摘要
分别采用MTT法、CFSE染色结合流式细胞术、双色荧光抗体染色结合流式细胞术、PI染色法检测去铁胺(DFO)对小鼠T淋巴细胞的药物毒性、对T淋巴细胞增殖、对Con A刺激下的小鼠T细胞CD69表达和淋巴细胞周期的影响。研究结果表明:DFO对淋巴细胞的毒性T细胞体外增殖、活化和细胞周期无影响;不同浓度的DFO(5、10、20、40μmol/L)能显著抑制受Con A刺激48 h和72 h后的T淋巴细胞增殖;低浓度的DFO(5、10μmol/L)对小鼠T淋巴细胞CD69的表达没有影响,高浓度(20、40μmol/L)的DFO可抑制T淋巴细胞CD69的表达;Ph能将T淋巴细胞抑制在G0/G1期。说明DFO除可通过抑制T淋巴细胞活化外,可能还通过其他机制抑制T淋巴细胞增殖,且DFO可以将淋巴细胞周期抑制在G0/G1期,可望发展成为一种新的免疫抑制干预药物。
In this paper, the cytotoxiciy of DFO to lymphocytes was evaluated using MTT assay, carboxyfluorescein diacetatesuccinimidyl ester （CFDA-SE） staining plus flow cytometry assay was employed to investigate the effect of DFO on the proliferation of the lymphocytes, the expression levels of CD69 on T lymphocytes stimulated by Con A were evaluated with flow cytometry after staining with fluorescent monoclonal antibody, and Cell-cycle distribution of T lymphocytes was also analyzed by propidium iodide staining. The results showed that DFO induced-cytotoxicity had little impact on the reliability of our experiment data; DFO （5,10,20,40 μmol/L） significantly inhibited the proliferation of T lymphocytes stimulated by Con A for 48 h and 72 h;DFO at low concentrations （5,10 μmol/L ） exerted no influence on the expression of CD69, however, DFO at high concentration （20, 40 μmol/L ） could inhibit the expression of CD69. The cell cycle distribution analysis revealed that DFO could induce a cell cycle arrest in G0/G1 phase. We can draw conclusion that DFO may use other mechanism except the inhibition of activation of T lymphocyte, to inhibit the proliferation of lymphocyte, it can induce a cell cycle arrest in G0/G1 phase. This effect demonstrates that DFO has the potential to be developed into a new immuo-intervention drug.
出处
《广东农业科学》
CAS
CSCD
北大核心
2012年第16期165-168,共4页
Guangdong Agricultural Sciences
基金
国家"973"计划项目(2006CB504201
2004CB720100)
国家自然科学基金(30500466)
广东省自然科学基金(5300419)
广东省主体科研机构创新能力建设专项"华南农业基因研究与创新平台建设"(2011)
