目的 探究人参皂苷Re对鱼藤酮诱导的SH-SY5Y细胞帕金森病（Parkinson’s disease，PD）模型的保护作用机制。方法 利用0.3 μmol/L鱼藤酮建立SH-SY5Y细胞PD模型，给予2.5、5 μmol/L人参皂苷Re或10 μmol/L左旋多巴（levodopa，L-DOPA）处理24 h，通过CCK-8试剂盒检测细胞存活率；流式细胞仪检测细胞周期、凋亡、线粒体膜电位及活性氧（reactive oxygen species，ROS）含量变化；能量代谢分析仪检测细胞糖酵解功能及线粒体氧化磷酸化功能；ELISA检测细胞内三磷酸腺苷（adenosine triphosphate，ATP）含量变化；免疫荧光检测细胞内Parkin和PTEN诱导性激酶蛋白-1（PTEN induced putative kinase-1，Pink-1）的表达情况；Western blotting检测细胞内Parkin、Pink-1、细胞色素C（cytochrome C，Cyt-C）、B淋巴细胞瘤-2（B-cell lymphoma-2，Bcl-2）、Bcl-2相关X蛋白（Bcl-2 associated X protein，Bax）、半胱氨酸天冬氨酸蛋白酶-3（cystein-asparate protease-3，Caspase-3）、cleaved Caspase-3、核因子E2相关因子2（nuclear factor erythroid 2-related factor 2，Nrf2）、血红素氧合酶-1（heme oxygenase-1，HO-1）、谷氨酸半胱氨酸连接酶亚基（glutamate-cysteine ligase catalytic subunit，GCLC）和NADPH醌氧化还原酶-1（NADPH quinine oxidoreductase-1，NQO-1）蛋白表达的变化。结果 以0.3 μmol/L鱼藤酮建立SH-SY5Y细胞PD模型，细胞存活率显著下降（P＜0.01），细胞阻滞于G₀/G₁期，细胞凋亡比例显著升高（P＜0.01），线粒体膜电位及ATP含量显著下降（P＜0.01），ROS生成量显著升高（P＜0.01），糖酵解PER值、细胞的最大呼吸能力、备用呼吸能力及ATP合成能力均显著下降（P＜0.01），Parkin、Pink-1、Bcl-2、HO-1、GCLC、NQO1蛋白表达显著下调（P＜0.01），Cyt-C从线粒体向胞质大量释放（P＜0.01），Nrf2核转移降低（P＜0.01），Bax、cleaved Caspase-3蛋白表达显著上调（P＜0.01）；给予L-DOPA或人参皂苷Re干预后，明显增加细胞存活率（P＜0.05、0.01），降低细胞凋亡率（P＜0.01），减轻细胞G₀/G₁期阻滞作用（P＜0.05、0.01），增强细胞糖酵解PER值、细胞的最大呼吸能力、备用呼吸能力及ATP合成能力（P＜0.05、0.01），维护了线粒体功能稳态，表现为线粒体膜电位、ATP含量显著增加（P＜0.05、0.01），ROS生成量显著减少（P＜0.05、0.01），Parkin、Pink-1、Bcl-2、HO-1、GCLC、NQO1表达显著上调（P＜0.05、0.01），抑制Cyt-C从线粒体向细胞质释放（P＜0.05、0.01），Nrf2核转移增加（P＜0.05、0.01），Bax、cleaved Caspase-3表达显著下调（P＜0.05、0.01）。结论 人参皂苷Re可明显改善鱼藤酮诱导的细胞毒性，其机制可能是人参皂苷Re通过活化Parkin及Pink-1蛋白，维护线粒体功能稳态，激活Kelch样ECH相关蛋白1（Kelch like ECH associated protein 1，Keap1）-Nrf2-抗氧化反应元件（antioxidant response element，ARE）信号通路，抑制线粒体介导的Caspase级联反应，从而抑制神经细胞凋亡，发挥神经保护作用。

Objective To explore the protective mechanism of ginsenoside Re against rotenone-induced Parkinson’s disease (PD) model in SH-SY5Y cells. Methods PD model of SH-SY5Y cells was established induced by 0.3 μmol/L rotenone, cells were treated with 2.5, 5 μmol/L ginsenoside Re or 10 μmol/L levodopa (L-DOPA) for 24 h, cell survival rate was detected by CCK-8; The changes of cell cycle, apoptosis, mitochondrial membrane potential and reactive oxygen species (ROS) content were detected by flow cytometry; The cell glycolysis function and mitochondrial oxidative phosphorylation function were detected by energy metabolism analyzer; ELISA was used to detect the intracellular adenosine triphosphate (ATP) content changes; Immunofluorescence was used to detect intracellular expressions of Parkin and PTEN induced kinase protein-1 (PINK-1); Western blotting was used to detect intracellular Parkin, Pink-1, cytochrome C (Cyt-C), B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), cysteine aspartate protease-3 (Caspase-3), cleared Caspase-3, nuclear factor E2 related factor 2 (Nrf2), heme oxygenase-1 (HO-1), glutamate cysteine ligase catalytic subunit (GCLC) and NADPH quinine oxidoreductase-1 (NQO-1) expressions. Results PD model of SH-SY5Y cells was established with 0.3 μmol/L rotenone, cell survival rate was significantly decreased (P < 0.01), cells were arrested in G₀/G₁ phase, percentage of apoptosis was significantly increased (P < 0.01), mitochondrial membrane potential and ATP content were significantly decreased (P < 0.01), ROS production was significantly increased (P < 0.01), glycolytic PER value, maximum respiratory capacity, standby respiratory capacity and ATP synthesis capacity were significantly decreased (P < 0.01), Parkin, Pink-1, Bcl-2, HO-1, GCLC and NQO1 protein expressions were significantly decreased (P < 0.01), Cyt-C was released in large quantities from mitochondria to cytoplasm (P < 0.01), Nrf2 nuclear transfer was reduced (P < 0.01), Bax and cleaved Caspase-3 protein expressions were significantly upregulated (P < 0.01); After intervention with L-DOPA or ginsenoside Re, cell survival rate was significantly increased (P < 0.05, 0.01), cell apoptosis rate was reduced (P < 0.01), cell G₀/G₁ phase arrest was reduced (P < 0.05, 0.01), cell glycolysis PER value, cell maximum respiratory capacity, standby respiratory capacity and ATP synthesis capacity were enhanced (P < 0.05, 0.01), functional stability of mitochondria was maintained, manifested as mitochondrial membrane potential, ATP content were significantly increased (P < 0.05, 0.01), ROS production was significantly decreased (P < 0.05, 0.01), Parkin, Pink-1, Bcl-2, HO-1, GCLC and NQO1 expressions were significantly upregulated (P< 0.05, 0.01), Cyt-C release from mitochondria to cytoplasm was inhibited (P< 0.05, 0.01), Nrf2 nuclear transfer was increased (P < 0.05, 0.01), Bax and cleaved Caspase-3 expressions were significantly downregulated (P< 0.05, 0.01). Conclusion Ginsenoside Re can significantly improve the cytotoxicity induced by rotenone, and its mechanism may be that ginsenoside Re maintains mitochondrial functional homeostasis by activating Parkin and Pink-1 proteins, activating Kelch like ECH associated protein 1 (Keap1)-Nrf2-antioxidant response element (ARE) signaling pathway, and inhibiting mitochondrial mediated Caspase cascade reaction, thus inhibiting neuronal apoptosis and exerting neuroprotective effects.

山东省重大科技创新工程（2021CXGC010508）；山东省自然科学基金资助项目（ZR2021QH289）；国家重点研发计划（2019YFC1711205）；国家重点研发计划（2019YFC1711200）