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目的研究土槿皮乙酸(pseudolaric acid B,PAB)对大鼠脑胶质瘤C6细胞增殖的抑制作用。方法采用MTT法检测0、0.1、0.2、0.4、0.8、1.6、8、16μg/ml PAB对C6细胞增殖能力的影响;流式细胞术检测碘化丙啶(PI)染色后细胞周期的变化;皮下注射C6细胞建立C6种植瘤大鼠模型并测量种植瘤体积;免疫组化法测定大鼠脑胶质瘤C6细胞种植瘤组织中增殖细胞核抗原(PCNA)的表达变化。结果体外实验结果表明,PAB呈剂量依赖性地抑制胶质瘤C6细胞生长,半数致死剂量(IC50)为0.03μg/ml,同时,可使C6细胞周期阻滞于G2/M期;体内实验结果表明,PAB给药后大鼠种植瘤体积有所降低,种植瘤组织中PCNA表达呈下降趋势。结论 PAB可抑制大鼠脑胶质瘤C6细胞增殖,阻滞细胞周期中G2/M期、下调PCNA表达为其可能的作用机制。
Objective To study the inhibitory effect of pseudolaric acid B (PAB) on the proliferation of glioma C6 cells in rats. Methods MTT assay was used to detect the effect of PAB on the proliferation of C6 cells at 0, 0.1, 0.2, 0.4, 0.8, 1.6, 8 and 16 μg / ml. The cell cycle was detected by flow cytometry after propidium iodide (PI) staining. C6 rat model was established by subcutaneous injection of C6 cells and the volume of implanted tumor was measured. The expression of proliferating cell nuclear antigen (PCNA) in C6 glioma implanted tumor tissue was detected by immunohistochemistry. Results The results of in vitro experiments showed that PAB inhibited the growth of C6 glioma cells in a dose-dependent manner with an IC50 of 0.03 μg / ml. In addition, the cell cycle arrest of C6 cells in G2 / M phase was inhibited by PAB. The results showed that the volume of implanted tumor decreased after PAB administration, and the expression of PCNA in implanted tumor showed a decreasing trend. Conclusion PAB can inhibit the proliferation of C6 glioma cells, arrest the G2 / M phase in the cell cycle, and down-regulate the expression of PCNA as its possible mechanism.