南方医科大学学报

南方医科大学学报 ›› 2020, Vol. 40 ›› Issue (01): 118-124.doi: 10.12122/j.issn.1673-4254.2020.01.15

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白芍总苷抑制小鼠T淋巴细胞体外增殖促进活化诱导细胞死亡

熊传锋，齐杰莹，邓 蓉，谢丽芬，李长征，聂晓莉   

  • 出版日期:2020-02-19 发布日期:2020-01-20
  • 基金资助:

Total glucosides of paeony inhibits proliferation and promotes activation- induced cell death of mouse T cells in vitro

  

  • Online:2020-02-19 Published:2020-01-20

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摘要： 目的 研究白芍总苷（TGP）对小鼠T淋巴细胞体外增殖及活化诱导细胞死亡（AICD）的影响，并初探其免疫抑制机制。方 法 利用免疫磁珠分选技术，无菌分离小鼠脾脏T淋巴细胞，按照TGP溶液浓度（0、50、100、200 µg/mL）设为空白组、TGP低中高剂量组，anti-CD3/CD28刺激T淋巴细胞活化及增殖。流式细胞仪检测细胞早期活化标志、细胞活力及CFSE标记的T淋巴细胞增殖；RT-PCR检测Fas/FasL mRNA水平；流式细胞仪检测活化T淋巴细胞表面Fas/FasL蛋白表达量；Western blot检测细胞凋亡相关蛋白Bcl-2的表达。结果（1）与空白组相比，TGP中高剂量组在用药48 h后活细胞数目显著减少，活细胞比例明显下降（P<0.001）；（2）TGP组给药处理48 h后，T淋巴细胞分裂代数及已分裂比例显著降低，且抑制作用具有药物浓度依赖性（P<0.001）；（3）TGP处理组T淋巴细胞24 h后Fas mRNA表达量显著上调，并伴随着T淋巴细胞表面Fas蛋白的表达上调及胞内Bcl-2表达下调（P<0.01）。结论 TGP显著抑制T淋巴细胞增殖，并可通过上调Fas、下调Bcl-2的表达促进活化诱导的T淋巴 细胞死亡（AICD）。

Abstract: Objective To investigate the effects of total glucosides of paeony (TGP) on the proliferation and activation-induced cell death of mouse T cells and the mechanism underlying the immunosuppressive effects of TGP. Methods Purified total T cells isolated from the spleen of C57BL/6 mice were treated with TGP at 0, 50, 100, or 200 µg/mL and stimulated by anti-CD3/ CD28. Flow cytometry was performed to detect the cell death and the proliferation of CFSE-labeled T cells. The expression of Fas/FasL mRNA was detected using RT-PCR, and flow cytometry was used to analyze the expression of Fas/FasL proteins on activated T cells. Western blotting was used to detect the expression of Bcl-2 in the cells. Results TGP treatment for 48 h significantly reduced the total number and percentage of viable T cells and dose-dependently lowered the percentage of divided T cells. TGP treatment obviously up-regulated the cellular expression of Fas mRNA, enhanced Fas expression on the surface of the T cells, and decreased the expression level of Bcl-2 protein in the cells. Conclusion TGP significantly inhibits proliferation and promotes activation-induced cell death of mouse T cell by increasing the expression of Fas and downregulating the expression of Bcl-2.