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目的 观察柴胡皂甙d(SSd)对HL 6 0细胞糖皮质激素受体 (GR)mRNA表达的调节作用以及对细胞凋亡的影响。方法 选用从柴胡中提取的单体成分SSd ,以3 H 胸腺嘧啶掺入法观察SSd对细胞生长的抑制作用 ,DNA凝胶电泳及流式细胞术分析细胞凋亡 ,用Northernblot分析GRmRNA的改变。结果 经SSd作用后 ,HL 6 0细胞3 H 胸腺嘧啶掺入率明显降低 ,呈时间和剂量依赖关系 ,10 μg/mlSSd处理 48小时作用最明显 ,6 0小时时出现典型的DNA片段梯形带 ,流式细胞仪分析细胞阻滞于G1期并出现凋亡峰 ;GRmRNA表达增加。结论 SSd可上调HL 6 0细胞GRmRNA表达并诱导细胞凋亡。
Objective To observe the effect of saikosaponin d (SSd) on glucocorticoid receptor (GR) mRNA expression in HL 60 cells and its effect on cell apoptosis. Methods SSd was extracted from Radix Bupleurum with monomeric components SSd. The inhibitory effect of SSd on cell growth was observed by 3 H thymidine incorporation method. Cell apoptosis was analyzed by DNA gel electrophoresis and flow cytometry. Changes of GR mRNA were analyzed by Northern blot analysis. . Results After treatment with SSd, the incorporation rate of 3 H thymidine in HL 60 cells was significantly decreased in a time- and dose-dependent manner. The effect of 10 μg/ml SSd treatment was most obvious at 48 hours, and a typical DNA ladder was observed at 60 hours. Flow cytometry analysis of cell arrest in the G1 phase and the appearance of apoptotic peak; GR mRNA expression increased. Conclusion SSd can upregulate GR mRNA expression and induce apoptosis in HL 60 cells.