脓毒症患者支气管肺泡灌洗液中肺泡巨噬细胞表型分析

张尧尧; 吴强; 高山; 侯婷婷; 汪华学; 章凤; 张永

doi:10.16766/j.cnki.issn.1674-4152.001937

脓毒症患者支气管肺泡灌洗液中肺泡巨噬细胞表型分析

doi: 10.16766/j.cnki.issn.1674-4152.001937

张尧尧¹,
吴强²,
高山¹,
侯婷婷¹,
汪华学²,
章凤¹,
张永^1, ,

1.
蚌埠医学院第一附属医院呼吸与危重症医学科，安徽蚌埠 233004
2.
蚌埠医学院第一附属医院重症医学科，安徽蚌埠 233004

基金项目:

安徽省重点研究与开发计划项目 1804h08020287

详细信息

通讯作者:
张永，E-mail：zsuzy@126.com

中图分类号: R631 R446
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- 被引次数: 0
出版历程
- 收稿日期: 2020-12-01
- 网络出版日期: 2022-02-16

Alveolar macrophage phenotypes in bronchoalveolar lavage fluid from sepsis

1.
Department of Pulmonary and Critical Care Medicine, the First Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui 233004, China

摘要

摘要: 目的脓毒症是一种死亡率较高的感染性疾病，以免疫紊乱为特征。巨噬细胞作为一种重要的抗原提呈细胞，在先天免疫和获得性免疫中发挥不可或缺的作用, 其接受不同刺激时可分化为不同的表型——M1和M2，分别具有促进炎症、组织修复重塑的功能。本研究拟观察脓毒症患者支气管肺泡灌洗液(BALF)中肺泡巨噬细胞(alveolar macrophage，AM)表型变化，并分析其不同表型与脓毒症患者预后的关系。方法选取2020年7—10月重症医学科(ICU)和呼吸与危重症医学科(PCCM)病房中29例脓毒症患者，分别于第1天和第7天行支气管肺泡灌洗(bronchoalveolar lavage, BAL)。采用CD68、CD86和CD206三种流式抗体检测AM在BALF中的比例。以CD68抗体作为AM的标记，CD86和CD206抗体分别作为M1和M2标记。同时收集基础临床资料，根据28 d病死率将受试者分为存活组(20例)和死亡组(9例)。结果流式细胞术显示M1和M2型AM的形态结构不同，M1的细胞大小和细胞颗粒复杂度均大于M2。存活组第1天BALF中M1和M2分别为(21.39±9.91)%、(4.66±2.53)%，第7天分别为(3.62±3.31)%、(44.41±7.47)%，M1比例较第1天明显下降且M2比例明显上升，差异有统计学意义(均P < 0.05)；死亡组第1天M1和M2分别为(18.37±2.75)%、(20.40±2.27)%，第7天分别为(7.00±1.98)%、(19.69±4.24)%，M1比例较第1天下降(P < 0.05)且M2比例持续高于M1，M2比例第7天与第1天比较差异无统计学意义(P>0.05)。结论早期动态监测肺泡巨噬细胞表型变化，可能有助于预测脓毒症患者28 d内预后情况。
- 脓毒症 /
- 肺泡巨噬细胞 /
- 抗原提呈细胞 /
- 支气管肺泡灌洗液 /
- 流式细胞术
Abstract: Objective Sepsis is an immune disorder and an infectious disease with high mortality. As important antigen-presenting cells, macrophages play an indispensable role in innate immunity and adaptive immunity. Macrophages may polarise into two distinct phenotypes presenting as M1 and M2 when cells accept different stimuli. M1 macrophages and M2 macrophages enhance inflammation and tissue repair and remodelling, respectively. This study aimed to observe changes in alveolar macrophage (AMs) phenotypes in the bronchoalveolar lavage fluid (BALF) of patients with sepsis and analyse the correlation between the two distinct AM phenotypes and prognosis of patients with sepsis. Methods Twenty-nine patients who experienced consecutive sepsis and admitted to the intensive care unit of the Department of Pulmonary and Critical Care Medicine from July 2020 to December 2020 were included in this study. These patients underwent bronchoalveolar lavage on the first and the seventh days after the admission. The proportion of AMs in BALF was assessed via flow cytometry by using three colour fluorescent stains. CD68 was used as the marker of, total AM whereas CD86 and CD206 were used as M1 and M2 AM markers, respectively. The basic clinical data of the patients were collected and analysed. They were divided into a survival group and a non-survivor group according to mortality data within 28 days. Results The morphological structures of the M1 and M2 AMs were different. Flow cytometry revealed that the M1 AMs have a larger cell size and granularity than the M1 AMs. In the survivor group, the proportion of M1 and M2 in BALF were (21.39±9.91)% and (4.66±2.53)%, respectively, on the first day. On the seventh day, the proportion of M1 and M2 in BALF were (3.62±3.31)% and (44.41±7.47)%, respectively. The percentage of M2 and M1 AMs from BALF increased and decreased, respectively, on the seventh day compared with that on the first day (all P < 0.05). In the nonsurvivor group, the proportion of M1 and M2 in BALF were (18.37±2.75)% and (20.40±2.27)%, respectively, on the first day and (7.00±1.98)% and (19.69±4.24)%, respectively, on the seventh day., the percentage of M1 decreased, whereas that of M2 continued to increase compared with that of M1 (all P < 0.05). No statistically significant differences were observed between the first and the seventh days (P>0.05). Conclusion Early dynamic monitoring of changes in AM phenotypes can help predict the prognosis of patients with sepsis within 28 days.
- Sepsis /
- Alveolar macrophages /
- Antigen presenting cell /
- Bronchoalveolar lavage fluid /
- Flow cytometry

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图 1 Flowjo软件绘制的肺泡巨噬细胞亚型图

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图 2 流式细胞仪检测结果

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表 1 存活组与死亡组脓毒症患者M1和M2型AM比例比较(x±s, %)

组别	例数	M1		M2
组别	例数	第1天	第7天	第1天	第7天
存活组	20	21.39±9.91	3.62±3.31^a	4.66±2.53	44.41±7.47^a
死亡组	9	18.37±2.75	7.00±1.98^a	20.40±2.27	19.69±4.24
t值		1.261	-2.825	-15.966	9.222
P值		0.219	0.009	< 0.001	< 0.001
注：与组内第1天比较，^aP < 0.05。

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