【作者】 李劲频；

【导师】 莫雪安；

【作者基本信息】 广西医科大学 ， 神经病学， 2002， 硕士

【摘要】 目的：(1)研究重症肌无力(MG)患者CD40配体(CD40L)的表达，探讨CD40L在MG中的作用。(2)探讨MG患者CD40L表达的信号传导途径。 方法：研究对象为25例MG患者，分为急性期组13例，非急性期组12例，设正常对照组15例。分离血中单个核细胞，用流式细胞仪检测CD40L阳性细胞率，应用RT—PCR方法检测单个核细胞CD40L mRNA的表达。再分组用刺激剂刺激进行单个核细胞培养：①PMA组：即用PMA和A23187刺激；②PHA组：即用PHA刺激。培养后用流式细胞仪检测CD40L阳性细胞率及RT—PCR法检测单个核细胞CD40LmRNA，并与刺激前作比较，同时还进行急性期组、非急性期组和正常对照组之间的比较。急性期组先加PKC抑制剂BLM后再加PHA刺激培养(BLM组)，然后检测CD40L阳性细胞率及CD40LmRNA(方法如前述)，并与PMA组、PHA组及纯培养组作比较分析。 结果：(1)MG急性期组CD40L阳性细胞率和CD40L mRNA均比正常对照组及非急性期组显著增高(均为P＜0.01)，而非急性期组CD40L阳性细胞率和CD40LmRNA与正常对照组相比无显著性差异(P＞0.05)；(2)经PMA或PHA刺激后， 广西医科大学硕士研究生学位论文 不但急性期组 CD40L阳性细胞率和 CD40 Lm叫A仍比非急性 期组和正常对照组显著增高（均为P＜0．01卜而且非急性期组上 述指标也比正常对照组显著增高呼 刀1）；门）MG急性期 BLM组 CD40L阳性细胞率和 CD40L mRNA与纯培养组无明显 差别（Ptoo．05），而显著低 于PMA组和P枷组（Pwto．01）。 结论：（）CD40L的高表达是MG发生和加重的因素八2） 在MG中，CD40L的表达可能是依赖于PKC活性介导的T细 胞活化途径。更多还原

【Abstract】 Objectives: (l)To investigate the role of CD40 and CD40 ligand in the pathogenesis of myasthenia gravis (MG) , the CD40L expression on the peripheral blood mononuclear cells of patients with MG was tested. (2) To investigate the signal transduction pathway involved in CD40L expression of patients with MG.Methods: 25 patients with MG included 13 acute patients and 12 non-acute patients. 15 healthy people was acted as control group. Blood of patients and normal controls was collected. Mononuclear cells in peripheral blood(PBMC) were isolated and cultured in the medium(RPMI-1640) supplemented with PMA and A23187 or PHA. After 6 hours, cell samples were collected and CD40L expression was tested by flow cytometry and reverse transcription-polymerase chain reaction(RT-RCR) compared with that of them before activation. Except for PMA and A23187 or PHA treatment, a specific inhibitor of PKC, bisindolylmaleimide(BLM),was added to the cells before exposure to PHA in patients with acute MG. Meanwhile, the medium without activator in PBMC cultures was used in acute MG .Cells indifferent culture medium were collected and CD40L expression was tested .Results: (1) CD40L expression on PBMC in patients with acute MG was increased markedly compared with that of patients with non-acute MG and normal controls before activation, but there were no significantly statistics differences between patients with non-acute MG and normal controls. (2)CD40L expression of patients with acute MG and non-acute MG were increased significantly in contrast to that of normal controls after activation with PHA or PMA CD40L expression of patients with acute MG still increased markedly compared with that of patients with non-acute MG. (3)CD40L expression of patient with acute MG after activation with PHA was not different from that after activation with PMA, but there was increased markedly in contrast to that of the status of medium without activator. CD40L expression in the presence of the PKC inhibitor was no significantly difference compared with that of the status of medium without activator.Conclusions: (1) Hyperexpression of CD40L is correlated with the clinical progression of MG. (2) CD40L expression in MG is mediated by PKC activity.更多还原