肺炎支原体社区获得性呼吸窘迫综合征毒素对单核细胞亚群的影响

doi:10.11958/20230118

天津医药 ›› 2023, Vol. 51 ›› Issue (10): 1080-1083.doi: 10.11958/20230118

肺炎支原体社区获得性呼吸窘迫综合征毒素对单核细胞亚群的影响

张岑(), 王志华^△(), 杨磊

天津市南开医院重症医学科（邮编300100）

收稿日期:2023-02-02 修回日期:2023-03-09 出版日期:2023-10-15 发布日期:2023-10-18
通讯作者: ^∆E-mail：wzc927@126.com
作者简介:张岑（1984），女，硕士，主要从事重症医学及呼吸危重症方面研究。E-mail：zhangcen126@126.com
基金资助:
天津市卫生健康委员会中西医结合科研课题(2021094)

Effects of Mycoplasma pneumoniae community acquired respiratory distress syndrome toxin on monocyte subsets

ZHANG Cen(), WANG Zhihua^△(), YANG Lei

Department of Critical Care, Tianjin Nankai Hospital, Tianjin 300100, China

Received:2023-02-02 Revised:2023-03-09 Published:2023-10-15 Online:2023-10-18
Contact: ^∆E-mail：wzc927@126.com

张岑, 王志华, 杨磊. 肺炎支原体社区获得性呼吸窘迫综合征毒素对单核细胞亚群的影响[J]. 天津医药, 2023, 51(10): 1080-1083.

ZHANG Cen, WANG Zhihua, YANG Lei. Effects of Mycoplasma pneumoniae community acquired respiratory distress syndrome toxin on monocyte subsets[J]. Tianjin Medical Journal, 2023, 51(10): 1080-1083.

摘要/Abstract

摘要：

目的探讨肺炎支原体社区获得性呼吸窘迫综合征（CARDS）毒素对小鼠外周血单核细胞亚群的影响。方法制备重组CARDS毒素。32只小鼠按随机数字表分为模型组和正常对照组，每组16只。模型组每只小鼠气管注射40 μL含有50 μg CARDS毒素的PBS，正常对照组气管注射同等剂量的PBS。造模后24 h腹主动脉采血，流式细胞术检测外周血单核细胞亚群（CD11b⁺Ly6G^-Ly6C⁺）比例变化；采用HE染色观察肺组织病理学变化，进行肺组织损伤评分；Real-time PCR检测肺组织中白细胞介素（IL）-1β和IL-18表达水平；采用酶联免疫吸附试验检测外周血IL-1β和IL-18水平。结果与正常对照组相比，模型组小鼠单核细胞CD11b⁺Ly6G^-Ly6C⁺比例升高（P<0.05）；HE染色可见模型组肺泡壁增厚、炎性细胞浸润增加，病理评分显著增高（P<0.01）；肺组织及血清炎性因子IL-1β和IL-18 表达水平均增高（P<0.01）。结论 CARDS毒素诱导的小鼠中Ly6C⁺单核细胞占比升高，其可能与肺炎支原体肺炎的发病密切相关。

关键词: 肺炎, 支原体, 毒素类, 生物学, 单核细胞, 白细胞介素18, 社区获得性呼吸窘迫综合征

Abstract:

Objective To investigate the effect of Mycoplasma pneumoniae community-acquired respiratory distress syndrome (CARDS) toxin on peripheral blood mononuclear cell subsets in mice. Methods The recombinant CARDS toxin was prepared. Thirty-two mice were divided into the model group and the normal control group according to random number table with 16 mice in each group. Each mouse in the model group was injected with 40 μL PBS containing 50 μg of CARDS toxin. In the normal control group, the same dose of PBS was injected into trachea. Blood samples were collected from abdominal aorta 24 h after the modeling, and the proportion of peripheral monocyte subsets (CD11b⁺Ly6G^-Ly6C⁺) was detected by flow cytometry. The mice were sacrificed, and lung histopathological changes were observed by HE staining, and the lung tissue injury score was performed. The expression levels of interleukin (IL)-1β and IL-18 in lung tissue were detected by Real-time PCR. The levels of IL-1β and IL-18 in peripheral blood were determined by enzyme-linked immunosorbent assay. Results Compared with the normal control group, CD11b⁺Ly6G^-Ly6C⁺ratio of mouse monocytes was increased in the model group (P<0.05). HE staining showed that alveolar wall thickened, inflammatory cell infiltration increased, pathological score increased significantly (P<0.01), the expression levels of IL-1β and IL-18 mRNA in lung tissue and contents of inflammatory cytokines IL-1β and IL-18 in serum increased significantly (P<0.01). Conclusion The increased proportion of Ly6C⁺ monocytes in mice induced by CARDS toxin may be closely related to the incidence of Mycoplasma pneumoniae pneumonia.

Key words: pneumonia, Mycoplasma, toxins, biological, monocytes, interleukin-18, community-acquired respiratory distress syndrome

中图分类号:

R563.1

图/表 4

表1 PCR引物序列

Tab.1 The primer sequence of PCR

基因名称	引物序列（5′→3′）
GAPDH	上游：AGGTCGGTGTGAACGGATTTG 下游：GGGGTCGTTGATGGCAACA	202
IL-1β	上游：GAAATGCCACCTTTTGACAGTG 下游：TGGATGCTCTCATCAGGACAG	116
IL-18	上游：GTGAACCCCAGACCAGACTG 下游：CCTGGAACACGTTTCTGAAAGA	202

图1 小鼠外周血CD11b+Ly6G-Ly6C+单核细胞亚群的比例

Fig.1 Proportion of CD11b+Ly6G-Ly6C+ monocyte subsets in peripheral blood of mice

图2 肺组织病理学表现（HE染色，×200）

Fig.2 Pathological findings of lung tissue (HE staining, ×200)

表2 2组肺组织和血清IL-1β、IL-18表达水平比较 (n=3,$\bar{x}±s$)

Tab.2 Comparison of mRNA expression levels of IL-1β and IL-18 in lung tissue and serum between two groups

组别	肺组织		血清/（ng/L）
组别	IL-1β	IL-18	IL-1β	IL-18
正常对照组	1.00±0.02	1.00±0.38	72.49±9.34	95.96±17.79
模型组	13.65±1.17	13.23±2.18	214.39±21.73	333.61±39.21
t	18.670^＊＊	9.735^＊＊	13.417^＊＊	12.341^＊＊

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肺炎支原体社区获得性呼吸窘迫综合征毒素对单核细胞亚群的影响

Effects of Mycoplasma pneumoniae community acquired respiratory distress syndrome toxin on monocyte subsets

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