目的： 研究c-Jun氨基末端激酶（c-Jun N-terminal kinase，JNK）在维生素E琥珀酸酯（vitamin E succinate，VES）激活内质网应激诱导人胃癌SGC-7901细胞自噬过程中发挥的作用。方法： 用不同剂量VES（5、10、15、20 μg/ml）分别处理SGC-7901细胞24 h，采用qPCR和WB法分别检测自噬标志物LC3和Beclin-1 mRNA和蛋白的表达水平；在内质网应激抑制剂4-PBA的作用下，激光共聚焦显微镜观察LC3的荧光强度和分布，然后采用qPCR检测内质网应激标志物GRP78、GRP94 mRNA和自噬标志物LC3、Beclin-1 mRNA的表达水平；在JNK抑制剂SP600125的作用下，采用WB法检测p-JNK和自噬标志蛋白LC3和Beclin-1的表达变化。结果： 与溶剂对照组相比，随着VES作用浓度的升高，SGC-7901细胞中Beclin-1和LC3的mRNA和蛋白表达水平呈现逐渐升高的趋势（均P<0.05），且LC3-Ⅱ/LC3-Ⅰ比值也显著增加（均P<0.01）；与20 μg/ml VES组比较，经内质网应激抑制剂4-PBA预处理组的SGC-7901细胞中GRP78、GRP94、LC3和Beclin-1 mRNA表达均降低（均P<0.01）、LC3的点状聚集强度下降；在JNK抑制剂SP600125预处理组的SGC-7901细胞中，p-JNK、LC3、Beclin-1的蛋白表达水平均较20 μg/ml VES组降低（均P<0.01）。结论： VES可通过激活内质网应激诱导SGC-7901细胞发生自噬，同时JNK参与了内质网应激对自噬的调控过程。

Objective: To study the role of c-Jun N-terminal kinase (JNK) in vitamin E succinate (VES) activating endoplasmic reticulum stress-induced autophagy in human gastric SGC-7901 cells.Methods: SGC-7901 cells were treated with different doses of VES (5, 10, 15, 20 μg/ml) for 24 h, then, qPCR and WB were used to detect the mRNA and protein expressions of autophagy markers LC3 and Beclin-1; Under the action of reticulum stress inhibitor 4-PBA, the fluorescence intensity and distribution of LC3 were observed under laser confocal microscope, and then qPCR was used to detect the mRNA expression of endoplasmic reticulum stress markers GRP78, GRP94 and autophagy markers LC3, Beclin-1. Under the action of the JNK inhibitor SP600125, WB was used to detect the protein expression changes of p-JNK and autophagy marker proteins LC3 and Beclin-1.Results: Compared with the control group, with the increase of the concentration of VES, the mRNA and protein expressions of Beclin-1 and LC3 showed a gradual increase (all P < 0.05), and LC3-Ⅱ/LC3-Ⅰ ratio also increased significantly (P < 0.01); compared with 20 μg/ml VES group, after pretreatment with endoplasmic reticulum stress inhibitor 4-PBA, mRNA expressions of GRP78, GRP94, LC3 and Beclin-1 decreased (all P < 0.01), and the intensity of LC3 punctate aggregation decreased; after pretreatment with JNK inhibitor SP600125, the protein expression levels of p-JNK, LC3 and Beclin-1 were lower than those of 20 μg/ml VES group (all P < 0.01), these results indicated that inhibition of JNK activity could inhibit the occurrence of autophagy.Conclusion: VES can induce autophagy in SGC-7901cells by activating endoplasmic reticulum stress, and JNK participates in the regulation process of endoplasmic reticulum stress on autophagy.

R735.2；R730.54

河北省自然科学基金青年科学基金资助项目（No. H2019209453）；华北理工大学博士启动基金资助项目（No. 28409599）