脓毒症患者中性粒细胞内钙水平下降影响细胞迁移研究  

Impact of intracellular calcium reduction on neutrophil migration in sepsis patients

作　　者：齐妍[1] 王闰楠 王贺元[3] 柳忠辉[1] Qi Yan;Wang Runnan;Wang Heyuan;Liu Zhonghui(Department of Immunology,College of Basic Medical Sciences,Jilin University,Changchun 130021,China;Department of Genetics,College of Basic Medical Sciences,Jilin University,Changchun 130021,China;Department of Endocrinology and Metabolism,the First Hospital of Jilin University,Changchun 130021,China)

机构地区：[1]吉林大学基础医学院免疫学系,长春130021 [2]吉林大学基础医学院遗传学系,长春130021 [3]吉林大学第一临床医院内分泌代谢科,长春130021

出　　处：《中华微生物学和免疫学杂志》2024年第3期212-216,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金(31871510);吉林省卫生健康委科学技术项目(2021JL033)。

摘　　要：目的探究细胞内钙水平与脓毒症患者中性粒细胞迁移功能障碍的关系。方法纳入吉林大学第一医院2017年12月—2018年9月临床诊断为脓毒症的21例患者血样,另外选取20名健康人作为对照组。采用磁珠分选试剂盒分离健康对照及脓毒症患者外周血中性粒细胞,多通道微流控芯片技术检测中性粒细胞的趋化迁移能力。利用钙指示剂Fluo-4,采用流式细胞术检测健康对照和脓毒症患者中性粒细胞内钙水平,以及钙螯合剂BAPTA-AM与EDTA共同孵育的健康对照中性粒细胞内的钙水平。结果与健康对照组相比,脓毒症患者中性粒细胞内钙水平显著下降(P<0.01)。钙螯合剂BAPTA-AM与EDTA联合处理后,健康对照中性粒细胞内钙水平显著下降(P<0.01)。微流控技术发现细胞内钙下降后,中性粒细胞在微流控中的迁移速度、迁移距离以及间隙通过率均显著降低(P<0.01)。结论脓毒症患者中性粒细胞内钙水平降低可能是导致中性粒细胞迁移能力下降的重要原因。本研究为通过调节细胞内钙水平改善中性粒细胞迁移障碍提供了新的研究思路。Objective To investigate the relationship between intracellular calcium level and neutrophil migration dysfunction in patients with sepsis.Methods This study retrospectively collected 21 blood samples of patients with sepsis in the First Hospital of Jilin University from December 2017 to September 2018,and 20 healthy people were included as the control group.Peripheral blood neutrophils were isolated from the healthy controls and patients with sepsis using magnetic-activated cell sorting.Multichannel microfluidic microarray technology was used to detect the chemotactic migration of neutrophils.The levels of calcium in neutrophils from healthy controls and sepsis patients as well as in neutrophils from healthy controls that were pretreated with calcium chelators BAPTA-AM and EDTA were detected by flow cytometry using the calcium indicator Fluo-4.Results The intracellular calcium levels were lower in neutrophils from sepsis patients than in those from healthy controls(P<0.01).BAPTA-AM and EDTA could reduce the calcium level in neutrophils of healthy controls(P<0.01).Microfluidics revealed that the migration speed,distance and gap-passing rate of neutrophils in microfluidics were significantly reduced after the decrease of intracellular calcium(P<0.01).Conclusions Reduced calcium levels in neutrophils of patients with sepsis may be closely related to the decreased cell migration.This study suggests that the migration impairment of neutrophils can be improved by regulating intracellular calcium levels,which provides a new idea for further research.

关 键 词：中性粒细胞 脓毒症 细胞迁移 细胞内钙水平 微流控技术 

分 类 号：R459.7[医药卫生—急诊医学]