硫酸酯化天麻多糖抗脑胶质瘤活性的研究

Studies on Anti-C6 Glioma Cells Activity of Sulfated Polysaccharides from Gastrodia Elata Blume

目的:研究硫酸酯化天麻多糖(sulfated gastrodia elata polysaccharides,GEPs)抗脑胶质瘤的活性。方法:将大鼠脑胶质瘤细胞(C6)实验分为空白对照组和GEPs各剂量组(0.10,0.25,0.50,1.00,1.50 mg·ml^-1),采用CCK-8法检测细胞存活率,并观察细胞形态变化;采用DAPI荧光染色法观察细胞凋亡形态;采用划痕实验检测不同浓度GEPs对细胞迁移能力的影响。结果:GEPs各剂量组作用于C6胶质瘤细胞24 h和48 h时可显著降低细胞生存率(P<0.05),并呈浓度依赖性(P<0.01)。倒置显微镜下可见,随着GEPs浓度的上升,C6细胞体积变小,细胞间隙变大,失去原有的细胞形态,并且贴壁不良,产生大量漂浮物。DAPI染色观察到C6细胞数量明显锐减,细胞核缩小,凋亡的细胞明显增多。细胞划痕实验表明,GEPs各剂量组作用于C6细胞24 h后,愈合率逐渐下降,并呈浓度依赖性(P<0.05)。结论:硫酸酯化天麻多糖能显著抑制神经胶质瘤细胞生长的活性,并可有效抑制神经胶质瘤细胞的迁移。

Objective:To explore the anti-C6 glioma cells activity of sulfated polysaccharides from Gastrodia Elata Blume(GEPs).Methods:The C6 glioma cells were divided into the control group and GEPs groups(0.10,0.25,0.50,1.00 or 1.50 mg·ml^-1).The cell growth inhibition in vitro was assessed by CCK-8 assay.The morphological changes of C6 cells pre-and posttreatment were observed under a microscope.The cell apoptosis was observed through DAPI fluorescent staining under a microscope.The effects of GEPs at different concentrations on the cell migration were detected by scratch test.Results:With the pretreatment of GEPS,the survival rate of C6 cells decreased significantly(P<0.05)in a dose-dependent manner(P<0.05).Under an inverted microscope,C6 cells incubated with GEPS were smaller,and the cell gap became larger and the original cell morphology was lost,and the adhesion was loose with lots of cells floating.DAPI staining showed that the number of C6 cells decreased significantly,the nucleus shrank,and the number of apoptotic cells increased significantly.The scratch test showed that the healing rate of C6 cells decreased gradually at 24 h in a dose-dependent manner(P<0.05).Conclusion:GEPs can effectively restrain the proliferation activity of C6 cells and restrain C6 cells migration.