端粒重复序列结合蛋白1与2在系统性红斑狼疮患者中的表达变化及其临床意义

Expression of telomeric-repeat binding factor-1 and telomeric-repeat binding factor-2 mRNA and protein in the peripheral blood mononuclear cells of patients with systemic lupus erythematosus

目的探讨端粒重复序列结合蛋白（TRF）1和TRF2在SLE发病中可能的作用。方法①收集107例SLE患者（根据病情是否活动分为活动期组40例与稳定期组67例，根据是否合并肾脏损害分为肾损组46例及无肾损组61例）、41名健康体检者为健康对照组的外周血、实验室资料及临床资料。②采用实时荧光定量多聚酶链反应（RT-qPCR）及蛋白质印迹法检测不同组别TRF1、TRF2转录水平和蛋白表达水平。③采用SPSS19．0软件行独立样本t检验、单因素方差分析及Kruskal．Wallis非参数检验，Speannan相关分析进行统计分析。结果①TRF1、TRF2转录及蛋白水平在SLE活动期组（TRF1：0．0031±0．0033；TRF2：0．0105±0．0648）、肾损组（TRF1：0．0023±0．0026；TRF2：0．0043±0．0033）分别高于稳定期组（TRF1：0．0012±0．0011；TRF2：0．0042±0．0086）、无肾损组（TRF1：0．0013±0．0018；TRF2：0．0034±0．0072）及健康对照组（TRF1：0．0012±0．0030；TRF2：0．0034±0．0027）比较差异有统计学意义（P均〈0．05），健康对照组与稳定期组、无肾损组分别比较差异无统计学意义；②Spearman相关性分析发现在SLE患者中TRF1转录水平与TRF2（r=0.356，P〈0.01）、ESR（r=0.365，P〈0.05）正相关，TRF2转录水平与TRF1（r=0．356，P〈0．01）、SLEDAI评分值（r=0．270，P〈0．05）、ESR（r=0.304，P〈0．05）、肌酐（r=0．258，P〈0.05）及尿蛋白定量（24h）（r=0.344，P〈0.05）正相关。结论TRF1、TRF2在SLE患者异常表达，提示其可能参与了SLE的发生发展；TRF2与SLEDAI评分、尿蛋白定量（24h）呈正相关，提示TRF2可能可作为SLE病情活动及肾脏榻害的生物标记。

Objective To investigate the mRNA and protein expression levels of telomeric-repeat binding factor-1 （TRF1） and TRF2 in the peripheral blood mononuclear cells （PBMCs） of patients with systemic lupus erythematosus （SLE）, and the relations between these gene expression levels and clinical data of SLE patients were explored. Methods According to disease activity, these SLE patients were divided into the active group （40 cases） and the stable group （67 cases）. These patients were also grouped as renal damage group （46 cases） and renal damage-free group （61 cases） based on their renal conditions. Healthy individuals （41 cases） were also included as control. Real-time quantitative polymerase chain reaction （RT-qPCR） was employed to study the mRNA expression of TRF1 and TRF2. The protein levels of TRF1 and TRF2 were measured by Western Blot （WB）. Independent-Samples t test or one-way analysis of variance （ANOVA） in conjunction with the Least-Significant Difference method （LSD method） wasperformed if the data were in normal distributions; otherwise, the Kruskal-Wallis test was applied. Spearman＇s correlation analysis was also used for statistical analysis. Results The mRNA and protein expression levels of TRF1 and TRF2 in the PBMCs of the active group （TRFh 0.003 1-＋0.003 3; TRF2：0.010 5±0.064 8） and renal damage group （TRFh 0.002 3 ±0.002 6; TRF2：0.004 3 ±0.003 3） were significantly increased compared to the stable group （TRFh 0.001 2±0.001 1; TRF2：0.004 2±0.008 6）, the renal damage-free group （TRFh 0.001 3±0.001 8; TRF2： 0.003±0.007 2） and healthy （TRFI： 0.001 2±0.003 0; TRF2：0.003 4±0.002 7） individuals respectively （P〈0.05）. In SLE patients, the expression levels of TRF1 mRNA were correlated with erythrocyte sedimentation rate （r=0.365, P〈0.05）; the expression levels of TRF2 mRNA were correlated with SLEDAI score （r=0.270, P〈0.05）, erythrocyte sedimentation rate （r=0.304, P〈0.05）, creatinine （r=0.258, P〈0.05） and 24-hour urinary protein （r=0.344, P〈0.05）. Conclusion Altered expression of TRF1 and TRF2 might be involved in the pathogenesis of Systemic lupus erythematosus. The positive correlation between TRF2 and SLEDAI score, 24-hour urinary protein suggest that TRF2 might be usedas a biomarker for disease activity or renal damage in SLE.