雷公藤红素对Raji细胞表面超微结构与增殖活性的影响

Effect of tripterine on surface ultrastructure and proliferative activity of Raji cells

目的研究雷公藤红素对Burkitt淋巴瘤细胞株Raji的表面超微结构及其增殖活性的影响。方法选用不同浓度(0.5、1.0、1.5、2.0μg/ml)的雷公藤红素作用于Raji细胞,用CCK8法测定细胞增殖情况,用原子力显微镜观察细胞形貌和表面超微结构变化,用Hoechst 33258染色及流式细胞仪检测细胞的凋亡,用流式细胞仪检测细胞周期的改变。结果CCK8测定表明,经不同浓度的雷公藤红素作用24 h后,细胞存活率从(80.67±2.08)%下降至(38.53±2.25)%,48 h后,细胞存活率从(74.17±3.20)%下降至(33.22±1.64)%,雷公藤红素对Raji细胞存活率的影响呈时间和浓度依赖性。原子力显微镜扫描示:正常Raji细胞呈圆形,表面光滑,经浓度为2.0μg/ml的雷公藤红素处理24 h和48 h后,Raji细胞开始坍塌,超微结构显示细胞膜表面粗糙、凹凸不平。Hoechst 33258染色可见凋亡细胞;流式细胞术检测显示不同浓度的雷公藤红素作用Raji细胞24 h后,细胞凋亡率从(3.50±1.73)%增加到(38.27±6.05)%,雷公藤红素对Raji细胞凋亡率的影响呈浓度依赖性。流式细胞术对细胞周期的检测表明1.5μg/ml雷公藤红素作用Raji细胞24 h,可使S期细胞增加,与对照组比较差异有统计学意义(P<0.05)。结论雷公藤红素可改变Raji细胞膜超微结构,可通过诱导Raji细胞凋亡而抑制其增殖。

Objective To investigate the effect of tripterine on surface ultrastructure and proliferative activity of Burkitt lymphoma cell line Raji.Methods Raji cells were treated with different concentrations（0.5,1.0,1.5,and 2.0 μg/ml） of tripterine.Then the proliferation of Raji cells was determined by CCK8 assay,the cell membrane ultrastructure was analyzed by atomic force microscope,the apoptosis of cells was determined by Hoechst 33258 staining and flow cytometric analysis,and the cell cycle was assayed by flow cytometry.Results CCK8 assay showed that the survival rate of cells decreased from（80.67 ± 2.08） % to（38.53 ± 2.25） % 24 h after treatment with different concentrations of tripterine.The cell survival rate decreased from（74.17 ± 3.20） % to（33.22 ± 1.64） % 48 h after treatment.Tripterine significantly inhibited the proliferation of Raji cells and the inhibition was in a time-and concentration dependent manner.Atomic force microscope scan showed that the untreated Raji cells were round,with relatively smooth surface.After Raji cells were treated with 2.0 μg/ml tripterine for 24 h and 48 h,the ultrastructure of the cell membrane was collapsed,and the cell surface was rough and uneven.Hoechst 33258 staining demonstrated apoptotic cells.Apoptotic rate of the Raji cells increased from（3.50 ± 1.73） % to（38.27 ± 6.05） % 24 h after treatment with different concentrations of tripterine.Flow cytometry analysis showed that 24 h after treatment with 1.5 μg/ml tripterine,S phase Raji cells were significantly increased compared with control group（P〈0.05）.Conclusion Our results demonstrate that tripterine can alter the cell membrane ultrastructure of Raji cells and can inhibit Raji cell proliferation through inducing cell apoptosis.