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MPO-ANCA相關(guān)性血管炎中性粒細(xì)胞CD35、CD55、MPO表達(dá)以及外周血、尿Ba水平的變化及意義

發(fā)布時(shí)間:2018-11-09 14:22
【摘要】:背景MPO-ANCA相關(guān)性血管炎(myeloperoxidase-specific antineutrophil cytoplasmic autoantibody associated vasculitis,MPO-AAV)是我國最常見的小血管炎類型之一,可表現(xiàn)為顯微鏡下多血管炎(Microscopic polyangiitis,MPA)、肉芽腫性多血管炎(Granulomatosis with polyangiitis,GPA)及嗜酸性肉芽腫性多血管炎(Eosinophilic granulomatosis with polyangiitis,EGPA)。由于這類疾病發(fā)病較隱匿、臨床表現(xiàn)多樣、進(jìn)展快,患者常因未及時(shí)診治而導(dǎo)致病情延誤甚至死亡。近年來,研究者們通過動(dòng)物實(shí)驗(yàn)、體外實(shí)驗(yàn)和臨床研究發(fā)現(xiàn)補(bǔ)體旁路途徑的激活在MPO-AAV的發(fā)病及病情進(jìn)展中可能起到了重要作用。補(bǔ)體系統(tǒng)的激活受到多種因素調(diào)控,如CD35、CD55。有研究報(bào)道顯示CD35、CD55在類風(fēng)濕關(guān)節(jié)炎(rheumatoid arthritis,RA)等自身免疫病中,對補(bǔ)體系統(tǒng)的調(diào)節(jié)起到了重要作用,而在MPO-AAV中,其與補(bǔ)體激活之間的關(guān)系尚未見研究報(bào)道。目的探討MPO-AAV患者中性粒細(xì)胞活性、補(bǔ)體旁路激活及調(diào)節(jié)劑水平的變化及其與臨床損害間的關(guān)系。方法收集40例活動(dòng)性MPO-AAV患者的全血、血清及尿液,并記錄患者的臨床損害情況及伯明翰血管炎活動(dòng)度積分(Birmingham Vasculitis Activity Score-version3,BVAS-V3)。另收集30例健康人全血、血清及尿液作為正常對照組。運(yùn)用流式細(xì)胞術(shù)(flow cytometry,FCM)分析MPO-ANCA陽性的患者和健康人外周血中性粒細(xì)胞CD35、CD55及MPO的表達(dá),同時(shí)運(yùn)用酶聯(lián)免疫吸附測定(enzyme-linked immunosorbent assay,ELISA)方法對患者和健康人血清及尿液中Ba水平進(jìn)行檢測,并探討CD35、CD55、MPO、Ba的水平與患者臨床損害間的關(guān)系。結(jié)果1.MPO-AAV患者組中性粒細(xì)胞CD35、CD55的表達(dá)水平均高于正常對照組(t=3.339,p=0.001;t=2.180,p=0.033)。MPO-AAV患者組MPO細(xì)胞內(nèi)表達(dá)水平低于正常對照組(t=-3.161,p=0.003);MPO-AAV患者組血清Ba濃度較對照組高(Z=-4.641,p0.001);2.MPO-AAV患者中性粒細(xì)胞CD35的平均熒光強(qiáng)度與血清Ba、尿液Ba呈正相關(guān)(r=0.336,p=0.034;r=0.324,p=0.041)。3.MPO-AAV患者中性粒細(xì)胞CD35的平均熒光強(qiáng)度與患者年齡(r=0.424,p=0.006)、C反應(yīng)蛋白(r=0.516,p=0.001)、補(bǔ)體C3(r=0.494,p=0.020)、外周血中性粒細(xì)胞計(jì)數(shù)(r=0.415,p=0.008)呈正相關(guān);CD55的平均熒光強(qiáng)度與患者年齡(r=0.514,p=0.001)、總BVAS(r=0.441,p=0.004)、CRP(r=0.377,p=0.020)、補(bǔ)體C4(r=0.445,p=0.038)、外周血中性粒細(xì)胞計(jì)數(shù)(r=0.485,p=0.001)、胸部損害BVAS(r=0.358,p=0.023)呈正相關(guān);MPO細(xì)胞內(nèi)表達(dá)的平均熒光強(qiáng)度與補(bǔ)體C4呈正相關(guān)(r=0.572,p=0.021);血清Ba濃度與患者病程呈負(fù)相關(guān)(r=-0.323,p=0.042);尿液中Ba與患者ESR(r=0.373,p=0.025)、CRP(r=0.399,p=0.013)、胸部BVAS(r=0.318,p=0.046)呈正相關(guān)。4.MPO-AAV患者中,有肺損害患者較無肺損害患者CD35和MPO的平均熒光強(qiáng)度更高(t=-2.373,p=0.023;t=-2.121,p=0.043);有腎損害患者較無腎損害患者CD55的MFI更高(t=-2.412,p=0.021)。結(jié)論1.補(bǔ)體激活抑制因子CD35、CD55在MPO-AAV患者中性粒細(xì)胞的表達(dá)是顯著增強(qiáng)的,可能減少中性粒細(xì)胞被激活的補(bǔ)體系統(tǒng)破壞,加重中性粒細(xì)胞參與的炎癥反應(yīng)。2.中性粒細(xì)胞CD35高表達(dá)的MPO-AAV患者肺臟可能更易受到累及;CD55高表達(dá)者腎臟可能更易受到累及。3.MPO-AAV患者中性粒細(xì)胞內(nèi)MPO的表達(dá)水平明顯降低,提示中性粒細(xì)胞脫顆粒中的MPO可能在MPO-AAV病情發(fā)生和發(fā)展中發(fā)揮更加重要的作用。4.MPO-AAV患者補(bǔ)體旁路激活明顯增加,但周圍血中Ba的升高水平與患者臨床損害嚴(yán)重性間無顯著相關(guān)性,患者尿中Ba水平的檢測,可能用于協(xié)助判斷MPO-AAV病情活動(dòng)性。
[Abstract]:Background MPO-ANCA-associated vascular inflammation (MPO-AAV) is one of the most common types of vasculitis in our country. It can be seen as a microscopic polyangitis (MPA), a granulomatous polyangitis (GPA) and an eosinophilic granuloma with polyangitis. EGPA). Because the disease of this kind of disease is more insidious, the clinical manifestations are various, the progress is fast, the patient often causes the disease to delay or even death due to the failure of timely diagnosis and treatment. In recent years, by animal experiments, in vitro experiments and clinical studies, the activation of the complement bypass pathway may play an important role in the pathogenesis of MPO-AAV and the progression of the disease. Activation of the complement system is regulated by a variety of factors, such as CD35, CD55. It is reported that CD35 and CD55 play an important role in the regulation of complement system in autoimmune diseases such as rheumatoid arthritis (RA), and the relationship between the complement activation