POEMS综合征患者高水平血清血管内皮生长因子的细胞来源及产生机制

发布时间：2018-05-11 00:12

本文选题：POEMS综合征 + VEGF　；参考：《北京协和医学院》2015年博士论文

【摘要】：背景和目的POEMS综合征是一种少见的单克隆浆细胞增殖性疾病,主要临床特点包括多发周围神经病、硬化性骨病和水负荷异常等。血管内皮生长因子(VEGF),作为一种调控血管新生和增强血管通透性的重要细胞因子,是该病诊断、病情监测和疗效评价的核心分子标志物。但是,目前关于POEMS综合征患者高水平血清VEGF的具体细胞来源,及其产生调控机制还缺乏系统的研究。材料与方法收集2014年2月至2015年4月就诊于北京协和医院的62例初治及46例其间完成治疗的POEMS综合征患者临床资料、血清和骨髓标本。采用ELISA法测定血清VEGF水平。通过实时荧光定量PCR和流式细胞学,分别在mRNA和蛋白水平测定骨髓内浆细胞表达VEGF的相对水平,并和血清VEGF水平进行相关性分析。利用流式细胞学,对骨髓内浆细胞亚群进行分析,并测定不同亚群细胞表达VEGF和白介素-6(IL-6)的水平。应用免疫组织化学法,检测患者骨髓活检组织中浆细胞分布、克隆性和VEGF及相关调控分子,包括IL-6和缺氧诱导因子1-α(HIF-1α)的表达情况。采用SPSS 22软件进行统计分析,当p0.05时认为存在统计学差异。结果初治POEMS综合征患者血清VEGF水平(中位值5958 pg/mL),显著高于其他疾病和健康对照(p0.001),具有明确的诊断价值(受试者工作曲线下面积0.988,p0.001),当选取2000 pg/mL作为临界值时,其诊断的特异性和敏感性分别为97.7%和91.9%。随治疗进行,血清VEGF水平逐渐下降(6程治疗后,中位值1184pg/mL, p 0.001;12程治疗后,中位值832 pg/mL, p 0.001). POEMS综合征患者骨髓内浆细胞VEGF在mRNA和蛋白水平上均高于对照,且随治疗逐步下降。重要的是,骨髓内浆细胞VEGF水平和血清VEGF水平有显著相关性(初治,rho= 0.33,p=0.01;治疗后,rho=0.53, p0.001)。在62例初治POEMS综合征患者中,51例经骨髓流式细胞学未检测到单克隆浆细胞,仅存在多克隆浆细胞。另11例患者(18%),骨髓中同时检测到单克隆和多克隆浆细胞,数量上以后者较多(41%vs.59%)。单克隆浆细胞VEGF水平与多克隆浆细胞类似(平均荧光强度2009 vs.2367,p=0.594),但IL-6水平显著高于后者(平均荧光强度1635 vs.865,p=0.006)。在46例骨髓活检组织中,POEMS综合征患者浆细胞呈现散在分布(25例,54%)或散在伴灶性分布(21例,46%)两种形式。其中散在浆细胞为多克隆,而灶性区域内同时存在单克隆和多克隆浆细胞,两者均表达VEGF和HIF-1α,而IL-6则主要为单克隆浆细胞表达。结论POEMS综合征患者骨髓内浆细胞是VEGF的主要来源。其中,单克隆浆细胞可能通过旁分泌IL-6,刺激多克隆浆细胞增生、产生VEGF。
[Abstract]:Background and objective POEMS syndrome is a rare monoclonal plasmacyte proliferative disease. The main clinical features include multiple peripheral neuropathy sclerosing osteopathy and abnormal water load. Vascular endothelial growth factor (VEGF), as an important cytokine to regulate angiogenesis and enhance vascular permeability, is the core molecular marker for diagnosis, disease monitoring and curative effect evaluation. However, there is no systematic study on the specific cell source and its production regulation mechanism of high level serum VEGF in patients with POEMS syndrome. Materials and methods Clinical data, serum and bone marrow samples of 62 patients with POEMS syndrome and 46 patients with POEMS syndrome who were admitted to Beijing Union Hospital from February 2014 to April 2015 were collected. Serum VEGF level was measured by ELISA method. The relative levels of VEGF expression in plasma cells of bone marrow were measured by real-time fluorescence quantitative PCR and flow cytometry, respectively, at the level of mRNA and protein, and the correlation between VEGF expression and serum VEGF level was analyzed. The plasma cell subsets in bone marrow were analyzed by flow cytometry, and the expression levels of VEGF and interleukin-6 (IL-6) in different subgroups were measured. The plasma cell distribution, clonality and the expression of VEGF and related regulatory molecules, including IL-6 and hypoxia inducible factor 1- 伪 (HIF-1 伪), in bone marrow biopsy tissues were detected by immunohistochemical method. SPSS 22 software was used for statistical analysis, when p0.05 there was a statistical difference. Results the serum VEGF level (median value 5958 PG / mL) in patients with newly diagnosed POEMS syndrome was significantly higher than that in other disease and healthy controls (P 0.001), and had a definite diagnostic value (the area under the operating curve of the subjects was 0.988g / ml). When 2000 pg/mL was selected as the critical value, The specificity and sensitivity of its diagnosis were 97.7% and 91.9%, respectively. With the treatment, the serum VEGF level decreased gradually after 6 weeks treatment, the median value was 1184pg / mL, p 0.001g / mL, the median value was 832pg / mL, p 0.001g / mL. Plasma cell VEGF in bone marrow of POEMS syndrome patients was higher than that of control group in mRNA and protein levels, and decreased gradually with treatment. Importantly, there was a significant correlation between plasma cell VEGF level in bone marrow and serum VEGF level (initial rhodia = 0.33 p0. 01, P 0. 001, P 0. 001, P 0. 001). In 62 cases of newly treated POEMS syndrome, 51 cases were not detected by bone marrow flow cytology, only polyclonal plasmacytes were found. In the other 11 patients, both monoclonal and polyclonal plasmacytes were detected in bone marrow. The VEGF level of monoclonal plasma cells was similar to that of polyclonal plasmacytes (mean fluorescence intensity was 2009 vs.2367, P < 0.05), but the IL-6 level was significantly higher than that of the latter (mean fluorescence intensity 1635 vs.865P0. 006). In 46 cases of bone marrow biopsy tissues, the plasma cells of patients with poems syndrome were in the form of scattered distribution (25 cases) or scattered with focal distribution (21 cases). The scattered plasmacytes were polyclonal, while in the focal region there were both monoclonal and polyclonal plasmacytes. Both expressed VEGF and HIF-1 伪, while IL-6 was mainly expressed by monoclonal plasmacytes. Conclusion Plasma cells in bone marrow of patients with POEMS syndrome are the main source of VEGF. Monoclonal plasma cells may stimulate the proliferation of polyclonal plasma cells by paracrine of IL-6 and produce VEGF.
【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R55

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