摘要
目的 :分析小檗碱 (Ber)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的机制。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 ,以流式细胞仪分析小檗碱对淋巴细胞增殖率的影响 ;以WST 1检测淋巴细胞的增殖情况 ;用碘化丙锭 (PI)染色分析细胞周期分布 ;细胞凋亡以Annexin V染色进行分析。结果 :CFSE染色分析显示 ,当浓度为 10 0、30和 10 μmol L时 ,Ber无论对ConA或PDB +Ion诱导的小鼠淋巴细胞增殖 ,都具有明显的抑制作用 (P <0 0 5 )。以WST 1分析细胞增殖情况 ,也获得相似的结果。对细胞周期分析表明 ,随着小檗碱浓度的增加 ,处于亚二倍体峰的细胞数增加 ,处于S和G2 M期的细胞数则减少 ,而G0 G1 期的细胞数基本不变。Annexin V染色法结果表明 ,30 μmol LBer组Annexin V单阳性区域的百分率为 ( 7 13± 1 0 8) % ,与PDB +Ion阴性组的百分率 ( 2 4 9± 0 2 5 ) %比较有显著的差异 (P <0 0 1)。结论 :小檗碱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。同时 ,Ber可以诱导体外培养的小鼠淋巴细胞发生凋亡。
Objective:To investigate the effect of berberine (Ber) on the proliferation and the cell-cycle distribution of the mouse lymphocytes,in order to elucidate the molecular and cellular mechanism for the immunosuppressive effect of berberine.Methods:Lymphocytes were isolated from lymphoid nodes of C57BL/6 mouse.After the staining with CFSE,mouse lymphocytes was stimulated with polyclonal activators,ConA or PDB plus Ion. The proliferation rate of mouse lymphocytes was analyzed with flow cytometry. WST-1 test was adopted to analyze the proliferation of mouse lymphocytes. The distribution of the cell-cycle was analyzed by PI staining together with flow cytometry analysis. Annexin-V staining was used to detect the apoptotic cell death of lymphocytes.Results:CFSE staining showed that 100,30 and 10 μmol/L Ber could significantly inhibit the proliferation of mouse lymphocytes induced by ConA or by PDB plus Ion. And similar results were obtained with WST-1 assay. The inhibitory effect was dose-dependent.Flow cytometry analysis of the cell-cycle distribution revealed that the cell number in sub-G 0/G 1 peak (apoptotic peak) was increased,while the cell numbers in S and G 2/M was decreased as the concentration of Ber increased. However the cell number in G 0/G 1 phase had no significant changes under the concentration range.Conclusion:Ber can inhibit the proliferation of the mouse lymphocytes and block the progression of mouse lymphocytes into S and G 2/M phases.The inhibitory effect is cell-cycle specific.Moreover,Ber can induce mouse lymphocytes underlying apoptosis.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2004年第10期687-692,共6页
Chinese Journal of Immunology
基金
国家自然科学基金重点项目 (3 0 2 3 0 3 5 0 )
面上项目(3 0 3 7165 1)
国家重点基础研究发展规划项目(G19990 5 43 0 3和G2 0 0 0 5 70 0 6)资助
