摘要
目的:研究喜树碱(CPT)对刀豆蛋白A(ConA)介导的小鼠T淋巴细胞体外活化、增殖及细胞周期的影响。方法:以ConA刺激小鼠T淋巴细胞,建立小鼠T淋巴细胞活化、增殖的模型,以不同浓度的CPT作用于该模型,流式细胞术检测T细胞早期活化标志CD69分子的表达;以活体染料羧基荧光素乙酰乙酸(CFDA-SE)染色流式细胞术分析CPT在ConA刺激下小鼠淋巴细胞的增殖相关指数(PI);以碘化丙啶染色流式细胞术分析细胞周期的分布情况。结果:ConA作用6h后流式细胞术分析显示CD69的表达率为(58.88±0.55)%,不同浓度的CPT组(10nmol/L、20nmol/L、50nmol/L、100nmol/L)CD69的表达率分别为(55.48±0.98)%、(54.67±1.05)%、(50.40±0.82)%、(42.47±1.32)%,均可明显抑制CD69的表达(P<0.01);ConA刺激48h后流式细胞术分析显示,不同浓度的CPT组(10nmol/L、20nmol/L、50nmol/L、100nmol/L)的增殖指数(PI)明显低于对照组(P<0.05);细胞周期分析显示刺激48h后不同浓度的CPT组(10nmol/L、20nmol/L、50nmol/L、100nmol/L)均出现明显的凋亡峰(apoptosispeak,AP),sub-G1期、G0/G1期细胞的比率明显高于ConA刺激组(P<0.01)。结论:CPT可明显抑制ConA刺激的T淋巴细胞的活化、增殖,同时使淋巴细胞阻滞于G0/G1期。
AIM： To study the effects of camptothecin （CPT） on the activation, proliferation and cell - cycle distribution of the mouse T lymphocytes stimulated by concanvalin A （ConA） in vitro. METHODS： A model of T cell activation and proliferation was established by stimulated the cells with Con A. T cells were treated with different concentrations of CPT. The expression of CD69, the early marker of CD3^＋ T cell activation, was measured by FACS. The proliferation index was determined by carboxyl fluorescin diacetate succinmidyl ester by flow cytometry. The cell - cycle distribution was analyzed by propidium iodide staining. RESULTS： After stimulation with Con A for 6 h, the activation rate of CD69^＋ T cell in Con A group was （58. 88±0. 55 ） %. The percentages of CD69 positive cells were （55.48±0. 98 ） %, （54. 67±1.05） %, （50. 40±0. 82） %, （42.47±1.32） %, correspond to the treatments with different concentrations of CPT （ 10 nmol/L, 20 nmol/L, 50 nmol/L, 100 nmol/L）, respectively. After 48 h treatment with Con A, the proliferation index in different concentrations of CPT treatment （ 10 nmol/L, 20 nmol/L, 50 nmol/L and 100 nmol/L） exerted a definite inhibitory effect on the proliferation （ P 〈0. 01 ）. Moreover, the cell-cycle distribution analysis showed that apoptosis peak was observed in different concentrations of CPT treatment after 48 h cultured with Con A. CONCLUSION： CPT significantly inhibits the early stages of the Con A-induced T cell activation and proliferation, and detents the T lymphocytes in G0/G1 phase.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2008年第6期1178-1182,共5页
Chinese Journal of Pathophysiology
基金
国家自然科学重点基金资助项目(No30230350)
广东省自然科学基金资助项目(No5300413)
