摘要
目的探讨1,25(OH)_2D_3对Th17细胞分化的体外抑制作用。方法通过对分离出的脾淋巴细胞进行CD4+T细胞分选,将不同浓度的1,25(OH)_2D_3加入到CD4+T细胞中并在诱导因子的作用下诱导其分化。通过ELISA检测培养上清中IL-17A和IL-22的浓度;流式细胞仪分析Th17细胞的比例及RT-PCR检测Th17细胞分化的转录因子的表达。结果培养上清ELISA检测显示:与对照组相比,1,25(OH)_2D_3组的IL-17A和IL-22浓度随着加入1,25(OH)_2D_3浓度的增大而呈现逐渐降低的现象,差异且具有统计学意义(P<0.01);流式结果显示:诱导分化后对照组Th17细胞比例最多,1,25(OH)_2D_3组处理后Th17细胞比例降低,且呈剂量依赖性;RTPCR检测显示:与对照组相比,1,25(OH)_2D_3组IL-17A、IL-22,RORγt和RORα表达量随着剂量的增加而逐渐降低,并具有统计学意义(P<0.01)。结论体外实验表明,1,25(OH)_2D_3具有抑制Th17细胞分化的作用,表现为Th17细胞比例、分泌的细胞因子及特异的转录因子表达均减少。
objective To study and confirm the inhibition effect of 1,25（OH）_2D_3 on Th17 cells differentiation in vitro. Methods Lymphocytes were separated from spleen. Different concentrations of 1,25（OH）_2D_3 were added to the CD4＋T cells. Th17 cell differentiation was induced by CD4＋T Cell Isolation Kit. The level of IL-17 A and IL-22 were detected by ELISA. Flow cytometry was used to analyze the proportion of Th17 cells while RT-PCR was used to detect the expression of transcription factors of th17 cells. Results Cytokines detection in culture supernatant by ELISA showed that IL-17 A and IL-22 concentrations gradually decreased, with an increasing addition of 1,25（OH）_2D_3 concentration. The proportion of Th17 cells reduced in 1,25（OH）_2D_3 treatment group and presented a dose dependent. The m RNA levels of IL-17 A, IL-22, RORγt and RORα were significantly decrease in 1,25（OH）_2D_3 treatment group than that in control group（P〈0.01）. Conclusion 1,25（OH）_2D_3 plays a role in inhibiting the differentiation of Th17 cells, reduces the cytokines secretion（IL-17 A and IL-22） and expression of specific transcription factors（IL-17 A, IL-22, RORγt, RORα）.
出处
《肝胆胰外科杂志》
CAS
2016年第3期213-216,共4页
Journal of Hepatopancreatobiliary Surgery
基金
上海市卫生和计划生育委员会科研课题(20154Y0207)
