不同延迟降温时间对劳力型热射病大鼠病理损伤的影响

doi:10.12122/j.issn.1673-4254.2024.10.03

南方医科大学学报 ›› 2024, Vol. 44 ›› Issue (10): 1858-1865.doi: 10.12122/j.issn.1673-4254.2024.10.03

• • 上一篇

不同延迟降温时间对劳力型热射病大鼠病理损伤的影响

赵金宝¹^,²(), 贾亿卿¹(), 毛汉丁¹, 王世娇³, 徐凡², 李鑫⁴, 陶冶⁵, 薛蕾⁶, 刘树元¹(), 宋青², 周必业¹()

^1.解放军总医院第六医学中心急诊医学科，北京 100048
^2.解放军医学院研究生院，北京 100853
^3.解放军总医院第二医学中心内分泌科，北京 100853
^4.解放军总医院第三医学中心急诊医学科，北京 100039
^5.解放军总医院京中医疗区黄寺门诊部，北京 100120
^6.北京卫戍区丰台第一退休干部休养所，北京 100161

收稿日期:2024-04-02 出版日期:2024-10-20 发布日期:2024-10-31
通讯作者: 刘树元,周必业 E-mail:zjb130500@163.com;903915664@qq.com;liusydoc@163.com;zhoubiye@qq.com
作者简介:赵金宝，硕士，主治医师，E-mail: zjb130500@163.com
贾亿卿，在读博士研究生，主治医师，E-mail: 903915664@qq.com
第一联系人：赵金宝、贾亿卿共同为第一作者
基金资助:
军事医学创新项目(18CXZ024)

Effect of different delayed cooling time on organ injuries in rat models of exertional heat stroke

Jinbao ZHAO¹^,²(), Yiqin JIA¹(), Handing MAO¹, Shijiao WANG³, Fan XU², Xin LI⁴, Ye TAO⁵, Lei XUE⁶, Shuyuan LIU¹(), Qing SONG², Biye ZHOU¹()

^1.Department of Emergency Medicine, Sixth Medical Center of Chinese PLA General Hospital, Beijing 100048, China
^2.Graduate School of Medical School of Chinese PLA, Beijing 100853, China
^3.Department of Endocrinology, Second Medical Center of Chinese PLA General Hospital, Beijing 100853, China
^4.Department of Emergency Medicine, Third Medical Center of Chinese PLA General Hospital, Beijing 100039, China
^5.Huangsi Clinic, Jingzhong Medical District of Chinese PLA General Hospital, Beijing 100120, China
^6.Beijing Garrison First Cadre Retreat Center, Beijing 100161, China

Received:2024-04-02 Online:2024-10-20 Published:2024-10-31
Contact: Shuyuan LIU, Biye ZHOU E-mail:zjb130500@163.com;903915664@qq.com;liusydoc@163.com;zhoubiye@qq.com

摘要/Abstract

摘要：

目的通过建立劳力型热射病（EHS）大鼠模型，探索不同降温时间对器官病理损伤的影响及可能的机制。方法选取健康成年雄性Wistar大鼠72只建立劳力型热射病动物模型。将大鼠随机分为6组，12只/组：正常对照组（CON）；模型组（EHS）（建模后不进行主动降温）；即刻降温组（建模后立即进行冷水浴降温）；延迟降温A组（建模后延迟5 min进行冷水浴降温）；延迟降温B组（建模后延迟15 min进行冷水浴降温）；延迟降温C组（建模后延迟30 min进行冷水浴降温）。记录各组大鼠核心体温变化，计算降温速率。所有实验大鼠观察24 h后，解剖留取血液样本检测炎症指标白细胞介素-1β、白细胞介素-2、白细胞介素-4、白细胞介素-6、白细胞介素-10、γ-干扰素并进行脏器（肾脏、肺脏、大脑和回肠）病理学检查；若24 h内发生死亡，则立即解剖留取血液样本化验并进行病理取材。结果模型大鼠24 h内死亡例数随着降温治疗的延迟而增加，各组死亡率差异有统计学意义（P<0.05）。降温延迟时间与死亡率呈正相关（r=0.996，P=0.004）。降温速率与死亡率呈负相关（r=-0.961，P=0.009）。炎症因子检测结果显示，各因子在不同降温干预组的浓度变化呈现出较大的异质性。所有模型动物均有明显器官损伤，病理以上皮脱落、水肿、渗出和炎细胞浸润为主；脑组织和肾组织在发病24 h内受损最明显。结论 EHS大鼠模型肾脏、肺脏、大脑和小肠存在显著的非特异性病理损伤，但受损程度并不一致；随着降温的延迟，病理损伤逐渐加重。炎症因子在不同降温时间干预的血浆浓度变化存在显著异质性。

关键词: 热射病, 劳力型, 动物模型, 降温, 病死率

Abstract:

Methods To investigate how the timing of cooling therapy affects organ injuries in rats with exertional heat stroke (EHS) and explore the possible mechanisms. Methods A total of 60 adult male Wistar rat models of EHS were randomized into model group without active cooling after modeling, immediate cooling group with cold water bath immediately after modeling, delayed cooling groups with cold water bath at 5, 15 and 30 min after modeling, with another 12 mice without EHS as the normal control group. The changes in core body temperature of the mice were recorded and the cooling rate was calculated. After observation for 24 h, the mice were euthanized and blood samples were collected for detection of interleukin-1β (IL-1β), IL-2, IL-4, IL-6, IL-10, and interferon-γ, followed by pathological examination of the vital organs. The rats that died within 24 h were immediately dissected for examination. Results The number of deaths of the model rats within 24 h increased significantly with the time of delay of cooling treatment. The delay of cooling was positively correlated (r=0.996, P=0.004) while the cooling rate negatively correlated with the mortality rate (r=-0.961, P=0.009). The inflammatory cytokine levels presented with different patterns of variations among the cooling intervention groups. All the rat models of EHS had significant organ damages characterized mainly by epithelial shedding, edema, effusion, and inflammatory cell infiltration, and brain and renal injuries reached the peak level at 24 h after EHS. Conclusion EHS causes significant nonspecific pathologies of varying severities in the vital organs of rats, and the injuries worsen progressively with the delay of cooling. There is a significant heterogeneity in changes of serum inflammatory cytokines in rats with different timing of cooling intervention following EHS.

