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目的探讨喹啉酸(QA)是否引起大鼠肾上腺嗜铬细胞瘤PC12细胞自噬的发生及其与糖原合酶激酶-3β(GSK-3β)/β-联蛋白相关信号通路的关系。方法用不同浓度QA(2.5,5和10 mmol·L~(-1))处理PC12细胞24 h,MTT法测定细胞存活,免疫荧光法观察微管相关蛋白1轻链3(LC3)荧光斑点标记的自噬体形成,Western蛋白印迹法检测细胞GSK-3β、β-联蛋白、LC3和Beclin 1的表达。结果 QA作用24 h,可浓度依赖性地降低PC12细胞存活率,IC_(50)为8.7 mmol·L~(-1)。QA作用24 h,与正常对照组比较,PC12细胞内LC3荧光斑点标记的自噬体增多,自噬相关蛋白LC3-Ⅱ/LC3-Ⅰ的比值和Beclin 1表达增高(P<0.01);同时PC12细胞内GSK-3β的表达增加(P<0.05,P<0.01),β-联蛋白的表达降低(P<0.05,P<0.01)。结论 QA可诱导PC12细胞自噬的发生,其机制可能与GSK-3β/β-联蛋白相关的信号通路激活有关。
Objective To investigate whether quinolinic acid (QA) causes autophagy in PC12 cells of adrenal pheochromocytoma and its relationship with glycogen synthase kinase-3β (GSK-3β) / β-catenin signaling pathways. Methods PC12 cells were treated with different concentrations of QA (2.5, 5 and 10 mmol·L -1) for 24 h. Cell viability was determined by MTT assay. Fluorescent labeling of microtubule-associated protein 1 light chain 3 (LC3) Of autophagosomes. Western blotting was used to detect the expression of GSK-3β, β-catenin, LC3 and Beclin 1. Results QA could decrease the survival rate of PC12 cells in a concentration-dependent manner for 24 h, with an IC 50 value of 8.7 mmol·L -1. Compared with the normal control group, the number of LC3-labeled autophagosomes increased, the ratio of autophagy-related protein LC3-Ⅱ / LC3-Ⅰ and the expression of Beclin 1 increased (P <0.01), while PC12 The intracellular GSK-3β expression increased (P <0.05, P <0.01), and the expression of β-catenin decreased (P <0.05, P <0.01). Conclusion QA can induce the autophagy in PC12 cells, which may be related to the activation of GSK-3β / β-catenin signaling pathway.