冷诱导RNA结合蛋白在大鼠低体温性急性肺损伤中的表达变化

徐耀迪; 王佳新; 陈旭昕; 张春阳; 王凡; 丁毅伟; 彭聪; 肖漓; 韩志海

doi:10.3969/j.issn.2095-5227.2023.04.011

冷诱导RNA结合蛋白在大鼠低体温性急性肺损伤中的表达变化

doi: 10.3969/j.issn.2095-5227.2023.04.011

徐耀迪^{1, 2,},
王佳新^{2, 3},
陈旭昕²,
张春阳²,
王凡²,
丁毅伟²,
彭聪⁴,
肖漓⁴,
韩志海^{2, 1, ,}

1.
华南理工大学医学院，广东广州　510006
2.
解放军总医院第六医学中心呼吸与危重症医学科，北京　100048
3.
解放军总医院研究生院，北京　100853
4.
解放军总医院第八医学中心呼吸与危重医学部研究所，北京市器官移植与免疫调节重点实验室，北京　100091

基金项目: 军队后勤科研项目(CLB20J019)

详细信息

作者简介:
徐耀迪，女，硕士，医师。研究方向：急性肺损伤。Email: xu_yaodi@163.com

通讯作者:
韩志海，男，主任医师，博士生导师。Email: hanzhihai@301hospital.com.cn

中图分类号: R563
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- 被引次数: 0
出版历程
- 收稿日期: 2022-09-21
- 网络出版日期: 2023-04-18
- 刊出日期: 2023-04-28

Expression of cold-induced RNA-binding protein in hypothermic acute lung injury model in rats

XU Yaodi^{1, 2
,},
WANG Jiaxin^{2, 3},
CHEN Xuxin²,
ZHANG Chunyang²,
WANG Fan²,
DING Yiwei²,
PENG Cong⁴,
XIAO Li⁴,
HAN Zhihai^{2, 1
, ,}

1.
School of Medicine, South China University of Technology, Guangzhou 510006, Guangdong Province, China
2.
Department of Respiratory and Critical Care Medicine, the Sixth Medical Center, Chinese PLA General Hospital, Beijing 100048, China
3.
Graduate School, Chinese PLA General Hospital, Beijing 100853, China
4.
Institute of Respiratory and Critical Care Medicine, the Eighth Medical Center, Chinese PLA General Hospital, Beijing Key Laboratory of Organ Transplantation and Immunology Regulatory, Beijing 100091, China

