抗中性粒细胞胞浆抗体诱导FcγRⅡa升高与血管炎相关细胞因子的关系

Relationship of FcγRⅡa expression induced by antineutrophil cytoplasmic antibody with cytokines

目的探讨抗中性粒细胞胞浆抗体是否通过影响中性粒细胞膜上FcγRⅡa的表达参与疾病进程。方法用不同浓度的抗中性粒细胞胞浆抗体相关血管炎活动期血清,分别是抗髓过氧化物酶抗体及抗蛋白酶3抗体阳性血清刺激健康人中性粒细胞,以健康人血清刺激中性粒细胞作为对照组,用流式细胞术分析血液中FcγRⅡa的荧光强度,Western blot分析FcγRⅡa的表达,鲁米诺化学发光方法检测中性粒细胞中氧自由基生成,用酶联免疫吸附试验检测血清细胞因子TNF-α、IL-6的水平及血清中抗髓过氧化物酶抗体及抗蛋白酶3抗体水平。结果伴随血清浓度的降低,FcγRⅡa荧光强度、FcγRⅡa蛋白的表达、中性粒细胞中氧自由基及其分泌的细胞因子TNF-α、IL-6水平下降(P<0.05)。结论抗髓过氧化物酶抗体及抗蛋白酶3抗体可能均通过FcγRⅡa影响氧自由基生成及细胞因子TNF-α、IL-6的水平,进而导致血管炎的发生。

Objective To investigate antineutrophil cytoplasmic antibody resulting in antineutrophil cytoplasmic antibody-associated vasculitides contributed by the expression of FcγRⅡa in the cell membrane. Methods Hu- man neutrophils were stimulated by different serum concentrations（ anti-myeloperoxidase antibody positive and anti-proteinase 3 positive） , healthy human serum as control. Flow eytometry was used to detect the expression of FcγRⅡa on the neutrophil membrane. FcγRⅡa was analyzed by Western blots. Reactive oxygen species （ROS） production was measured by chemiluminescence technique in the presence of 10 mmol/L luminol. ELISA was applied to detect the level of serum cytokines TNF-α, IL-6, anti-myeloperoxidase antibody and an- ti-proteinase 3. Results With the decrease of serum concentration, fluorescence intensity of FcγRⅡa expres- sion, expression of FcγRⅡa, ROS generation and secretion of cytokines TNF-α and IL- 6 reduced （ P 〈 0.05）. Conclusions Anti-myeloperoxidase antibody and anti-proteinase 3 antibody play a role in mediating ROS, TNF-α and IL-6 levels through FcγRⅡa, and thus induce the generation of antineutrophil cytoplasmic antibody associated vaseulitis.