摘要
目的:探讨细胞外信号调节激酶1/2(ERK1/2)信号通路在小鼠T淋巴细胞增殖、周期和活化中的作用。方法:分离小鼠淋巴结细胞,藉多克隆刺激剂刀豆蛋白(ConA)或佛波醇酯(PDB)加离子霉素(Ion)刺激,流式细胞术分析ERK1/2信号通路的特异性阻断剂PD98059对小鼠T淋巴细胞增殖、周期和活化的影响。结果:活体染料羧基荧光素乙酰乙酸染色分析显示,不同浓度(5、10、20、30、40μmol/L)的PD98059对ConA诱导的T淋巴细胞增殖具有明显的抑制作用,呈现剂量依赖关系(r=0.985,P<0.01)。碘化丙锭染色分析表明,PD98059可阻止ConA刺激的T淋巴细胞进入S期和G2/M期,PD98059对PDB+Ion刺激的T淋巴细胞细胞周期的影响与ConA刺激相似,不同的是S期和G2/M期的变化较ConA作用更显著。荧光标记单克隆抗体染色显示,不同浓度的PD98059仅能轻微影响T淋巴细胞表面活化标志CD69和CD25的表达。结论:PD98059对小鼠T淋巴细胞的增殖有明显抑制作用,并阻止其进入S期和G2/M期,但不能明显抑制小鼠T淋巴细胞的早期和中期活化。
Aim： To investigate extracellular signal regulated kinase 1/2 （ERK1/2） signal pathway on proliferation, cell cycle and activation of mouse lymphocytes. Methods： Lymphocytes were isolated from lymph nodes in BALB/c mice and then stimulated with polyclonal activators, concanavalin （ConA） and phorboll 2,13-dibutyrate （PDB） plus ionomycin （Ion） . The proliferation, cell cycle and activation of T lymphocytes were analyzed using flow cytometry（FCM）. Results： Carboxy fluoresce in diacetate succin- imidyl ester （CFDA-SE） staining showed that 5, 10, 20, 30 and 40 p^mol/L PD98059 could significantly inhibit the proliferation of mouse T lymphocytes induced by ConA. The inhibitory effect appeared dosedependent（r =0. 985 ,P 〈0. 01 ）. The analysis of the cell cycle induced by ConA revealed that the cells were blocked to enter S and G2/M phases with the concentration of PD98059 increased. PD98059 had the similar effect on the cells treated with PDB plus Ion, but the changes of S phase and G2/M phase were more distinct. The fluorescence-conjugated mAb staining showed that PD98059 had only slight inhibition of the expression of CD69 on the surface of T lymphocytes and had no effect on the expression of CD25 on the surface of T lymphocytes. Conclusion： PD98059 can inhibit the proliferation of the mouse T lymphocytes and block them to enter S and G2/M phases, but it can not remarkably inhibit the activation of mouse T lymphocytes induced by ConA.
出处
《暨南大学学报（自然科学与医学版）》
CAS
CSCD
北大核心
2007年第4期345-350,共6页
Journal of Jinan University(Natural Science & Medicine Edition)
基金
国家自然科学基金资助项目(30471635)
广东省自然科学基金资助项目(04010451
5006033)
暨南大学引进优秀人才科研启动基金(51204058)
