Lin Xu-Hong, Guo Jun-Ling, Wen Yu-Qing, Li Yu-Xia, Wei Dan-Dan, Yang Rui-Lin, Mu Xiao-Yun, Wang Hui-Chao
Department of Clinical Laboratory, Translational Medicine Center, Huaihe Hospital Affiliated to Henan University, Kaifeng 475000, China.
Department of Clinical Cardiology, Huaihe Hospital Affiliated to Henan University, Medical Sciences of Henan University, Kaifeng 475000, China.
Sheng Li Xue Bao. 2017 Apr 25;69(2):172-182.
The present study is designed to explore the role of plasma cells in the change of protein C system (PCS) in ulcerative colitis (UC). Dextran sulfate sodium (DSS, 4% in concentration) was used to induce mouse UC model. The plasma cells and the type of immune complex in colon were observed by immunofluorescence. The amount and type of plasma cells separated from colonic mucosal lamina propria were detected by flow cytometry using anti-CD54CD38 and IgA/M/G antibodies, respectively. After stimulation of macrophages by IgG type immune complex, TNF-α and IL-6 levels were evaluated by ELISA. After co-incubation of microvascular endothelial cells with TNF-α or IL-6, the expressions of endothelial protein C receptor (EPCR) and thrombomodulin (TM), and the activity of activated protein C (APC) were examined. As the results showed, the IgG type plasma cells infiltration and the quantity of IgG type immune complex were increased in DSS group in comparison with control group. After incubation with IgG type immune complex, the levels of TNF-α and IL-6 in the supernatant of macrophages were increased (P < 0.01) in a concentration-dependent manner. Meanwhile, after incubation with TNF-α or IL-6, the expressions of EPCR and TM in the microvascular endothelial cells were decreased (P < 0.05 or P < 0.01), while the activity of APC was reduced (P < 0.05 or P < 0.01). These results suggested that the quantity of IgG type plasma cells increases in UC and forms immune complexes, which affect the secretion of cytokines from macrophage, thereby affecting the function of endothelial cells and finally inhibiting PCS in UC. Therefore, plasma cell may be a novel target for the treatment of UC.
本研究旨在探讨浆细胞在溃疡性结肠炎（UC）中蛋白C系统（PCS）变化中的作用。采用硫酸葡聚糖钠（DSS，浓度为4%）诱导小鼠UC模型。通过免疫荧光观察结肠中的浆细胞和免疫复合物类型。分别使用抗CD54CD38和IgA/M/G抗体，通过流式细胞术检测从结肠黏膜固有层分离的浆细胞数量和类型。用IgG型免疫复合物刺激巨噬细胞后，通过酶联免疫吸附测定（ELISA）评估肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）水平。微血管内皮细胞与TNF-α或IL-6共同孵育后，检测内皮蛋白C受体（EPCR）和血栓调节蛋白（TM）的表达以及活化蛋白C（APC）的活性。结果显示，与对照组相比，DSS组IgG型浆细胞浸润及IgG型免疫复合物数量增加。用IgG型免疫复合物孵育后，巨噬细胞上清液中TNF-α和IL-6水平呈浓度依赖性升高（P<0.01）。同时，与TNF-α或IL-6孵育后，微血管内皮细胞中EPCR和TM的表达降低（P<0.05或P<0.01），而APC的活性降低（P<0.05或P<0.01）。这些结果表明，UC中IgG型浆细胞数量增加并形成免疫复合物，影响巨噬细胞细胞因子的分泌，从而影响内皮细胞功能，最终抑制UC中的PCS。因此，浆细胞可能是UC治疗的新靶点。
