摘要
目的通过小鼠腹腔注射5-乙炔基-2′脱氧尿嘧啶核苷（5-ethynyl-2′-deoxyuridine, EdU）标记骨髓中处于增殖状态的粒细胞，以期了解炎症状态下小鼠骨髓中的不同分化阶段的粒细胞生成的变化。方法小鼠腹腔注射热灭活的大肠杆菌（HI E．coli）建立急性腹腔炎症模型，采用EdU掺入法，随后24、48、72、96 h将小鼠处死并取小鼠全骨髓细胞，应用流式抗体lineage、CD34、c-Kit、Ly6G标记粒细胞谱系中的原始粒细胞、早幼粒细胞、中幼粒细胞、晚幼粒细胞以及杆状和分叶核细胞，瑞氏染色观察五群细胞的形态，结合EdU标记，明确炎症状态下不同分化阶段粒细胞的增殖水平的变化。结果小鼠腹腔注射HI E．coli 24、48、72、96 h后，与生理状态下相比，急性炎症状态下原始粒细胞EdU阳性细胞数目在24 h增加了10.0%，在48 h和72 h分别降低了75.0%和23.0%。早幼粒细胞EdU阳性细胞数目在24、48、72、96 h呈现下降的趋势，分别下降了23.0%、54.5%、64.3%和77.8%。中幼粒细胞的EdU阳性细胞数目在24、48 h之间呈现增加的趋势，分别增加了60.0%和10.0%，在72、96 h呈现下降的趋势，分别下降了80.0%和90.0%。晚幼粒细胞EdU的阳性细胞数目在24 h增加了50.0%，在48、72、96 h之间呈现下降的趋势，分别降低了33.3%、61.5%和66.7%。杆状和分叶核粒细胞EdU的阳性细胞数目在24 h增加了14.0%，在48、72、96 h呈现降低的趋势，分别下降了50.0%、77.8%和88.0%。结论在建立的HI E．coli诱导的急性腹膜炎模型发生24 h后，出现典型的急性髓系造血，即粒系前体细胞增殖加快，以中幼粒细胞以及晚幼粒细胞为主，导致成熟中性粒数目增多，可能多为趋化至炎症部位的成熟粒细胞。
Objective To label granulocytes in a state of differentiation in mouse bone marrow （BM） with EdU （5-ethynyl-2′-deoxyuridine） for further understanding the changes in granulocyte production at different stages of differentiation during inflammation.MethodsC57BL/6 mice were intraperitoneally （i.p.） injected with EdU and heat-inactivated Escherichia coli （HI E. coli）. BM cells were harvested at different time points after HI E. coli injection and then stained with fluorescent-conjugated antibodies （Abs）. Myeloblasts, promyelocytes, myelocytes, metamyelocytes and band and segmented neutrophils were identified by fluorescence-activated cell sorting （FACS）. The percentage of EdU-positive cells in each population was recorded.ResultsThe percentage of EdU-positive myeloblasts in mice increased by 10.0% at 24 h after intraperitoneal injection with HI E. coli, but decreased by 75.0% and 23.0% at 48 h and 72 h, respectively. The percentage of EdU-positive promyelocytes declined by 23.0%, 54.5%, 64.3% and 77.8% at 24 h, 48 h, 72 h and 96 h, respectively. The percentage of EdU-positive myelocytes increased by 60.0% and 10.0% at 24 h and 48 h, but decreased by 80.0% and 90.0% at 72 h and 96 h. The percentage of EdU-positive metamyelocytes increased by 50.0% at 24 h, but decreased by 33.3%, 61.5% and 66.7% at 48 h, 72 h and 96 h. The percentage of EdU-positive band and segmented cells increased by 14.0% at 24 h, but decreased by 50.0%, 77.8% and 88.0% at 48 h, 72 h and 96 h.ConclusionEmergency granulopoiesis occurred 24 h after the establishment of HI E. coli-induced model of acute peritonitis, which meant that the proliferation of myeloid precursor cells, especially that of myelocytes and metamyelocytes, was accelerated and resulted in increasing number of mature neutrophils immigrating to sites of inflammation.
作者
任仟
张潇予
郭荣霞
谢忻琰
张苏东
谢雪梅
范玉萍
王永荣
张存玲
王彤
刘飞
刘鹏
许元富
罗鸿博
Ren Qian;Zhang Xiaoyu;Guo Rongxia;Xie Xinyan;Zhang Sudong;Xie Xuemei;Fan Yuping;Wang Yongrong;Zhang Cunling;Wang Tong;Liu Fei;Liu Peng;Xu Yuanfu;Luo Hongbo(State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin 300020, Chin)
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2018年第4期254-259,共6页
Chinese Journal of Microbiology and Immunology
基金
国家重点基础研究发展计划（973计划）（2015CB964903）
中国医学科学院医学科学创新基金（2016-12M-1-003
2017-12M-1-015）
国家自然科学基金（31471116,81600083
31700783）
“协和青年基金”和“中央高校基本科研业务费专项资金”（2017310023,2017P＇131033）
关键词
EDU
粒细胞
炎症状态
增殖
EdU
Granulocyte
Inflammation
Proliferation
