丹参多酚酸盐诱导胶质瘤细胞自噬的实验研究  

The Experimental Research of Depsides Salts From Salvia Miltiorrhiza on Autophagy in Glioma Cell

作　　者：张小林[1] 张文静[1] 蒋峰[1] 刘晓光[1] 

机构地区：[1]温州医科大学附属舟山医院,浙江舟山316021

出　　处：《中华中医药学刊》2016年第4期972-975,I0012,共5页Chinese Archives of Traditional Chinese Medicine

基　　金：舟山市医药卫生科技计划项目(2012B14)

摘　　要：目的:研究丹参多酚酸盐对胶质瘤U251细胞自噬的影响。方法:丹参多酚酸盐作用胶质瘤U251细胞后,倒置显微镜动态观察细胞形态,CCK-8试剂盒检测细胞增殖情况,MDC荧光染色观察细胞自噬水平,Western-Blot检测自噬蛋白LC3的表达,qRT-PCR检测自噬基因Beclin-1的表达。结果:经丹参多酚酸盐处理后,胶质瘤U251细胞形态出现异常,CCK-8法检测发现细胞增殖能力下降,MDC荧光染色观察可见U251细胞质内有大量自噬体和自噬溶酶体,其自噬水平随丹参多酚酸盐作用时间的延长逐渐增强。Western-Blot法检测发现自噬相关蛋白LC3的表达随着丹参多酚酸盐浓度增加而增加,且随着药物浓度的增加,LC3-Ⅰ向LC3-Ⅱ转化增多。qRT-PCR法检测发现自噬相关基因Beclin-1 mRNA的表达随着丹参多酚酸盐浓度增加而增加。结论:丹参多酚酸盐可诱导胶质瘤U251细胞发生自噬,其诱导U251细胞发生自噬的机制可能与上调Beclin-1的表达有关。Objective： To observe autophagy of glioma U251 cells induced by depsides salts from Salvia Miltiorrhiza and explore the possible mechanism.Methods： The glioma U251 cell treated with depsides salts from Salvia Miltiorrhiza was used in this study.Cell morphology was dynamicly observed by inverted microscope.Cell viability was determined by cell counting kit-8.The intensity of autophagy was analysed by monodansylcadaverine（ MDC） fluorescent staining.The level of LC3 protein was measured by Western-Blot.The level of Beclin-1 mRNA was measured by qRT-PCR.Results： After incubation with different concentrations of depsides salts from Salvia Miltiorrhiza,the morphology of U251 cell was abnormal and the viability of U251 cell were obviously inhibited by cell counting kit-8.Autophagosomes and autolysosomes were observed in the depsides salts from Salvia Miltiorrhiza treated glioma U251 cell by MDC staining,intensity of autophagy was in a time-dependent manner.The expression levels of related protein LC3 and gene Beclin-1 were correlated with concentrations of depsides salts from Salvia Miltiorrhiza,and LC3-I to LC3-Ⅱincreased.Conclusion： Depsides salts from Salvia Miltiorrhiza may induce autophagy in glioma U251 cell,the mechanism may be related to raise the expression of Beclin-1.

关 键 词：丹参多酚酸盐 胶质瘤 自噬 LC3 BECLIN-1 

分 类 号：R284.1[医药卫生—中药学]