IgG型浆细胞在溃疡性结肠炎小鼠蛋白C系统变化中的作用

发布时间：2019-03-30 14:14

【摘要】：本文旨在探讨IgG型浆细胞在溃疡性结肠炎(ulcerative colitis,UC)小鼠蛋白C系统(protein C system,PCS)变化中的作用。利用4%硫酸葡聚糖钠(dextran sulfate sodium,DSS)模拟小鼠UC,免疫荧光法观察结肠组织黏膜固有层浆细胞及免疫复合物IgA/M/G的类型,分离小鼠结肠组织黏膜固有层细胞,用抗CD38~+、CD54~+抗体双标、流式细胞术检测浆细胞数量,以抗IgA/M/G抗体标记,流式细胞术检测浆细胞类型;模拟IgG型免疫复合物刺激分离培养的巨噬细胞,ELISA法检测上清中促炎细胞因子TNF-α和IL-6的变化;流式细胞术检测TNF-α、IL-6对结肠黏膜微血管内皮细胞蛋白C受体(endothelial protein C receptor,EPCR)、血栓调节蛋白(thrombomodulin,TM)表达的影响,发色底物法检测TNF-α、IL-6对微血管内皮细胞激活的蛋白C(activated protein C,APC)活性的影响。结果显示:与对照组相比,DSS组小鼠结肠组织大量IgG型浆细胞浸润(P0.05),黏膜固有层IgG型免疫复合物水平显著升高;分离培养的巨噬细胞与模拟IgG型免疫复合物共孵育后,上清中炎性细胞因子TNF-α和IL-6明显增高(P0.01);同时TNF-α或IL-6与小鼠结肠黏膜微血管内皮细胞共孵育后,内皮细胞表达EPCR、TM的能力均有所降低(P0.05或P0.01),其APC活性明显降低(P0.05或P0.01)。以上结果提示,UC时IgG型浆细胞数量增加,并通过形成免疫复合物,从而影响巨噬细胞分泌促炎细胞因子,进而影响血管内皮细胞功能,抑制PCS。浆细胞有望成为治疗UC的新靶点。
[Abstract]:The aim of this study was to investigate the role of IgG type plasma cells in the changes of protein C system (protein C system,PCS) in ulcerative colitis (ulcerative colitis,UC) mice. The types of plasma cells and immune complex IgA/M/G in colonic mucosa of mice were observed by 4% sodium dextran sulfate (dextran sulfate sodium,DSS) imitated by UC, immunofluorescence. The cells of the lamina propria of colonic mucosa were isolated and anti-CD38~ was used. CD54~ antibody double labeling, flow cytometry to detect the number of plasma cells, anti-IgA/M/G antibody labeling, flow cytometry to detect the type of plasma cells; Macrophage was isolated and cultured by simulating IgG immune complex. The changes of pro-inflammatory cytokines TNF- 伪 and IL-6 in supernatant were detected by ELISA method. The effects of TNF- 伪 and IL-6 on the expression of protein C receptor (endothelial protein C receptor,EPCR and thrombomodulin (thrombomodulin,TM) in colonic microvascular endothelial cells were detected by flow cytometry, and the expression of TNF- 伪 was detected by chromogenic substrate method. The effect of IL-6 on the activation of C (activated protein C, APCs in microvascular endothelial cells. The results showed that compared with the control group, a large number of IgG-type plasma cells infiltrated in the colon of DSS group (P0.05), and the level of IgG-type immune complex in the mucosal lamina propria was significantly higher than that in the control group. The levels of TNF- 伪 and IL-6 in the supernatant of cultured macrophages were significantly increased after co-incubation with simulated IgG immune complexes (P0.01). At the same time, when TNF- 伪 or IL-6 were co-incubated with mouse colonic microvascular endothelial cells, the ability of EPCR,TM expression was decreased (P0.05 or P0.01), and the activity of APC was significantly decreased (P0.05 or P0.01). These results suggest that the number of IgG type plasmacytes increases in UC, and the immune complexes are formed to affect the secretion of pro inflammatory cytokines by macrophages, thereby affecting the function of vascular endothelial cells and inhibiting PCS.. Plasma cells are expected to be a new target for the treatment of UC.
【作者单位】：河南大学淮河医院检验科转化医学中心;河南大学淮河医院心内科河南大学医学院;河南大学淮河医院甲状腺乳腺外科河南大学医学院;河南大学第一附属医院肾内科;
【基金】：supported by the National Natural Science Foundation of China(No.81500430,U1304802) the Key Projects of Science and Technology from the Education Department of Henan Province,China(No.17A320019) the Research Foundation of Henan University,Henan Province,China(No.2012YBWT04,yqpy20140012,2012YBZR020)
【分类号】：R574.62

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