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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(P<0.05),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(P<0.01);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(P<0.05或P<0.01),其APC活性明显降低(P<0.05或P<0.01)。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。
This article aims to investigate the role of IgG-type plasma cells in the changes of protein C system (PCS) in ulcerative colitis (UC) mice. The UC of mice was simulated with 4% dextran sulfate sodium (DSS), and the type of IgA / M / G in plasma lamina propria and mucosa lamina propria of colon were observed by immunofluorescence. The colon mucosa of mice was isolated The number of plasma cells was detected by flow cytometry, anti-IgA / M / G antibody labeling, and the type of plasma cells by flow cytometry. The immune response of IgG-like immune complexes The cultured macrophages were stimulated and the changes of pro-inflammatory cytokines TNF-α and IL-6 in supernatants were detected by ELISA. The expressions of TNF-α and IL-6 in colonic mucosa microvascular endothelial cells were detected by flow cytometry (EPCs) and thrombomodulin (TM) expression were detected by enzyme-linked immunosorbent assay (ELISA). The activated protein C (APC) activity of TNF-αand IL-6 in microvascular endothelial cells Impact. The results showed that compared with the control group, the colon tissue of mice in the DSS group had a large number of IgG-type plasma cells infiltrating (P <0.05), and the level of IgG-type immune complex in the lamina propria of the DSS group was significantly increased. Compared with the control group, After incubation with immunocomplex, the levels of TNF-α and IL-6 in supernatants were significantly increased (P <0.01). After co-incubation of TNF-α or IL-6 with mouse colonic mucosal microvascular endothelial cells, The ability of cells to express EPCR and TM decreased (P <0.05 or P <0.01), and APC activity decreased significantly (P <0.05 or P <0.01). The above results suggest that the number of IgG-type plasma cells increases at UC, and through the formation of immune complexes, thereby affecting the secretion of pro-inflammatory cytokines macrophages, thus affecting vascular endothelial cell function, inhibit PCS. Plasma cells are expected to become new targets for the treatment of UC.