目的 检测自噬相关蛋白Beclin 1和LC3在多发性硬化(MS)和视神经脊髓炎(NMO)患者急性期治疗前、后及复发型急性期间外周血单个核细胞(PBMC)中的表达水平并探讨其意义.方法 所有研究对象均为2012年10月至2014年4月在苏州大学附属第一医院住院、门诊患者及健康体检者,采用密度-梯度离心法分离MS、NMO急性期患者(治疗前、后)与正常对照组的外周血中PBMC,应用Western印迹技术检测Beclin 1和LC3的表达水平.同时,分别随访复发缓解型MS(RRMS)和复发型NMO(RNMO)患者,对比两次急性期Beclin 1和LC3的表达变化情况.结果 (1)与对照组相比,急性期MS和NMO患者外周血PBMC中Beclin 1的表达降低,LC3表达增高(P<0.01).(2)MS、NMO患者经过治疗后对比治疗前Beclin 1表达明显增高,LC3表达则明显降低(P<0.01).(3)然而随访发现RRMS和RNMO患者复发期较前次发作期Beclin 1的表达水平降低与LC3的表达增高均更加显著(P<0.01).结论 急性期MS和NMO患者外周血PBMC中自噬水平增高,经治疗后自噬水平下降;然而在RRMS和RNMO患者复发期自噬水平进一步增高.这表明自噬水平的高低可能与MS和NMO的发生及复发相关。
Objective To explore the expression and significance of Beclin 1 and LC3 in peripheral blood mononuclear cells (PBMCs) of patients with multiple sclerosis (MS) and neuromyelitis optica (NMO) before and after treatment during acute and recurrent acute phases.Methods All outpatients,inpatients and healthy controls were recruited from our hospital from October 2012 to April 2014.During acute phase,PBMCs from patients with multiple sclerosis (MS) and neuromyelitis optica (NMO) (pre and post-treatment) were immediately isolated by Ficoll-Hypaque density gradient centrifugation.And the expressions of Beclin 1 and LC3 were detected by Western blot.And relapsing-remitting MS (RRMS) and relapsing NMO (RNMO) patients were followed up and the expressions of Beclin 1 and LC3 proteins compared during two acute phases.Results Compared with normal controls,the expression of Beclin 1 in PBMCs from acute phase of MS and NMO patients decreased while the expression of LC3 increased.During acute phase,the expression of Beclin 1 significantly increased after treatment in MS and NMO patients compared with pre-treatment,but the expression of LC3 significantly decreased.The expression of Beclin1 obviously decreased in RRMS and recurrent RNMO compared with the previous period,but the expression of LC3 significantly increased.Conclusion The autophagy level increases in PBMC from MS and NMO patients during acute phase.However it decreases after treatment.However,the autophagy levels significantly increase in RRMS and RNMO.And enhanced autophagy may play its role in the pathogenesis of MS and NMO.