in MPO-AAV has not been reported. Objective To study the changes of neutrophil activity, complement bypass activation and regulator level in patients with MPO-AAV and their relationship with clinical damage. Methods The whole blood, serum and urine of 40 patients with active MPO-AAV were collected and the clinical damage and the activity score of Birmingham Vasculitis Activity Score-version3, BVAS-V3 were recorded. The whole blood, serum and urine of 30 healthy people were collected as the normal control group. The expression of CD35, CD55 and MPO in peripheral blood of patients with MPO-ANCA positive and the expression of CD35, CD55 and MPO were analyzed by flow cytometry (FCM), and the levels of Ba in serum and urine of patients and healthy people were detected by enzyme-linked immunosorbent assay (ELISA), and CD35 was also discussed. The relationship between the level of CD55, MPO and Ba and the clinical damage of the patient. Results The expression of CD35 and CD55 in the patients with MPO-AAV was higher than that in the normal control group (t = 3.339, p = 0.001; t = 2.180, p = 0.033). The expression of MPO in the MPO-AAV group was lower than that in the control group (t =-3.161, p = 0.003); the serum Ba concentration in the MPO-AAV group was higher than that in the control group (Z =-4.641, p0.001); 2. The average fluorescence intensity of the neutrophils CD35 in the MPO-AAV group was positively correlated with the serum Ba and the urine Ba (r = 0.336, p = 0.034; r = 0.324, The mean fluorescence intensity of the C-reactive protein (r = 0. 516, p = 0. 001), the C-reactive protein (r = 0. 516, p = 0. 001), the complement C3 (r = 0.494, p = 0. 020), the peripheral blood neutrophil count (r = 0.415, p = 0. 008) was positively correlated with the patient's age (r = 0.415, p = 0. 008), and the mean fluorescence intensity of the CD55 was related to the patient's age (r = 0.514, p = 0.001), and the total BVAS (r = 0.441, p = 0. 004), CRP (r = 0.377, p = 0. 020), complement C4 (r = 0.445, p = 0.038), peripheral blood neutrophil count (r = 0.485, p = 0.001), chest damage BVAS (r = 0.358, p = 0.023), and positive correlation between the average fluorescence intensity and complement C4 in MPO cells (r = 0.572, p = 0.021); the serum Ba concentration was negatively correlated with the course of the patient (r =-0.323, In urine, Ba was positively correlated with ESR (r = 0.373, p = 0.025), CRP (r = 0.399, p = 0.013), breast BVAS (r = 0.318, p = 0.046), and the mean fluorescence intensity of CD35 and MPO in patients with lung damage in patients with lung impairment was higher in patients with lung impairment (t =-2.373, p = 0.023; t =-2.121, p = 0.043); The MFI of CD55 in patients with renal impairment was higher in patients with no renal impairment (t =-2.412, p = 0.021). Conclusion 1. The expression of the complement activation inhibitor CD35, CD55 in the MPO-AAV patient is significantly enhanced, and it is possible to reduce the complement system that is activated by the neutrophils and to aggravate the inflammatory response of the neutrophils. Neutrophil CD35 high-expressed MPO-AAV patient lung may be more susceptible to involvement; the CD55 high-expression kidney may be more susceptible to involvement. 3. The level of expression of MPO in the neutrophils of the MPO-AAV patient is significantly reduced, It was suggested that MPO might play a more important role in the pathogenesis and development of MPO-AAV. It may be used to assist in the determination of MPO-AAV disease activity.
【學(xué)位授予單位】:安徽醫(yī)科大學(xué)
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2015
【分類號】:R543

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