嗜中性粒细胞是人抵抗素表达的主要细胞  

Expression of the Human Resistin in Neutrophil

作　　者：高振秋[1] 张巧[1] 徐彤[2] 田辉[1] 任桂萍[1] 李璐[1] 刘向宇[1] 李德山[1] 

机构地区：[1]东北农业大学生命科学学院生物制药教研室,哈尔滨150030 [2]黑龙江省医院检验科,哈尔滨150036

出　　处：《中国生物化学与分子生物学报》2011年第1期48-54,共7页Chinese Journal of Biochemistry and Molecular Biology

基　　金：黑龙江省教育厅项目(No.11521022;2007);哈尔滨市科技创新人才研究专项资金(No.2008RFQXS017)~~

摘　　要：抵抗素(resistin)是小鼠白色脂肪组织大量表达的富含半胱氨酸的分泌型蛋白.近年研究发现,人与啮齿类动物的抵抗素组织表达分布存在很大差异.小鼠抵抗素主要在白色脂肪组织表达,而人抵抗素主要在单核细胞/巨噬细胞表达,且在骨髓组织中大量表达,但目前骨髓中的细胞定位还不清楚.本研究的目的是明确成人骨髓及外周血白细胞中抵抗素表达细胞的类型.免疫荧光法检测骨髓中抵抗素表达细胞,结果显示,抵抗素主要表达在细胞核呈杆状和分叶核状的成熟粒细胞中,其中杆状核粒细胞表达较高,分叶核粒细胞表达减弱.Anti-hresistin IgG-Biotin-PE单色荧光流式细胞术分选外周血白细胞中抵抗素表达细胞后经瑞氏化学染色,结果显示,抵抗素表达细胞主要为杆状和分叶核状的嗜中性粒细胞,还有少量嗜酸性粒细胞,且抵抗素蛋白分布在细胞质中.RT-qPCR结果在RNA水平上证明,人抵抗素在嗜中性粒细胞中大量表达.Anti-hresistin IgG-FITC和anti-HNL IgG-Biotin-PE双色荧光流式细胞术进一步证明,抵抗素的主要表达细胞为成熟的嗜中性粒细胞.嗜中性粒细胞在机体免疫防御中起重要作用,人骨髓及外周血中抵抗素主要在成熟嗜中性粒细胞中表达,这一研究结论为人抵抗素与炎症反应的关联性及其功能的进一步研究奠定了基础.Resistin is a secreted protein with high level of Cys expressed from mouse white adiposetissue. Recently,many studies show that expression patterns of human and mouse resistin are different.Mouse resistin is primarily expressed in white adipose tissue,whereas human resistin is highly expressedin PBMCs /macrophages and bone marrow. But the cellular localization of the protein in bone marrow isnot clear. In the present study,in order to identify the resistin-expressing cells in bone marrow,wedetected the expression of resistin in bone marrow by immunofluorescence. The result showed that humanresistin was dominantly expressed in mature neutrophils with rod and segment-shaped nuclei,and the expression level was higher in the cells with rod-shaped nuclei than the segment-shaped ones. Anti-hresistin IgG-Biotin-PE stained cells were sorted and analyzed by Wright dye staining. The results showedthat human resistin was primarily stored in neutrophils and less in eosinophils of peripheral WBCs.Moreover,human resistin was present in cytoplasm. RT-qPCR analysis revealed that monocytes andneutrophils were the primary cells of the human peripheral WBCs. In order to confirm the cell type thatexpressed resistin,we detected bone marrow cells and peripheral WBCs expressing both HNL and resistinby two-color fluorescence analysis,and the results indicated that the dominant resistin-expressing cellswere neutrophils. Hence,we conclude that the majority of the resistin-expressing cells in human areneutrophils,which sheds light on the relationship between inflammation and the expression of humanresisitin.

关 键 词：人抵抗素 骨髓 外周血 嗜中性粒细胞 

分 类 号：Q78[生物学—分子生物学]