强直性脊柱炎患者外周血Th17、Treg、Th1免疫失衡及其与疾病活动相关性研究

Expression of transcription factor Th17,Treg and Th1 in peripheral blood from patients with ankylosing spondylitis and its correlation with disease activity

目的:初步探讨强直性脊柱炎(AS)患者外周血中Th17细胞、调节性T细胞(Treg)、Th1细胞免疫失衡及其与疾病活动的相关性。方法:分离AS患者(61例)和健康对照组(36例)外周血单个核细胞(PBMCS),应用Rt-PCR检测Th17、Treg、Th1的特异性转录因子RORC、FoxP3、T-bet的mRNA表达水平;流式细胞术检测CD4+CD25+FoxP3+T细胞、CD4+IL-17+Th细胞及CD4+Th1细胞的比率;ELISA分析外周血IL-17、IL-23、TNF-α、IL-6等蛋白水平。结果:Foxp3 mRNA表达量AS组显著低于健康对照组(0.66±0.37 vs 1.5±0.82,P<0.01)且与AS活动度指数(ASDAS)及IL-23蛋白水平互为强负相关(r=-0.46,P<0.01,r=-0.46,P<0.05);而RORC mRNA表达水平AS组显著高于健康对照组(1.33±0.77 vs 0.83±0.52,P<0.05),与ASDAS呈显著正相关(r=0.63,P<0.05)且与IL-17、IL-23、TNF-α蛋白水平呈强正相关(r=0.52,P<0.05;r=0.57,P<0.05;r=0.52,P<0.05);T-bet mRNA表达量AS组显著低于健康对照组(0.68±0.65 vs 2.21±1.29,P<0.01)且与RORC mRNA表达量显著负相关(r=-0.37,P<0.05);CD4+IL-17+Th17细胞比率AS组明显高于健康对照组(1.987±0.58 vs 0.647±0.33,P<0.05),CD4+CD25+FoxP3+T细胞及CD4+Th1细胞的比率AS组明显低于对照组(0.94±0.38 vs2.13±0.75;3.98±0.72 vs 6.85±1.36,P<0.05);IL-17、IL-23、TNF-α表达量AS组显著高于健康对照组(37.81±5.81 vs 8.68±0.88,540.89±46.95 vs 181.85±32.91,37.84±4.34 vs 7.48±2.04,P<0.01)且之间互为强正相关(r=0.88,0.92,0.95)。结论:AS患者存在Th17/Treg/Th1细胞平衡失调,且与AS病情活动密切相关,可能为AS的预防和治疗提供新思路。

Objective：To investigate the level of cell-specific transcription factor Foxp3,RORC and T-bet in peripheral blood from patients with ankylosing spondylitis（AS）and its correlation with disease activity.Methods： Separate AS patients＇ peripheral blood from the control group.The mRNA expression of FoxP3,RORC and T-bet were measured by real-time quantitative PCR（Rt-PCR）.The content of IL-17,IL-23,TNF-α and IL-6 were detected by ELISA.Results： The expression of Foxp3 mRNA in AS group was significantly lower than that of the control group（0.66 ± 0.37 vs 1.5 ± 0.82,P〈0.01）,and the Foxp3 mRNA level was highly negatively correlated with ASDAS and IL-23 level（r=-0.46,P〈0.01,r=-0.46,P〈0.05）;The expression of RORC mRNA in AS group was higher than that of the control group（1.33±0.77 vs 0.83±0.52,P〈0.05）,closely paralleling with the expression of IL-17,IL-23 and TNF-α（r=0.52,P〈0.05;r=0.57,P〈0.05;r=0.52,P〈0.05）,the expression of T-bet mRNA in AS group was significantly lower than that of the control group（0.68±0.65 vs 2.21±1.29,P〈0.01） and the RORC mRNA level was significantly negative correlated（r=-0.37,P〈0.05）.The ratio of CD4＋Th17 T cell in AS group was significantly higher than that of the control group（1.987±0.58 vs 0.647±0.33,P〈0.05）.The ratio of CD4＋CD25＋FoxP3＋T cell and the ratio of CD4＋Th1 cell of AS group were significantly lower than that of the control group（0.94±0.38 vs 2.13±0.75;3.98±0.72 vs 6.85±1.36,P〈0.05）.The expression of IL-17,IL-23 and TNF-α in AS group were significantly higher than that of the control group（37.81±5.81 vs 8.68±0.88,540.89±46.95 vs 181.85±32.91,37.84±4.34 vs 7.48±2.04,P〈0.01）,and the correlation were significantly positive（r=0.88,0.92,0.95）.Conclusion： The imbalance of the Treg/Th17 and Th1/Th17 cell exist in AS patients and they are significantly related to AS illness,which may provide news ways and new thinking to prevent and against AS.