论文部分内容阅读
目的:研究根皮素(Ph)对小鼠T淋巴细胞中期活化、细胞周期、Ca2+离子内流以及凋亡的影响,探讨将其发展为免疫干预药物的可能性。方法:双色荧光抗体染色结合流式细胞术(FCM)分析Ph对多克隆刺激剂(ConA)刺激下的小鼠T淋巴细胞CD25表达的影响;PI染色法检测Ph对T淋巴细胞周期的影响;Fluo-4/AM染色结合FCM分析Ph对T淋巴细胞Ca2+离子内流的影响;calcein/Co2+技术检测Ph对T淋巴细胞线粒体通透性转换(MPT)的影响,并用DIOC6(3)染色检测其对线粒体膜电势(MMP)的影响。结果:Ph(40,60,80μmol/L)对ConA刺激的T淋巴细胞CD25表达有显著的抑制作用(P<0.01);Ph能抑制T淋巴细胞Ca2+离子内流(P<0.01);细胞周期分析显示,Ph能将T淋巴细胞抑制在S期;且Ph能够诱导和加速MPT,并导致MMP的显著下降。结论:Ph对小鼠T淋巴细胞活化和Ca2+内流具有显著的抑制作用,并能将细胞抑制在S期,能通过线粒体途径诱导T淋巴细胞凋亡,可望发展成为一种新的免疫抑制药物。
Objective: To investigate the effects of Ph Ph on activation, cell cycle, Ca 2+ influx and apoptosis in T lymphocytes of mice and to explore the possibility of developing Phosphointerone into immune intervention drugs. Methods: The effect of Ph on CD25 expression of mouse T lymphocytes stimulated by polyclonal stimulator (ConA) was analyzed by dual color fluorescent antibody staining combined with flow cytometry (FCM). The effect of Ph on the T lymphocyte cycle was detected by PI staining. Fluo-4 / AM staining and FCM were used to analyze the effect of Ph on the influx of Ca2 + in T lymphocytes. The effect of Ph on mitochondrial permeability transition (TPR) was detected by calcein / Co2 + assay and DIOC6 (3) On mitochondrial membrane potential (MMP). Results: Ph (40, 60 and 80μmol / L) significantly inhibited the CD25 expression of ConA-stimulated T lymphocytes (P <0.01); Ph inhibited the influx of Ca2 + ions of T lymphocytes (P <0.01) Analysis shows that Ph can inhibit T lymphocytes in S phase; and Ph can induce and accelerate MPT, and lead to a significant decrease in MMP. CONCLUSION: Ph can significantly inhibit the activation of T lymphocytes and influx of Ca2 + in mice, and can inhibit the cells in S phase and induce the apoptosis of T lymphocytes through mitochondria pathway. It is expected to develop into a new immunosuppressive drug.