【摘要】：本文旨在探討IgG型漿細(xì)胞在潰瘍性結(jié)腸炎(ulcerative colitis,UC)小鼠蛋白C系統(tǒng)(protein C system,PCS)變化中的作用。利用4%硫酸葡聚糖鈉(dextran sulfate sodium,DSS)模擬小鼠UC,免疫熒光法觀察結(jié)腸組織黏膜固有層漿細(xì)胞及免疫復(fù)合物IgA/M/G的類型,分離小鼠結(jié)腸組織黏膜固有層細(xì)胞,用抗CD38~+、CD54~+抗體雙標(biāo)、流式細(xì)胞術(shù)檢測(cè)漿細(xì)胞數(shù)量,以抗IgA/M/G抗體標(biāo)記,流式細(xì)胞術(shù)檢測(cè)漿細(xì)胞類型;模擬IgG型免疫復(fù)合物刺激分離培養(yǎng)的巨噬細(xì)胞,ELISA法檢測(cè)上清中促炎細(xì)胞因子TNF-α和IL-6的變化;流式細(xì)胞術(shù)檢測(cè)TNF-α、IL-6對(duì)結(jié)腸黏膜微血管內(nèi)皮細(xì)胞蛋白C受體(endothelial protein C receptor,EPCR)、血栓調(diào)節(jié)蛋白(thrombomodulin,TM)表達(dá)的影響,發(fā)色底物法檢測(cè)TNF-α、IL-6對(duì)微血管內(nèi)皮細(xì)胞激活的蛋白C(activated protein C,APC)活性的影響。結(jié)果顯示:與對(duì)照組相比,DSS組小鼠結(jié)腸組織大量IgG型漿細(xì)胞浸潤(rùn)(P0.05),黏膜固有層IgG型免疫復(fù)合物水平顯著升高;分離培養(yǎng)的巨噬細(xì)胞與模擬IgG型免疫復(fù)合物共孵育后,上清中炎性細(xì)胞因子TNF-α和IL-6明顯增高(P0.01);同時(shí)TNF-α或IL-6與小鼠結(jié)腸黏膜微血管內(nèi)皮細(xì)胞共孵育后,內(nèi)皮細(xì)胞表達(dá)EPCR、TM的能力均有所降低(P0.05或P0.01),其APC活性明顯降低(P0.05或P0.01)。以上結(jié)果提示,UC時(shí)IgG型漿細(xì)胞數(shù)量增加,并通過(guò)形成免疫復(fù)合物,從而影響巨噬細(xì)胞分泌促炎細(xì)胞因子,進(jìn)而影響血管內(nèi)皮細(xì)胞功能,抑制PCS。漿細(xì)胞有望成為治療UC的新靶點(diǎn)。
[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- 偽 and IL-6 in supernatant were detected by ELISA method. The effects of TNF- 偽 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- 偽 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- 偽 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- 偽 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.
【作者單位】： 河南大學(xué)淮河醫(yī)院檢驗(yàn)科 轉(zhuǎn)化醫(yī)學(xué)中心;河南大學(xué)淮河醫(yī)院心內(nèi)科 河南大學(xué)醫(yī)學(xué)院;河南大學(xué)淮河醫(yī)院甲狀腺乳腺外科 河南大學(xué)醫(yī)學(xué)院;河南大學(xué)第一附屬醫(yī)院腎內(nèi)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)
【分類號(hào)】：R574.62

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