程序性死亡[蛋白]-1抑制剂诱导小鼠心肌炎模型的建立

陈怡帆; 程蕾蕾; 沈毅辉; 张卉; 汪雪君; 许宇辰; 葛均波

doi:10.19401/j.cnki.1007-3639.2022.07.004

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程序性死亡[蛋白]-1抑制剂诱导小鼠心肌炎模型的建立

论著 | 更新时间：2025-12-31

- 程序性死亡[蛋白]-1抑制剂诱导小鼠心肌炎模型的建立
- Establishment of a mouse myocarditis model induced by programmed death-1 inhibitor
- 中国癌症杂志 2022年32卷第7期页码：606-613
- 作者机构：
  
  1. 复旦大学附属中山医院心内科,上海 200032
  2. 上海市心血管病研究所,上海 200032
  3. 国家放射与治疗临床医学研究中心,上海 200032
  4. 复旦大学附属中山医院心脏超声诊断科,上海 200032
- 作者简介：
  
  [ "陈怡帆（ORCID: 0000-0001-9161-8268）,博士,E-mail: 21111210020@m.fudan.edu.cn。" ]
  程蕾蕾（ORCID: 0000-0003-0677-7892）,博士,主任医师,E-mail: cheng.leilei@zs-hospital.sh.cn。
- 基金信息：
  
  国家自然科学基金(82170359);上海介入治疗工程技术研究中心(19DZ2250300);放射与治疗临床医学研究中心项目(19MC1910300)
- DOI：10.19401/j.cnki.1007-3639.2022.07.004
  中图分类号： R542.2+1
- 收稿：2022-03-08，
  
  纸质出版：2022-07-30
- 稿件说明：
移动端阅览
陈怡帆, 程蕾蕾, 沈毅辉, 等. 程序性死亡[蛋白]-1抑制剂诱导小鼠心肌炎模型的建立[J]. 中国癌症杂志, 2022,32(7):606-613.

Yifan CHEN, Leilei CHENG, Yihui SHEN, et al. Establishment of a mouse myocarditis model induced by programmed death-1 inhibitor[J]. China Oncology, 2022, 32(7): 606-613.
陈怡帆, 程蕾蕾, 沈毅辉, 等. 程序性死亡[蛋白]-1抑制剂诱导小鼠心肌炎模型的建立[J]. 中国癌症杂志, 2022,32(7):606-613. DOI： 10.19401/j.cnki.1007-3639.2022.07.004.

Yifan CHEN, Leilei CHENG, Yihui SHEN, et al. Establishment of a mouse myocarditis model induced by programmed death-1 inhibitor[J]. China Oncology, 2022, 32(7): 606-613. DOI： 10.19401/j.cnki.1007-3639.2022.07.004.

摘要

背景与目的：

程序性死亡[蛋白

]

