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目的 探讨狼疮肾炎 (LN)病人单核细胞凋亡加速的机制。方法 对 17例LN病人进行观察 ,以 15名正常人为对照。采用流式细胞仪检测单核细胞Fas和FasLigand (FasL)的表达 ;酶联免疫吸附法 (ELISA)测定血浆中可溶性FasLigand (sFasL)的水平 ;单核细胞与重组人FasL共孵育 ,碘化丙啶染色、流式细胞仪检测单核细胞凋亡 ,四甲基偶氮唑蓝 (MTT)法观察单核细胞体外存活率。结果 LN病人单核细胞Fas的表达较正常人明显上调 (P <0 0 5 ) ;处于狼疮活动期的LN病人与狼疮静止期的LN病人之间单核细胞Fas的表达差异无显著性 (P >0 0 5 )。正常人和LN病人的单核细胞表面均未检测出FasL的表达。正常人血浆中未能检测出sFasL ,但LN病人血浆存在有sFasL ;sFasL的水平在狼疮活动期和静止期的LN病人之间差异无显著性 (P >0 0 5 )。单核细胞与重组人FasL在体外共同培养 2h后 ,LN病人单核细胞的凋亡较正常人明显增高 (P <0 0 5 ) ,单核细胞与重组人FasL在体外共同培养 6h后 ,LN病人单核细胞的存活率较正常人明显低下 (P <0 0 5 )。结论 LN病人单核细胞表面功能性Fas表达上调 ,血浆中存在sFasL ,这可能是LN病人单核细胞凋亡加速的原因之一
Objective To investigate the mechanism of accelerated apoptosis of monocytes in patients with lupus nephritis (LN). Methods 17 cases of LN patients were observed, with 15 normal controls. The expression of Fas and FasLigand (FasL) in monocytes was detected by flow cytometry. The levels of soluble FasLigand (sFasL) in plasma were detected by enzyme-linked immunosorbent assay (ELISA). The mononuclear cells were incubated with recombinant human FasL, The apoptosis of monocytes was detected by flow cytometry. The survival rate of monocytes was observed by MTT assay. Results The expression of Fas in monocytes in LN patients was significantly higher than that in normal subjects (P <0.05). There was no significant difference in the expression of Fas in monocytes between LN patients with active lupus and those with inactive lupus > 0 0 5). No expression of FasL was detected on the mononuclear cells of normal and LN patients. No sFasL was detected in normal plasma, but there was sFasL in plasma of patients with LN. The level of sFasL was not significantly different between lupus active and quiescent LN patients (P> 0.05). Monocytes and recombinant human FasL co-cultured in vitro 2h, LN patients with monocytes apoptosis was significantly higher than normal (P <0 05), monocytes and recombinant human FasL co-cultured in vitro 6h, LN The survival rate of patients with monocytes was significantly lower than normal (P <0 05). Conclusions The expression of functional Fas on the surface of mononuclear cells in patients with LN is up-regulated and sFasL is present in plasma, which may be one of the reasons for the accelerated apoptosis of mononuclear cells in patients with LN