目的 基于 SIRT1/PGC-1α 通路探讨加味孔圣枕中丹含药血清对氧糖剥夺再灌注（OGD/R）损伤 PC12 细 胞的线粒体功能的改善作用及机制。方法 采用大鼠 PC12 细胞构建体外 OGD/R 细胞模型。细胞分组：正常 组（10% FBS）、模型组（10% FBS）、10% 含药血清组、5% 含药血清组（5% 含药血清+5% 空白血清）、10% 空 白血清组（对照组）。采用 CCK-8 法检测细胞存活率，筛选合适的氧糖剥夺时间（2、4、6、8 h）；MTT 法检测 细胞活性；线粒体压力测试（MST）法检测细胞氧气消耗速率（OCR）；流式细胞术（Annexin V-PE/7-AAD 双染 法）检测细胞凋亡情况；Western Blot 法检测 PC12 细胞 SIRT1、PGC-1α 蛋白表达水平。结果 与正常组比较， 氧糖剥夺 2、4、6、8 h 后的 PC12 细胞活性均明显下降（P＜0.05），选择氧糖剥夺 6 h 作为后续实验造模时间。 与正常组比较，模型组的细胞活性明显下降（P＜0.05）；细胞基础呼吸值、最大呼吸值、质子漏、ATP 产生、 备用呼吸能力的 OCR 值均明显降低（P＜0.05）；细胞凋亡率明显升高（P＜0.05）；细胞中 SIRT1、PGC-1α 蛋白 表达水平明显下降（P＜0.05）。与模型组比较，加味孔圣枕中丹 10%、5% 含药血清组的细胞活性明显提高 （P＜0.05）；细胞基础呼吸值、最大呼吸值、质子漏、ATP 产生、备用呼吸能力的 OCR 值均明显升高（P＜ 0.05）；细胞凋亡率明显降低（P＜0.05）；细胞中 SIRT1、PGC-1α 蛋白表达水平明显升高（P＜0.05）。结论 加 味孔圣枕中丹含药血清能改善 OGD/R 损伤 PC12 细胞的线粒体功能障碍，抑制神经元凋亡，促进神经元存活， 其作用机制可能与激活 SIRT1/PGC-1α 信号通路有关。

Objective To investigate the improvement effect and mechanism of Modified Kongsheng Zhenzhong Dan medicated serum on mitochondrial function of PC12 cells injured by oxygen-glucose deprivation/reperfusion （OGD/R） based on the SIRT1/PGC-1α pathway. Methods PC12 cells were used to construct OGD/R cell model in vitro. Cell grouping：normal group （10% FBS），model group （10% FBS），10% drug-containing serum group，5% drugcontaining serum group （5% drug-containing serum + 5% blank serum），10% blank serum group （control group）. CCK-8 method was used to detect the cell survival rate，and the appropriate oxygen glucose deprivation time （2，4，6，8 hours） was screened. MTT assay was used to detect cell activity；mitochondrial stress test （MST） was used to detect the oxygen consumption rate （OCR）. Apoptosis was detected by flow cytometry （Annexin V-PE/7-AAD double staining）. The protein expression levels of SIRT1 and PGC-1α in PC12 cells were detected by Western Blot. Results Compared with the normal group， the activity of PC12 cells decreased significantly after oxygen-glucose deprivation for 2，4，6 and 8 hours （P＜0.05），and 6 hours of oxygen-glucose deprivation was selected as the time of subsequent experimental modeling. Compared with the normal group，the cell activity of the model group was significantly decreased （P＜0.05） . The OCR values of basic respiration value，maximum respiration value，proton leakage，ATP production and standby respiration ability were significantly decreased （P＜0.05）. The apoptosis rate was significantly increased （P＜0.05）. The protein expression levels of SIRT1 and PGC-1α in cells were significantly decreased （P＜0.05）. Compared with the model group， the cell activity of the Modified Kongsheng Zhenzhong Dan 10% and 5% drug-containing serum groups was significantly increased （P＜0.05） . The OCR values of basal respiration value，maximum respiration value，proton leakage，ATP production and spare breathing ability were significantly increased （P＜0.05）. The apoptosis rate was significantly decreased （P＜0.05）. The protein expression levels of SIRT1 and PGC-1α in cells were significantly increased （P＜0.05）. Conclusion The Modified Kongsheng Zhenzhong Dan medicated serum can improve mitochondrial dysfunction， inhibit neuronal apoptosis and promote neuronal survival in OGD/R-injured PC12 cells. The mechanism may be related to the activation of SIRT1/PGC-1α signaling pathway.

R285.5

国家自然科学基金项目（81202641）；山东省自然科学基金项目（ZR2020MH345）；济南市高校自主创新团队项目（2020GXRC012）