目的：探讨芝麻酚（SEM）通过腺苷酸活化蛋白激酶（AMPK）/沉默信息调节因子1（SIRT1）/核因子κB（NF-κB）通路影响食管鳞状细胞癌（ESCC）Eca109 细胞自噬和凋亡的机制。方法: 用不同浓度的SEM（0、1.562 5、3.125、6.25、12.5、25、50、100、200、400 μmol/L）分别处理Eca109 细胞、人食管上皮细胞HEEpiC 48 h，CCK-8法检测细胞增殖率，筛选适宜的SEM浓度用于后续实验。将Eca109 细胞分为对照组（CK组，0 μmol/L）、低剂量SEM组（SEM-L 组，25 μmol/L）、中剂量SEM组（SEM-M 组，50 μmol/L）、高剂量SEM 组（SEM-H 组，100 μmol/L）、高剂量SEM+Compound C（AMPK抑制剂）组（SEM-H+Compound C组，100 μmol/L+10 μmol/L），所有各组Eca109 细胞在对应的药物浓度下处理48 h后，CCK-8法检测Eca109 细胞增殖，流式细胞术检测细胞凋亡，透射电镜观察Eca109 细胞内自噬小体，WB法检测Eca109 细胞中微管相关蛋白1轻链3（LC3）-Ⅱ/LC3-Ⅰ、Beclin-1、B淋巴细胞瘤2（Bcl2）、Bcl2相关X蛋白（BAX）、p-AMPK、SIRT1、p-NF-κB p65的表达。结果: 通过预实验选择SEM 实验浓度为25、50、100 μmol/L 用于正式研究。在SEM处理下，与CK组比较，SEM-L组、SEM-M 组、SEM-H 组Eca109 细胞的增殖水平（24、48 h）和Bcl2、p-NF-κB p65蛋白表达均显著降低，细胞凋亡率和自噬小体数量、LC3-Ⅱ/LC3-Ⅰ、Beclin-1、BAX、p-AMPK、SIRT1蛋白表达显著升高，且呈剂量依赖性（均P<0.05）；与SEM-H 组比较，SEM-H+Compound C 组 Eca109 细胞增殖水平（24、48 h）和Bcl2、p-NF-κB p65 蛋白表达均显著升高，细胞凋亡率和自噬小体数量、LC3-Ⅱ/LC3-Ⅰ、Beclin-1、BAX、p-AMPK、SIRT1蛋白表达均显著降低（均P<0.05）。结论:SEM可能通过激活AMPK/SIRT1信号通路而抑制NF-κB活性来促进Eca109细胞自噬与凋亡。

Objective: To explore the effect of sesamol (SEM) on autophagy and apoptosis of esophageal squamous cell carcinoma (ESCC) Eca109 cells through adenylate activated protein kinase (AMPK)/silencing information regulator 1 (SIRT1)/nuclear factor κB （NF- κB) pathway. Methods: Eca109 cells of ESCC and HEEpiC of human esophageal epithelial cells were treated with different concentrations of SEM (0, 1.562 5, 3.125, 6.25, 12.5, 25, 50, 100, 200, and 400 μmol/L) for 48 hours. Cell viability was detected by CCK-8 method and appropriate SEM concentrations were screened for subsequent experiments. Eca109 cells were grouped into control group (CK group, 0 μmol/L), low-dose SEM group (SEM-L group, 25 μmol/L), medium-dose SEM group (SEM-M group, 50 μmol/L),high-dose SEM group (SEM-M group, 50 μmol/L), high-dose SEM group (SEM-H group, 100 μmol/L) and high-dose SEM+compound C (AMPK inhibitor) group (SEM-H+Compound C group, 100 μmol/L+10 μmol/L). All Eca109 cells were treated at the corresponding drug concentration for 48 hours. CCK-8 assay was applied to detect the proliferation of Eca109 cells; flow cytometry was applied to detect apoptosis; transmission electron microscopy was applied to observe autophagosomes in Eca109 cells and WB assay was applied to detect the protein expressions of microtubule-associated protein 1 light chain 3 (LC3)-Ⅱ/LC3-Ⅰ, Beclin-1, B-lymphoma-2 (Bcl2),Bcl2-associated X protein (BAX), p-AMPK, SIRT1, p-NF-κB p65 in Eca109 cells 2. Results: The SEM concentrations of 25, 50 and 100 μmol/L selected through pre-experiment are used for formal research. Under SEM processing, compared with the CK group, the proliferation levels (24, 48 h) of Eca109 cells, protein expressions of Bcl2 and p-NF-κB p65 in SEM-L group, SEM-M group and SEM-H group decreased significantly while the apoptosis rate, the number of autophagosomes, the protein expressions of LC3Ⅱ-/LC3-Ⅰ,Beclin-1, BAX, p-AMPK and SIRT1 increased significantly, in a dose-dependent manner (all P<0.05); compared with the SEM-H group, the proliferation level (24, 48 h) of Eca109 cells, protein expressions of Bcl2 and p-NF-κB p65 in SEM-H+Compound C group increased significantly while the apoptosis rate, the number of autophagosomes, the protein expressions of LC3Ⅱ-/LC3-Ⅰ, Beclin-1,BAX, p-AMPK and SIRT1 decreased significantly(all P<0.05). Conclusion: SEM may promote autophagy and apoptosis of Eca109 cells by activating AMPK/SIRT1 signaling pathway and inhibiting NF-κB vitality.

湖北省科技厅科研计划项目（No.2019CFB789)