TRAIL在系统性红斑狼疮发病机制中的作用及其相关性研究

The role and research of TRAIL in the pathogenesis systemic lupus erythematosus

目的探索肿瘤坏死因子相关凋亡诱导配体(TRAIL)在系统性红斑狼疮(SLE)细胞凋亡、自身抗体产生和疾病活动中作用。方法60例SLE患者及20名健康对照,反转录聚合酶链反应(RT-PCR)半定量分析外周血单个核细胞(PBMC)的TRAIL mRNA表达;比色法测定PBMC内Caspase-3活性表达;酶联免疫法(ELISA)测定PBMC内核小体水平、血清中可溶性TRAIL(sTRAIL)和抗核小体抗体(AnuA)浓度。结果狼疮活动组与稳定组及健康对照组相比,TRAILmRNA平均表达水平(0.82±0.07)vs(0.77±0.06)vs(0.75±0.05)、Caspase-3活性(0.53±0.27)vs(0.39±0.23)vs(0.35±0.18)、核小体浓度(U/ml)((3.09)vs(0.53)vs(0.77)及血清sTRAIL水平(μg/L)(0.88±0.74)vs(0.58±0.32)vs(0.48±0.28),差异均有显著性(P<0.05)。SLE患者TRAILmRNA平均表达水平与sTRAIL、Caspase-3活性及Caspase-3活性与核小体水平均呈显著正相关(P<0.05)。SLE患者TRAIL mRNA表达水平与SLE疾病活动指数(SLEDAI)积分数、抗dsDNA抗体、ESR、AnuA浓度呈正相关,与淋巴细胞计数及补体C4呈负相关;sTRAIL浓度与SLEDAI积分数呈正相关,与淋巴细胞计数呈负相关。核小体浓度与SLEDAI积分数、抗dsDNA抗体、AnuA浓度均呈显著正相关,与补体C3、C4水平呈负相关。结论SLE活动患者PBMC的TRAIL表达水平及血清sTRAIL水平增高,凋亡酶Caspase-3的活性增高,可能介导PBMC异常凋亡,使核小体释放增加,自身抗体水平增加,参与疾病活动。

Objective To investigate the effect of TRAIL on apoptosis,the production of autoantibodies,and disease activity in systemic lupus erythematosus （SLE）. Methods Sixty patients with SLE and 20 healthy controls were enlisted. Quantitative reverse transcription-polymerase chain reaction （RT-PCR） was used to measure the expression of TRAILmRNA on peripheral blood mononuclear ceils （PBMC）. The intracellular caspase-3 activity was determined by coloremetric assay. Concerntration of serum sTRAIL, anti-nucleosome antibodies （AnuA） and nucleosomes released from PBMC were assayed by sandwich enzyme linked immunosorbent assay （ELISA）. Results The expression levels of TRAIL mRNA was significantly higher in active SLE patients （0. 82 ± 0. 07 ） than inactive SLE patients（0.77±0.06）and healthy controls（0. 75 ±0. 05） （P 〈0. 05）. The Caspase-3 activity（A405） was significantly higher in active SLE patients （ 0.53 ± 0. 28 ） than inactive SLE patients （ 0. 39 ± 0. 23 ） and healthy controls （0. 35 ±0. 18） （P 〈0. 05）. The concerntration of nucleosome released from PBMC was significantly higher in acsTRAIL,Caspase-3 activity （ P 〈 0.05 ）, and between the Caspase-3 activity and levels of nucleosomes released from PBMC （ P 〈 0. 05 ）. There were positive correlation between TRAIL mRNA expression level and SLEDAI scores, anti-dsDNA, ESR, levels of serum AnuA concerntration; negative correlation between TRAILmRNA expression and complement C4, lymphocyte counts; significant positive correlation between serum sTRAIL concerntration and SLEDAI scores, negative correlation between sTRAIL concerntration and lymphocyte counts. There were positive correlation between levels of nucleosome and SLEDAI scores, dsDNA, ESR,levels of AnuA;negative correlation with complement C3, C4, lymphocyte counts. Conclusion The mean expression levels of TRAIL mRNA on PBMC and serum sTRAIL levels increase in SLE patients. TRAIL-mediated apoptosis may participate in the abnormal apoptosis of PBMC,increase nucleosome release and disease activity in SLE patients.