More Information

Corresponding author: HAN Zhihai. Email: hanzhihai@301hospital.com.cn

摘要

摘要: 背景海上遇难者落水后常死于海水长时间浸泡后的低体温及相关并发症，肺是低体温损伤的重要器官之一，但目前针对低体温造成急性肺损伤(acute lung injury，ALI)的研究相对匮乏。目的探讨冷诱导RNA结合蛋白(cold-inducible RNA-binding protein，CIRP)在低体温性ALI大鼠模型中的变化及其可能的作用机制。方法将40只雄性成年SD大鼠随机分为0 h组(0 h，8只)和实验组(32只)。实验组分别用低温海水浸泡12 h、16 h、20 h、24 h(每组8只)，构建低体温性ALI动物模型。ELISA检测白细胞介素-6(interleukin-6，IL-6)、IL-1β、细胞外CIRP的含量，对肺组织进行HE染色、TUNEL染色和CIRP免疫组织化学染色，qRT-PCR检测CIRP mRNA的表达。结果与0 h组相比，实验组出现不同程度肺损伤，病理切片可见肺泡壁增厚或结构破坏，肺泡腔内出血，伴中性粒细胞和淋巴细胞浸润等； 16 h、20 h、24 h实验组血清中IL-6的表达显著增加，20 h、24 h实验组血清中IL-1β表达显著增加，差异均有统计学意义(P＜0.01)；各实验组大鼠血清中CIRP表达增加均有统计学差异(12 h、16 h、20 h组，P＜0.05；24 h组，P＜0.01)。TUNEL结果显示，各实验组的大鼠肺组织出现不同程度的细胞凋亡现象(P＜0.01)；通过炎症因子的表达水平、肺组织病理改变以及细胞凋亡情况选择低温海水浸泡24 h为最佳构建模型时间，免疫组织化学染色和qRT-PCR观察到24 h组肺组织内CIRP的表达增加。结论肺组织和外周血清的CIRP在低体温性ALI大鼠模型中随时间延长而表达增高，提示其可能与低体温致ALI相关，同时与炎症因子IL-1β、IL-6水平呈现一致性趋势变化，因此推测CIRP可能通过促进炎症因子释放发挥作用，本研究初步证实了CIRP在肺组织区域和整体与低体温ALI的相关性。
- 低体温 /
- 肺损伤 /
- 大鼠 /
- 冷诱导RNA结合蛋白 /
- 炎症因子
Abstract: Background Marine casualties often die from hypothermia and related complications after prolonged immersion in seawater, and lungs are one of the important organs vulnerable to hypothermia injury, but there is a relative lack of researches on acute lung injury (ALI) caused by hypothermia. Objective To investigate the changes of cold-induced RNA-binding protein (CIRP) in rats with hypothermic ALI and its possible mechanism. Methods An animal model of hypothermic ALI was constructed, and 40 male adult SD rats were randomly divided into control group (0 h, n=8) and experimental group (n=32). The experimental groups were immersed in low-temperature seawater for 12 h, 16 h, 20 h, and 24 h (n=8). The contents of IL-6, IL-1β and CIRP were detected by ELISA. HE staining, TUNEL staining and CIRP immunohistochemical staining were performed on lung tissue. The expression of CIRP mRNA was detected by qRTPCR. Results Compared with the control group, the experimental groups had different degrees of lung injury, and pathological sections showed thickening or structural damage of the alveolar wall, hemorrhage in the alveolar cavity, and infiltration of neutrophils and lymphocytes. Compared with the control group, the expression of IL-6 in the serum of the 16 h, 20 h and 24 h experimental groups increased, and IL-1β in the serum of the 20 h and 24 h experimental groups also increased, the differences were statistically significant (all P<0.01). Compared with the control group, the expression of CIRP in serum of rats in each experimental group increased significantly (P<0.05 in 12 h, 16 h, 20 h groups, P<0.01 in 24 h group); TUNEL results showed that there were different degrees of apoptosis in the lung tissue of rats in each experimental group (P<0.01). According to the expression level of inflammatory factors, low temperature seawater immersion for 24 hours was selected as the best time to establish the model. Immunohistochemical staining and qRT PCR showed that the expression of CIRP in lung tissue increased in the 24 hours group. Conclusion The expression of CIRP in lung tissue and peripheral serum increases with time in the rat model of hypothermic ALI, suggesting that CIRP is closely related to hypothermic ALI. And CIRP is consistent with the levels of inflammatory cytokines IL-1 β and IL-6, so it is speculated that CIRP may play a role in promoting the release of inflammatory cytokines. This study preliminarily confirms the correlation between CIRP in lung tissue region and overall with hypothermic ALI.
- hypothermia /
- lung injury /
- rat /
- cold-inducible RNA-binding protein /
- inflammatory cytokines

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图 1 各组大鼠肺组织病理表现

Figure 1. Pathological manifestations of lung tissues of rats in each group

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图 2 各组大鼠肺组织损伤评分和肺系数比较

Figure 2. Comparison in rat lung tissue damage score and lung coefficient between different groups

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图 3 各组大鼠肺组织凋亡情况比较

Figure 3. Apoptosis of lung tissue of rats

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图 4 各组大鼠肺组织凋亡阳性率比较

Figure 4. Comparison in positive rate of apoptosis in lung tissue of rats

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图 5 血清中IL-6 (A)、IL-1β (B)、eCIRP (C)的浓度

Figure 5. Concentration of IL-6 (A), IL-1β (B) and eCIRP (C) in serum

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图 6 eCIRP和IL-6、IL-1β的相关性分析

Figure 6. Correlation of eCIRP with IL-6 and IL-1β

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图 7 肺组织中CIRP mRNA的表达

Figure 7. Expression levels of CIRP mRNA in lung tissue

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图 8 大鼠肺组织中iCIRP的表达(免疫组化，200×)

Figure 8. Expression level of iCIRP in rat lung tissue (IHC, 200×)

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