-1（programmed death-1

PD-1）抑制剂诱发的心肌炎亟待通过动物模型探究治疗靶点。本研究旨在探索此类自身免疫性心肌炎小鼠模型的最佳造模方法。

方法：

选取6周龄健康雄性BALB/c小鼠30只

分别编号并随机分为对照组（control组）、自身免疫性心肌炎组（Tn

#x02160;组）和免疫检查点抑制剂（immune checkpoint inhi

bitor

ICI）相关心肌炎组（Tn

#x02160;+anti-PD-1组）

每组10只。除control组外

分别于第1、7天给小鼠皮下注射0.1 mL含有0.25 mg小鼠心肌Tn

#x02160;肽段的完全弗氏佐剂。Tn

#x02160;+anti-PD-1组自第7天起

每次5 mg/kg腹腔注射PD-1抑制剂

每2 d给药1次

共5次

累积剂量25 mg/kg。观察小鼠的一般状态、死亡率、心脏指数、超声心动图、心肌病理学改变、血清肌酸激酶（creatine kinase

CK）及CK同工酶（CK isoenzyme

CK-MB）水平。

结果：

与control组相比

造模第21、56天

#x02160;和Tn

#x02160;+anti-PD-1组体重均显著下降（

0.05

0.01）

两组间体重有显著差异（

0.05）

进食量均显著下降（

0.05

0.01）。两组的死亡率在第21天分别为0%和10%

在第56天分别为10%和20%。第56天

#x02160;组心脏指数未显著增加（

0.05）

#x02160;+anti-PD-1组显著增加（

0.05）

且左心室射血分数（ejection fraction

EF）显著下降（

0.001）。急性心肌炎期

#x02160;组心外膜下见少量炎症细胞浸润

#x02160;+anti-PD-1组心外膜下见大量炎症细胞浸润

心肌细胞坏死。扩张型心肌病期

#x02160;组炎症细胞浸润减少

心肌细胞轻度边界不清、空泡化

#x02160;+anti-PD-1组细胞坏死、空泡化、核异形更显著。第56天

#x02160;+anti-PD-1组血清CK、CK-MB显著升高（

0.001）

且较Tn

#x02160;组升高更明显（CK：

0.01;CK-MB：

0.05）。

结论：

建立了PD-1抑制剂诱导的、低死亡率的小鼠心肌炎模型

以心肌急慢性自身免疫炎性改变、左心室EF下降、心肌酶谱升高为突出特征。设计合成了特殊的小鼠心肌Tn

#x02160;肽段

强化了PD-1抑制剂的造模效果。

Abstract

Background and purpose:

Myocarditis induced by programmed death-1 (PD-1) inhibitors is in urgent need of animal models to explore therapeutic targets. The study aimed to figure out the best modeling method of this typical autoimmune myocarditis in mice.

Methods:

Thirty 6-week-old healthy male BALB/c mice were numbered and randomly divided into control group

autoimmune myocarditis group (Tn

#x02160;group) and immune checkpoint inhibitor (ICI)-related myocarditis group (Tn

#x02160;+anti-PD-1 group)

10 in each group. Except for the control group

mice were subcutaneously injected with 0.1 mL complete Freund

#x02019;s adjuvant containing 0.25 mg of mouse cardiac Tn

#x02160; peptide on day

1 and day 7

respectively. From day 7

#x02160;+anti-PD-1 group received intraperitoneal injection of PD-1 inhibitor at 5 mg/kg each time

once every 2 d

for a total of 5 times with a cumulative dose of 25 mg/kg. The general state

mortality

cardiac index

echocardiography

myocardial pathology and the levels of creatine kinase (CK) and CK isoenzyme (CK-MB) in serum were observed.

Results:

Compared with control group

the mass of mice in both Tn

#x02160; and Tn

#x02160;+anti-PD-1 groups decreased significantly on day 21 and day 56 (

0.05

0.01)

and there was a significant difference between these 2 groups (

0.05). Both had significantly decreased food intake (compared with control group

0.05

0.01). The mortality rates were 0% and 10% on day 21

and 10% and 20% on day 56 in Tn

#x02160; and Tn

#x02160;+anti-PD-1 groups

respectively. On day 56

no significant increase in cardiac index could be observed in Tn

#x02160; group (

0.05)

while a significant rise of cardiac index (

0.05) with a decrease in left ventricular ejection fraction (EF) (

0.001) were detected in Tn

#x02160;+anti-PD-1 group. During the acute myocarditis stage

mild subepicardial inflammatory infiltration was found in Tn

#x02160; group; Severe subepicardial inflammatory infiltration and myocardial cell necrosis were seen in Tn

#x02160;+anti-PD-1 group. During the dilated cardiomyopathy stage

the infiltrated inflammatory cells in Tn

#x02160; group decreased

mild boundaries unclear and cytoplasm vacuolization could be observed; Tn

#x02160;+anti-PD-1 group also had decreased inflammatory infiltration while underwent more severe cell necrosis and vacuolization with nuclear atypia. On day 56

serum CK and CK-MB in Tn

#x02160;+anti-PD-1 group rose significantly (

0.001)

which was more obvious

compared with Tn

#x02160; group (CK:

0.01; CK-MB:

0.05).

Conclusion:

A PD-1 inhibitor-induced myocarditis model with low mortality was established in mice

characterized by acute and chronic autoimmune myocardial inflammation

decreased ejection fraction and increased myocardial enzyme spectrum. A mouse cardiac Tn

#x02160; peptide fragment was particularly designed and synthesized for modeling.

关键词

Keywords

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