Abstract:Pumpkin seed oil prepared using continuous phase transition extraction, supercritical fluid extraction, and screw pressing was used to explore the in vivo inhibitory effect of pumpkin seed oil prepared using various extraction methods on BPH-1 cells. The cell survival rate, scratch healing rate, reactive oxygen species (ROS), malondialdehyde (MDA), tumor necrosis factor-α (TNF-α), and nitric oxide (NO) content were measured, and the expression level of the PI3K/Akt/NF-κB signaling pathway was analyzed. The results showed that the pumpkin seed oil prepared using the three extraction methods inhibited the survival rate and scratch healing rate of BPH-1 cells in a dose-dependent manner. Additionally, they reduced the secretion of oxidative stress indicators ROS and MDA, and the release of inflammatory factors TNF-α and NO, and down-regulated the expression of PI3K, Akt, and NF-κB mRNA. Among them, the relative content of ROS in BPH-1 cells of the pumpkin seed oil group prepared using continuous phase transition extraction at 75 μg•mL-1 mass concentration was 68.56%, the MDA content was 0.48 nmol•(mg prot)-1, the release of TNF-α was 60.99 pg•mL-1, and the release of NO was 4.32 μmol•L-1. These values were 31.44%, 85.92%, 78.04%, and 71.49% lower than those in the control group, respectively, and were significantly lower than the pumpkin seed oil groups prepared using supercritical fluid extraction and screw pressing (P<0.05). In summary, pumpkin seed oil has good anti-prostatic hyperplasia activity in vitro. The mechanism of this effect is associated with the inhibition of BPH-1 cell proliferation and migration, the ameliorative effect on oxidative stress and inflammatory levels, and the downregulation of the expression of the PI3K/Akt/NF-κB signaling pathway. The results contribute to a comprehensive understanding of the anti-prostatic hyperplasia activity of pumpkin seed oil and its mechanism, providing a theoretical basis for the development of related functional foods.
