CAJ | 학술논문

目的观察活动性系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMC)的自噬现象并检测相关基因的表达水平. 方法以初诊及新发作的活动性SLE患者20例为病例组,类风湿关节炎(RA)患者10例和健康志愿者10名分别作为对照组.密度梯度离心法分离PBMC,并利用贴壁法除去单核细胞.运用透射电镜(TEM)观察细胞的超微结构,半定量反转录-聚合酶链反应(RT-PCR)及实时定量RT-PCR检测各组PBMC中Beclin1、微管相关蛋白1轻链3(MAPLC3)mRNA的表达水平. 结果 TEM发现活动性SLE患者PBMC中存在自噬现象;Beclinl和MAPLC3 mRNA在活动性SLE患者表达量均增高,且分别较RA组和健康对照组表达差异有统计学意义. 结论活动性SLE患者外周血PBMC存在自噬现象,且细胞自噬相关基因表达较健康对照组和RA组增强,这可能与SLE的发生、发展相关.
목적 관찰활동성계통성홍반랑창(SLE)환자외주혈단개핵세포(PBMC)적자서현상병검측상관기인적표체수평. 방법 이초진급신발작적활동성SLE환자20례위병례조,류풍습관절염(RA)환자10례화건강지원자10명분별작위대조조.밀도제도리심법분리PBMC,병이용첩벽법제거단핵세포.운용투사전경(TEM)관찰세포적초미결구,반정량반전록-취합매련반응(RT-PCR)급실시정량RT-PCR검측각조PBMC중Beclin1、미관상관단백1경련3(MAPLC3)mRNA적표체수평. 결과 TEM발현활동성SLE환자PBMC중존재자서현상;Beclinl화MAPLC3 mRNA재활동성SLE환자표체량균증고,차분별교RA조화건강대조조표체차이유통계학의의. 결론 활동성SLE환자외주혈PBMC존재자서현상,차세포자서상관기인표체교건강대조조화RA조증강,저가능여SLE적발생、발전상관.
Objective To investigate the autophagy and the expression of its related genes in the peripheral blood mononuclear cells (PBMCs) of active systemic lupus erythematosus (SLE) patients.Methods Patients with newly onset or recently-diagnosed SLE (n=20) were enrolled.RA patients (n=10) and healthy blood donors (n=10) were used as controls.PBMCs from all subjects were immediately isolated by Ficoll-Hypaque density gradient centrifugation.And then monocytes were removed by wall sticking method.The morphology was observed using transmission electron microscopy (TEM).Messenger RNA (mRNA) expression of Beclin 1 and microtubule-associated protein 1-light chain 3 (MAPLC3) were determined by reverse transcription polymerase chain reaction (RT-PCR) and real-time quantitative RT-PCR respectively.Results TEM showed autophagic phenomenon in PBMCs from active SLE.On the mRNA level,expression of Beclin 1 and LC3 was significantly increased in fresh isolated SLE cells as compared with RA or healthy donor's PBMCs.Conclusion Based on these results,we can conclude that autophagy occurs in active SLE and the expression of its related genes is significantly higher in active SLE than in RA or normal controls.The enhanced autophagy may indicate its role in the pathogenesis of SLE.