目的 利用皮质酮诱导大鼠肾上腺髓质嗜铬瘤分化细胞株PC12损伤，模拟糖皮质激素诱发神经功能障碍的病理过程，探讨沉香提取物对神经细胞的保护作用及其机制。方法 采用400 μmol/L皮质酮诱导的PC12细胞作为神经元损伤模型，给予沉香提取物（5、10、20 μg/mL）干预细胞24 h后，利用CCK-8试剂盒检测细胞存活率；采用Hoechst 33258染色和流式细胞术检测细胞凋亡率；通过Western blotting检测凋亡相关蛋白半胱氨酸天冬氨酸蛋白酶-3（cystein-asparate protease-3，Caspase-3）、cleaved Caspase-3、聚腺苷酸二磷酸核糖转移酶[poly （ADP-ribose） polymerase，PARP]、cleaved PARP表达情况。结果 与对照组比较，皮质酮处理PC12细胞24 h后细胞存活率显著降低（P<0.01），细胞凋亡率显著升高（P<0.01），cleaved PARP/PARP和cleaved Caspase-3/Caspase-3蛋白表达水平显著升高（P<0.01）。与模型组比较，给予沉香提取物24 h后细胞存活率显著升高（P<0.05、0.01），凋亡率显著降低（P<0.01），cleaved PARP/PARP和cleaved Caspase-3/Caspase-3蛋白表达水平显著降低（P<0.01）。结论 沉香提取物通过调控Caspase-3/PARP通路改善皮质酮诱导的PC12细胞凋亡，从而保护神经元。

Objective Corticosterone was used to induce the injury of rat adrenal medulla pheochromocytoma cell line PC12 to simulate the pathological process of glucocorticoid-induced neurological dysfunction, so as to explore the protective effect and mechanism of Chenxiang (Aquilariae Lignum Resinatum) extract on nerve cells. Methods PC12 cells induced by 400 μmol/L corticosterone were used as neuron injury model. After 24 h of intervention with Aquilariae Lignum Resinatum extract (5, 10, 20 μg/mL), cell survival rate was detected by CCK-8 kit; Hoechst 33258 staining and flow cytometry were used to detect cell apoptosis rate; Western blotting was used to detect apoptosis-related proteins cystein-asparate protease-3 (Caspase-3), cleaved Caspase-3, poly(ADP-ribose)polymerase (PARP) and cleaved PARP expressions. Results Compared with control group, survival rate of PC12 cells treated with corticosterone for 24 h was significantly decreased (P < 0.01), apoptosis rate was significantly increased (P < 0.01), cleaved PARP/PARP and cleaved Caspase-3/Caspase-3 protein expressions were significantly increased (P < 0.01). Compared with model group, survival rate of P12 cells treated with Aquilariae Lignum Resinatum extract for 24 h was significantly increased (P < 0.05, 0.01), apoptosis rate was significantly decreased (P < 0.01), cleaved PARP/PARP and cleaved Caspase-3/Caspase-3 protein expressions were significantly decreased (P < 0.01). Conclusion Aquilariae Lignum Resinatum extract improves corticosterone-induced apoptosis of PC12 cells by regulating PARP/Caspase-3 pathway, thereby protecting neurons.

R285.5

国家自然科学基金资助项目（82003912）；国家重点研发项目（2018YFC1706400）