目的 明确当归通过调节鞘脂代谢发挥对皮质酮（CORT）损伤的PC12细胞的保护作用及机制。方法 制备当归水提物，LC-MS法进行成分鉴定；当归水提物含药血清（ASWEMS）的制备：将12只SPF级雄性SD大鼠随机分为空白血清（给予0.9%氯化钠溶液）组和ASWEMS（生药量7.5 g·kg^-1）组，连续ig给药6 d，采血管腹主动脉采血，分离血清后同组混合，56℃水浴加热30 min后经0.22 μm微孔滤膜滤过除菌。利用CORT诱导PC12细胞构建损伤模型，将PC12细胞分为：对照（5%空白血清）组、模型组（400 μmol·L^-1 CORT＋5%空白血清）、ASWEMS组（400 μmol·L^-1 CORT＋5% ASWEMS）、神经酰胺合成酶抑制剂伏马菌素（FB1，400 μmol·L^-1 CORT＋5%空白血清＋10 μmol·L^-1 FB1）组，处理24 h。通过CCK-8法检测细胞存活率；采用Annexin FITC/PI流式细胞术检测细胞凋亡率；基于超高效液相色谱-四极杆-飞行时间质谱（UPLC-Q-TOF-MS）脂质组学技术检测PC12细胞脂质代谢轮廓变化及鞘脂类代谢物相对含量；采用ELISA检测PC12细胞神经酰胺（Cer）含量。结果 LC-MS法鉴定出当归水提物17种化学成分。与模型组比较，ASWEMS和FB1均可显著回调CORT损伤PC12细胞所引起的细胞存活率下降（P＜0.001），细胞凋亡率增加（P＜0.001）。鞘脂类代谢物是引起对照组与模型组差异贡献较大的代谢物，与对照组比较，CORT干预24 h后，PC12细胞中的二氢鞘氨醇（Dhsph）类、植物鞘氨醇（PhytoSph）类代谢物相对含量显著降低（P＜0.001），鞘磷脂（SM）类代谢物相对含量显著升高（P＜0.001），Cer相对含量及绝对含量均显著升高（P＜0.01）；与模型组比较，给予ASWEMS、FB1干预后各代谢物含量具有不同程度的回调（P＜0.01、0.001）。结论 ASWEMS对CORT诱导的PC12细胞损伤具有保护作用，其机制与调节鞘脂代谢通路、抑制细胞凋亡有关。

Objective To clarify the protective effect of Angelica sinensis water extract medicated serum (ASWEMS) on PC12 cells damaged by corticosterone (CORT) through the regulation of sphingolipid metabolism. Methods Preparation of Angelica sinensis water extract and identification of its components by LC-MS method. Preparation of Angelica sinensis water extract medicated serum (ASWEMS): Twelve healthy male SD rats were randomly divided into blank serum (received 0.9% sodium chloride solution) group and ASWEMS (crude drug dose of 7.5 g·kg^-1) group, acclimated for one week, and then intragastrically administered ASWEMS for six consecutive days. Blood was collected from the abdominal aorta of the rats, and the serum was separated and mixed within the same group. The serum was then heated at 56 ℃ for 30 min and filtered through a 0.22 μm microfiltration membrane to remove bacteria. A cell injury model was constructed by treating PC12 cells with CORT, with the cells divided into control (5% control serum), model (400 μmol·L^-1 CORT＋5% control serum), ASWEMS (400 μmol·L^-1 CORT＋5% ASWEMS) and fumonisin (FB1, 400 μmol·L^-1 CORT＋5% control serum ＋10 μmol·L·L^-1 FB1) groups, which subjected to the treatment for 24 h. The cell survival rate was detected by CCK-8 method. Annexin FITC/PI flow cytometry was used to detect apoptosis rate. Changes in lipid metabolism profile and the relative content of sphingolipid metabolites in PC12 cells were determined by ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). The contents of Ceramide (Cer) in PC12 cells was detected by ELISA. Results LC-MS method identified 17 chemical components in Angelica sinensis water extract. Compared with the model group, ASWEMS and FB1 could significantly reverse the decreased cell viability and increased apoptosis rate caused by CORT injury in PC12 cells (P <0.001). Sphingolipids metabolites were the metabolites that contributed more to the difference between the control group and the model group. Compared with the control group, the relative content of Dhsph-type and PhytoSph-type metabolites in PC12 cells was significantly lower after CORT intervention for 24 h (P <0.001), while the relative content of SM-type metabolites was significantly higher (P <0.001), and the relative content and absolute content of Cer were both significantly higher (P <0.01). Compared with the model group, the contents of all metabolites were significantly adjusted after ASWEMS or FB1 intervention (P <0.01, 0.001). Conclusion ASWEMS has a protective effect against CORT-induced PC12 cell injury through regulating sphingolipid metabolism pathway and inhibiting apoptosis.

R285.5

国家自然科学基金项目（82004502）;山西省应用基础研究计划项目（202403021211187）;经方扶阳山西省重点实验室项目（CP‐SY202304）;山西省重点研发项目（202102130501010）