丹酚酸B 通过mTOR / Ulk1 信号通路诱导胶质瘤细胞C6 自噬性死亡 |
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引用本文: | 赵敏,张瑜,张岚,刘欣,王新成.丹酚酸B 通过mTOR / Ulk1 信号通路诱导胶质瘤细胞C6 自噬性死亡[J].中国免疫学杂志,2018,34(4):549. |
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作者姓名: | 赵敏 张瑜 张岚 刘欣 王新成 |
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作者单位: | 郑州大学附属郑州中心医院神经内科;河南省直第三人民医院急诊科;郑州大学附属郑州中心医院高新区内四科 |
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摘 要: | 目的:探究丹酚酸B(Salvianolic acid B,Sal B)对胶质瘤细胞自噬的作用机制。方法:不同浓度的Sal B 处理细胞,CCK8 检测细胞活性,流式检测细胞凋亡情况,Western blot 检测增殖、凋亡及自噬标记蛋白表达,免疫荧光检测LC3 含量。结果:Sal B 浓度达到100 μmol/ L 时能明显抑制细胞活性,因此,选择10、20、50 μmol/ L 三个剂量进行后续试验;Sal B 能显著促进C6 细胞凋亡和凋亡标记蛋白caspase-3、Bax 的表达,降低Bcl-2 及增殖标记蛋白Ki67 的表达水平,并具有量效关系;同时,Sal B 能显著促进自噬标记蛋白(Atg5、Atg7、Atg12、Beclin1)的表达,升高LC3Ⅱ/ Ⅰ的比值,促进LC3 荧光斑点的形成并降低p62 的表达水平;此外,Sal B 可明显抑制mTOR/ Ulk1 通路蛋白mTOR 的表达,促进Ulk1 的表达。结论:Sal B 可通过mTOR/ Ulk1 信号通路诱导胶质瘤细胞自噬性死亡。
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关 键 词: | 丹酚酸B 胶质瘤 自噬 mTOR/ Ulk1 信号通路 |
Salvianolic acid B induces autophagic cell death in giloma through mTOR / Ulk1 signaling pathway |
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Abstract: | Objective:To evaluate the effects and underlying molecular mechanism of salvianolic acid B(Sal B) on autophagy in glioma cell.Methods:The C6 cells were treated with Sal B at a concentration ranging from 0 to 400 μmol/ L and the cell viability were measured by CCK8 assay.Flow cytometry was performed for apoptosis,Western blot assay for protein expression and the expression of LC3 was calculated by immunofluorescence.Results:The cell viability was weakened by Sal B at a concentration of 100 μmol/ L, thus,the concentration of 10,20 and 50 μmol/ L were selected for further experiment.Sal B dose-dependedly induced C6 cell apoptosis and the expression of caspase-3 and Bax,notably decreased the protein level of Bcl-2 and Ki67.Meanwhile,Sal B significantly up-regulated the expression level of Atg5,Atg7,Atg12 and Beclin1,increased the ratio of LC3Ⅱ/ Ⅰand the amount of LC3-labeled puncta per cell,and down-regulated the expression of p62.In addition,the protein level of mTOR was makedly decreased by Sal B and Sal B induced the expression of Ulk1.Conclusion:Sal B induces autophagic cell death in glioma cell through mTOR/ Ulk1 signaling pathway. |
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