[1]王旭,杨加彩,宋亚军,等.蛋白酶体抑制剂硼替佐米对同种反应性T细胞凋亡的影响[J].第三军医大学学报,2019,41(20):1926-1931.
 WANG Xu,YANG Jiacai,SONG Yajun,et al.Effects of Proteasome inhibitor bortezomib on apoptosis of murine alloreactive T cells in vitro [J].J Third Mil Med Univ,2019,41(20):1926-1931.
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蛋白酶体抑制剂硼替佐米对同种反应性T细胞凋亡的影响(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
41卷
期数:
2019年第20期
页码:
1926-1931
栏目:
基础医学
出版日期:
2019-10-30

文章信息/Info

Title:
Effects of Proteasome inhibitor bortezomib on apoptosis of murine alloreactive T cells in vitro
 
作者:
王旭杨加彩宋亚军杨其顺黄赤兵
陆军军医大学(第三军医大学)第二附属医院泌尿外科;陆军军医大学(第三军医大学)第一附属医院全军烧伤研究所,创伤、烧伤与复合伤国家重点实验室
Author(s):
WANG Xu YANG Jiacai SONG Yajun YANG Qishun HUANG Chibing

Department of Urology, Second Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400037; State Key Laboratory of Trauma, Burn and Combined Injury, Institute of Burn Research, First Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400038, China

关键词:
硼替佐米T细胞细胞增殖细胞周期细胞凋亡细胞因子
Keywords:
bortezmoib T cells proliferation cell cycle apoptosis cytokines
分类号:
R392.4;R392.5;979.5
文献标志码:
A
摘要:

目的 观察蛋白酶体抑制剂硼替佐米(bortezomib, BTZ)对同种反应性T细胞凋亡的影响。方法 建立单向混合淋巴细胞培养体系。细胞培养24 h后加不同浓度硼替佐米培养12 h,CCK-8检测细胞增殖;按细胞混合培养时间分组,每组设实验组和对照组,实验组加0.01 μmol/L 硼替佐米培养12 h,流式细胞术检测T细胞的凋亡、细胞周期和活化相关表面标志CD25、CD69,Western blot检测凋亡相关蛋白裂解半胱天冬酶3(Cleaved caspase 3)表达和周期蛋白D1(Cyclin D1)表达;ELISA检测IL-4、TNF-α和IFN-γ含量。结果 与对照组比,实验组T细胞增殖率降低;实验组凋亡T细胞占比增加、Cleaved caspase 3表达量升高,且与硼替佐米干预前细胞共培养时间呈正相关(P<0.05);G1期T细胞比例增加,周期蛋白D1表达量降低;CD25、CD69表达下调(P<0.05);实验组IL-4、TNF-α和IFN-γ浓度降低。结论 硼替佐米可抑制同种反应性T细胞增殖,诱导活化T细胞凋亡,并降低T细胞的细胞因子分泌。

Abstract:

ObjectiveTo observe the effects of the proteasome inhibitor bortezomib (BTZ)  on the apoptosis of murine alloreactive T cells  in vitro. MethodsIn a one-way mixed lymphocyte culture system, the T lymphocytes from C57BL/6 mice were co-cultured with the splenocytes of BALB/c mice for 24 h. After treatment of the cells with different concentrations of bortezomib for 12 h, the changes in the cell proliferation were assessed using CCK-8 assay. The responses of the co-cultured cells to different culture times and a 12 h treatment with 0.01 μmol/L bortezomib were examined by detecting the changes in the cell apoptosis, cell cycle and expression of the activation-associated markers CD25 and CD69 using flow cytometry; the expression of cleaved caspase-3 and cyclin D1 was detected using Western blotting, and the levels of interleukin-4 (IL-4), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in the supernatant were measured using enzyme-linked immunosorbent assay (ELISA). ResultsCompared with the control cells, the bortezomib-treated T cells showed an obviously decreased proliferative activity. Treatment with 0.01 μmol/L bortezomib significantly increased the percentage of apoptotic T cells and enhanced the expression of cleaved caspase-3 in positive correlation with the co-culture time prior to bortezomib treatment (P<0.05). The proportion of T cells at G1 phase increased and the expression of cyclin D1 decreased significantly in bortezomib-treated cells (P<0.05), which also showed down-regulated expressions of CD25 and CD69 and lowered levels of IL-4, TNF-α and IFN-γ in the supernatant (P<0.05). ConclusionBortezomib can inhibit the proliferation of murine alloreactive T cells in vitro, induce apoptosis and reduce cytokine production in the activated T cells.

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更新日期/Last Update: 2019-10-25