Key words: exertional heat stroke, animal models, cooling interventions, mortality

赵金宝, 贾亿卿, 毛汉丁, 王世娇, 徐凡, 李鑫, 陶冶, 薛蕾, 刘树元, 宋青, 周必业. 不同延迟降温时间对劳力型热射病大鼠病理损伤的影响[J]. 南方医科大学学报, 2024, 44(10): 1858-1865.

Jinbao ZHAO, Yiqin JIA, Handing MAO, Shijiao WANG, Fan XU, Xin LI, Ye TAO, Lei XUE, Shuyuan LIU, Qing SONG, Biye ZHOU. Effect of different delayed cooling time on organ injuries in rat models of exertional heat stroke[J]. Journal of Southern Medical University, 2024, 44(10): 1858-1865.

图/表 10

图1 实验大鼠经直肠测量核心温度

Fig.1 Core temperature of a rat model measured via the rectum.

图2 EHS模型大鼠实验平台

Fig.2 Experimental platform of EHS model rats.

表1 实验动物分组及一般资料比较

Tab.1 Body weight, age, and core temperature （Tc） of the rats before and after modeling in different groups (n=12, Mean±SD)

Group	Weight (g)	Age (week)	Pre-modelling Tc (℃)	Post-modelling Tc (℃)
Control	251.90±7.29	7.0±0.8	37.85±0.81	-
EHS	251.75±6.23	6.9±0.8	37.97±0.85	43.62±0.68
Immediate cooling group	252.77±9.01	7.2±0.5	38.07±0.76	43.22±0.68
Delayed cooling group A	250.45±6.36	6.8±0.7	38.06±0.94	43.32±0.99
Delayed cooling group B	250.29±7.47	7.2±0.8	37.95±0.83	43.52±0.73
Delayed cooling group C	250.68±6.81	7.0±0.9	37.68±0.65	43.16±0.75

表2 降温治疗对模型大鼠24 h死亡率的影响

Tab.2 Effect of cooling treatment on 24-h mortality rate of the rat models of EHS (n=12)

Group	Delay time for cooling (min)	Cooling rate (℃/min)	Number of deaths within 24 h	24-h mortality rate
CON	-	-	0	0
EHS	-	0.08±0.05	7	58.3%
Immediate cooling group	0	0.36±0.11*	1	8.3%*
Delayed cooling group A	5	0.26±0.07*^#	2	16.7%*
Delayed cooling group B	15	0.20±0.05*^#	3	25.0%*
Delayed cooling group C	30	0.16±0.04*^{#^}	5	41.7%^#

表3 降温治疗与模型大鼠24 h死亡率的相关性分析

Tab.3 Correlation analysis of delay time of cooling treatment， cooling rate and 24-h mortality rate of the rat models

Variables Statistical indicators		Cooling rate	Delay time for cooling	Mortality rate
Cooling rate	Pearson correlation coefficient	1	-0.912	-0.961
Cooling rate	P	-	0.088	0.009
Delay time for cooling	Pearson correlation coefficient	-0.912	1	0.996
Delay time for cooling	P	0.088	-	0.004
Mortality rate	Pearson correlation coefficient	-0.961	0.996	1
Mortality rate	P	0.009	0.004	-

图3 各组大鼠肾脏HE染色代表图

Fig.3 Representative images of rat kidney sections in each group (HE staining, original magnification: ×200). A: Control group. B: EHS group. C: Immediate cooling group. D: Delayed cooling group A. E: Delayed cooling group B. F: Delayed cooling group C.

图4 各组大鼠脑组织HE染色代表图

Fig. 4 Representative images of rat brain tissue sections in each group (HE staining, ×200). A: Control group. B: EHS group. C: Immediate cooling group. D: Delayed cooling group A. E: Delayed cooling group B. F: Delayed cooling group C. Black arrows indicate neurons with pyknosis accompanied by collapse and karyolysis of Nissl body.

图5 各组大鼠大肺组织HE染色代表图

Fig. 5 HE staining of rat lung tissue sections in each group (×200). A: Control group. B: EHS group. C: Immediate cooling group. D: Delayed cooling group A. E: Delayed cooling group B. F: Delayed cooling group C.

图6 各组大鼠小肠组织HE染色代表图

Fig. 6 HE staining of rat small intestine tissues in each group (×200). A: Control group. B: EHS group. C: Immediate cooling group. D: Delayed cooling group A. E: Delayed cooling group B. F: Delayed cooling group C.

图7 不同降温组间炎症因子的变化

Fig. 7 Comparison of inflammatory factors among the groups. *P<0.05 vs Control group; #P<0.05 vs EHS group; ^P<0.05 vs immediate cooling group; + P<0.05 vs delayed cooling group A; &P<0.05 vs delayed cooling group B.

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不同延迟降温时间对劳力型热射病大鼠病理损伤的影响

Effect of different delayed cooling time on organ injuries in rat models of exertional heat stroke

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