College of Life Sciences and Key Laboratory of Bioactive Materials Ministry of Education, Nankai University, Tianjin, P. R. China.
WuQing TCM Hospital Affiliated to Tianjin University of Traditional Chinese Medicine, Tianjin, P. R. China.
Nat Prod Res. 2020 May;34(10):1437-1441. doi: 10.1080/14786419.2018.1512994. Epub 2018 Nov 16.
In the study, we treated C6 rat glioma cells with 25 mg/ml Dulcitol for 24 h. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays were used to detect cellular growth. The measurements of the superoxide dismutase (SOD), malondialdehyde (MDA) and catalase (CAT) were used to assess oxidative stress level. Western was performed to detect the autophagy and apoptosis expression. The data showed that Dulcitol significantly decreased the cell viability, upregulated the Bax level in mitochondria and the Cytochrome C level in cytoplasm, and downregulated anti-apoptotic protein Bcl-xl. Moreover, it enhanced MDA level, reduced CAT and SOD activities, decreased LC3-II/LC3-I ratio, and increased P62 expression. However, rapamycin increased autophagy level and cell viability, and decreased ROS in Dulcitol treated C6 cells. Moreover, Dulcitol inhibited the glioma growth and enhanced survival . These results suggest that Dulcitol evidently increase cellular ROS levels and apoptosis in glioma cells, which can be significantly regulated by autophagy.
在这项研究中，我们用 25mg/ml 山梨醇处理 C6 大鼠神经胶质瘤细胞 24 小时。采用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐 (MTT) 法检测细胞生长情况。通过测定超氧化物歧化酶 (SOD)、丙二醛 (MDA) 和过氧化氢酶 (CAT) 的水平来评估氧化应激水平。采用 Western blot 法检测自噬和凋亡表达。结果表明，山梨醇显著降低细胞活力，上调线粒体中 Bax 水平和细胞质中细胞色素 C 水平，并下调抗凋亡蛋白 Bcl-xl。此外，它还增加 MDA 水平，降低 CAT 和 SOD 活性，降低 LC3-II/LC3-I 比值，增加 P62 表达。然而，雷帕霉素增加了自噬水平和细胞活力，降低了山梨醇处理的 C6 细胞中的 ROS。此外，山梨醇抑制了胶质瘤的生长并提高了生存能力。这些结果表明，山梨醇明显增加了胶质瘤细胞中的 ROS 水平和细胞凋亡，而自噬可以显著调节这些作用。
