为探究不同提取方法制备的南瓜籽油对BPH-1细胞的体外抑制作用，该研究以连续相变萃取技术、超临界流体萃取技术和螺旋压榨法制备的南瓜籽油为样品，测定细胞存活率、划痕愈合率、活性氧（ROS）、丙二醛（MDA）、肿瘤坏死因子-α（TNF-α）和一氧化氮（NO）含量，以及分析PI3K/Akt/NF-κB信号通路表达水平。结果显示，三种提取方法制备的南瓜籽油呈剂量依赖性抑制BPH-1细胞存活率和划痕愈合率，降低氧化应激指标ROS和MDA的分泌量以及炎症因子TNF-α和NO的释放量，同时下调PI3K、Akt、NF-κB mRNA的表达量。其中，75 μg?mL-1质量浓度下连续相变萃取技术制备的南瓜籽油组BPH-1细胞中的ROS相对含量为68.56%、MDA含量为0.48 nmol?mgprot-1、TNF-α释放量为60.99 pg?mL-1、NO释放量为4.32 μmol?L-1，相比于空白组分别降低了31.44%、85.92%、78.04%、71.49%，同时显著低于超临界流体萃取技术和螺旋压榨法制备的南瓜籽油组（P<0.05）。综上，南瓜籽油具有较好的体外抗前列腺增生活性，其作用机制与抑制BPH-1细胞的增殖和迁移，改善氧化应激和炎症水平，以及下调PI3K/Akt/NF-κB信号通路表达有关，该研究结果有助于全面理解南瓜籽油抗前列腺增生活性与机制，为开发相关功能性食品提供理论基础。

In order to explore the inhibitory effect of pumpkin seed oil prepared by different extraction methods on BPH-1 cells in vitro, the pumpkin seed oil prepared by continuous phase transition extraction, supercritical fluid extraction and screw pressing were used as samples. 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 by the three extraction methods inhibited the survival rate and scratch healing rate of BPH-1 cells in a dose-dependent manner, 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 pumpkin seed oil group prepared by continuous phase transition extraction at 75 μg?mL-1 mass concentration was 68.56%, the content of MDA was 0.48 nmol?mgprot-1, the release of TNF-α was 60.99 pg?mL-1, and the release of NO was 4.32 μmol?L-1. Compared with the blank group, they were reduced by 31.44%, 85.92%, 78.04%, and 71.49%, respectively, and were significantly lower than the pumpkin seed oil group prepared by supercritical fluid extraction and screw pressing (P<0.05). In summary, pumpkin seed oil has good anti-prostatic hyperplasia activity in vitro, and its mechanism is related to inhibiting the proliferation and migration of BPH-1 cells, improving oxidative stress and inflammatory levels, and down-regulating the expression of the PI3K/Akt/NF-κB signaling pathway. The results of this study are helpful to fully understand the anti-prostatic hyperplasia activity and mechanism of pumpkin seed oil, and provide a theoretical basis for the development of related functional foods.

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