三七提取物对小鼠淋巴细胞体外增殖和细胞周期的影响

Effects of Sanchi extract on the in vitro proliferation and cell-cycle distribution of mouse lymphocytes

目的分析三七提取物(SE)对小鼠淋巴细胞体外增殖和细胞周期的影响,探讨其免疫抑制作用机制。方法分离小鼠淋巴结细胞,以CFSE染色,加入多克隆刺激剂刀豆蛋白(ConA)或者佛波醇酯(PDB)和离子霉素(Ion)进行刺激,以流式细胞仪分析三七提取物对淋巴细胞增殖的影响;用碘化丙锭(PI)染色分析细胞周期分布。结果CFSE染色分析显示,SE对ConA或者佛波醇酯(PDB)和离子霉素(Ion)诱导的小鼠淋巴细胞增殖,具有明显的抑制作用(P<0.05),且呈剂量依赖性。对细胞周期分析表明,随着三七提取物浓度的增加,均能明显阻止淋巴细胞进入S期和G2/M期,而处于亚二倍体峰的细胞数基本不变。结论三七提取物对小鼠淋巴细胞的增殖有明显抑制作用,它能抑制淋巴细胞进入细胞分裂周期,这种抑制作用表现出明显的周期特异性。

Objective To investigate the effects of Sanchi extract （SE） on proliferation and cell-cycle distribution of mouse lymphocytes, in order to elucidate the molecular and cellular mechanism of the immunesuppressive effect of SE. Methods Lymphocytes were isolated from lymphoid nodes of Balb/c mouse. After the staining with CFSE, the mouse lymphocytes were stimulated with polyclonal activators Con A or PDB plus Ion. The proliferation index of mouse lymphocytes was analyzed with flow cytometry. The distribution of the cell-cycle was analyzed by PI staining together with flow cytometry analysis. Results In a dose-dependent manner, SE could significantly inhibit the proliferation of mouse lymphocytes induced by Con A or by PDB plus Ion. Flow cytometry analysis of the cell-cycle distribution revealed that the cell number in GO / G1 was increased, while the cell number in S and G2/M was decreased as the concentration of SE increased. However, the cell number in sub- GO/G1 peak （apoptotic peak） phase had no significant changes under the concentration range. Conclusion SE can inhibit the proliferation of the mouse lymphocytes and block the progression of mouse lymphocytes growing into S and G2/M phases. The inhibitory effect is cell-cycle specific.