is not well understood. In the present study, the effect of 3-nitropropionic acid (3-NPA) on oxidative stress and apoptosis in granulosa cells in geese was investigated. Our results showed that treatment with 3-NPA at 5.0 mmol/l for 24 h increased intracellular reactive oxygen species (ROS) production by 25.4% and decreased granulosa cell viability by 45.5% (P<0.05). Catalase and glutathione peroxidase gene expression levels in granulosa cells treated with 3-NPA were 1.32- and 0.49-fold compared with those of the control cells, respectively (P <0.05). A significant decrease in the expression level of B-cell lymphoma 2 (Bcl-2) protein and remarkable increases in the levels of Bax, p53 and cleaved-Caspase 3 proteins and the ratio of Bax/Bcl-2 expression in granulosa cells treated with 3-NPA were observed (P<0.05). Furthermore, a 38.43% increase in the percentage of early apoptotic cells was also observed in granulosa cells treated with 3-NPA (P<0.05). Moreover, the expression levels of NF-κB, Nrf2, Fhc, Hspa2 and Ho-1 in granulosa cells treated with 3-NPA were elevated 4.36-, 1.63-, 3.62-, 27.54- and 10.48-fold compared with those of the control cells (P<0.05), respectively. In conclusion, the present study demonstrates that treatment with 3-NPA induces ROS production and apoptosis and inhibits the viability of granulosa cells in geese. Furthermore, 3-NPA triggers increases in the expression of cleaved-Caspase 3 protein and the ratio of Bax/Bcl-2 expression, and induces the early apoptosis of granulosa cells.
Highlights
Endogenous reactive oxygen species (ROS) play vital roles in signaling pathways including cell growth, metabolism, differentiation and apoptosis [1,2]
No significant difference in Sod expression level was observed. These results indicated that ROS products induced by 3-nitropropionic acid (3-NPA) modulated the levels of Cat and Gpx mRNA expression in granulosa cells
Fhc mRNA expression up-regulation by 3-NPA indicated that Ferritin heavy chain (FHC) might be essential for modulating the response of granulosa cells to oxidative stress induced by 3-NPA through the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and nuclear factor (erythroid-derived 2)-like 2 (Nrf2) antioxidant pathways, the exact mechanism remains to be investigated
Summary
Endogenous reactive oxygen species (ROS) play vital roles in signaling pathways including cell growth, metabolism, differentiation and apoptosis [1,2]. Excess ROS production causes lipid peroxidation and DNA damage, thereby inducing cell death [3,4]. Several studies have suggested that numerous ROS are produced by the ovaries in female animals during folliculogenesis and ovulation [5,6]. Granulosa cells, which surround oocytes in ovaries, play a pivotal role in follicular development, ovulation, atresia and steroidogenesis [7,8]. It is well known that oxidative stress induces granulosa cell apoptosis, which plays a vital role in initiating follicular atresia [9,10,11]. An understanding of these mechanisms is limited and remains to be defined
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