摘要
目的探讨蛋白酶体抑制剂硼替佐米(bortezomib)对混合淋巴细胞培养体系中细胞凋亡、细胞因子水平的影响。方法体外建立单向混合淋巴细胞培养(MLC)体系,分别用2、4、8 nmol/L的硼替佐米干预反应体系,在不同的时间点收集细胞,流式细胞仪检测细胞凋亡率、ELISA法检测培养上清液中白细胞介素2(IL-2)、γ干扰素(IFN-γ)、肿瘤坏死因子α(TNF-α)的含量。结果硼替佐米作用于细胞12、24、36 h后细胞凋亡率逐渐增加,8 nmol/L硼替佐米作用36 h细胞凋亡率为(61.67±3.21)%;硼替佐米作用24 h后上清液中IL-2、IFN-γ、TNF-α浓度减小,8 nmol/L组的IL-2、IFN-γ、TNF-α浓度分别为(88.27±2.76)ng/L、(57.36±2.08)ng/L、(22.19±0.88)ng/L。结论硼替佐米能诱导细胞凋亡,使混合淋巴细胞培养体系细胞分泌的Th1型细胞因子减少。
Objective To investigate the effect of bortezomib on the apoptosis and the cytokine levels of the mixed lymphocyte culture system. Methods One -way mixed lymphocyte culture (MLC) system was established in vitro and bortezomib was employed at the concentrations of 2, 4 and 8 nmol/L, respectively, to intervene MLC; the cells were harvested at different time points and flow cytometry was used to detect apoptosis rate. The supernatants were harvested and analyzed for IL - 2, IFN - γ, TNF - α secretion using a commercially available enzyme - linked immunosorbent assay (ELISA) kits. Results Bortezomib could effectively induce apoptosis of MLC system. 12, 24 and 36 hours after treatment with bortezomib, the apoptosis rate increased gradually. 36 hours after treatment with 8 nmol/L bortezomib, the apoptosis rate was 61.67 + 3.21%, and the level of IL- 2, IFN -γ, TNF -α in culture supernatants decreased; 24 hours after treatment with 8 nmol/L bortezomib, the level of IL - 2, IFN - β and TNF - α were 88.27 ± 2. 76 ng/L, 57.36 ± 2. 08 ng/L, 22.19 ±0.88 ng/L, respectively. Conclusion Bortezomib could induce apoptosis of MLC system, with a decrease in the production of ThI cytokines.
出处
《徐州医学院学报》
CAS
2008年第10期655-658,共4页
Acta Academiae Medicinae Xuzhou
基金
江苏省卫生厅科研项目(H200722)