目的 探讨红霉素对弹性蛋白肽暴露下CD4^+T细胞向Th17细胞分化的影响。方法 从小鼠脾脏经免疫磁珠分选出CD4^+T细胞, 随机分为空白对照组、弹性蛋白肽组和弹性蛋白肽加红霉素干预组。弹性蛋白肽组和弹性蛋白肽联合红霉素处理组加入终浓度30 μg/mL的弹性蛋白肽,弹性蛋白肽联合红霉素处理组在弹性蛋白肽处理的基础上,另外加入100 μg/mL的红霉素。3组细胞在无血清培养液培养24 h, 用流式细胞术检测Th17的百分比, 荧光定量PCR检测维甲酸相关孤儿核受体γt (RORγt)mRNA表达, Western blot法检测核因子κB(p65)[NF-κB(p65)]及信号转导子和转录激活子3(STAT3)蛋白相对含量。结果 弹性蛋白肽组CD4^+IL-17^+细胞的百分比为(11.32±2.34)%、对照组为(5.21±1.36)%; qRT-PCR发现RORγt mRNA表达比对照组明显增加, Western blot法检测发现NF-κB(p65)和STAT3蛋白表达比对照组增强。与弹性蛋白肽组比较, 弹性蛋白肽联合红霉素处理组的CD4^+IL-17^+细胞的百分比、RORγt mRNA表达、NF-κB(p65)和STAT3蛋白表达明显减少。结论 红霉素可抑制弹性蛋白肽诱导CD4^+T细胞向Th17分化。
Objective To investigate the effect of erythromycin on differentiation into T helper 17 (Th17) cells from CD4^ + T cells exposed to elastin peptides. Methods The CD4^+ T cells from spleen of male BALB/c mice by magnetic bead sorting were randomly divided into the control group (group A), the elastin peptides group (group B) and erythromycin group(group C). The cells in group B and C were cultured in serum-free medium supplemented with 30 μg/mL elastin peptides. The cells in group C were additionally administrated with erythromycin at a dose of 100 μg/mL. All of the CD4^+T cells were cultured in serum-free culture solution for 24 hours. The number of the Th17 cells was detected by flow cytometry and the expression of retinoic acid related orphan nuclear receptor γt (RORγt) mRNA was measured by fluorescence quantitative PCR in each group. The expressions of NF-κB ( 1065 ) and signal transducer and activator of transcription 3 (STAT3 ) were analyzed by Western blotting. Results The Th17 cells in group B [ ( 11.32 ±2.34) % ] increased as compared with that in group A [ (5.21 ± 1. 36)% ], and the expression of RORγt mRNA in group B was higher than that in group A, Furthermore, the expressions of NF-κB (1065) and STAT3 proteins in group B increased significantly as compared with those in group A. However, compared with group B, group C presented with the significantly decreased expressions of Th17 cells, RORγt mRNA, NF-κB(p65) and STAT3 proteins. Conclusion Erythromycin can suppress the differentiation into Th17 cells from CD4^+T cells exposed to elastin peptides.