中华中医药学刊

新型冠状病毒假病毒感染hACE2转基因小鼠的模型建立及评价

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张天宇 ¹ , 吴振起 ²^,³ , 李晓芳 ⁴ , 杨馥语 ¹ , 王语馨 ¹

中华中医药学刊 | 国家项目点击 2026,44(8): 145-149

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中华中医药学刊 |国家项目点击 2026 , 44 (8) : 145 -149

新型冠状病毒假病毒感染hACE2转基因小鼠的模型建立及评价

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张天宇¹, 吴振起²^,³, 李晓芳⁴, 杨馥语¹, 王语馨¹

作者信息

^1.辽宁中医药大学，辽宁　沈阳　110847

^2.辽宁中医药大学附属医院，辽宁　沈阳　110032

^3.沈阳市中医外感病实验室，辽宁　沈阳　110034

^4.辽宁中医药大学附属第二医院，辽宁　沈阳　110034

通讯作者:

吴振起（1974-），男，辽宁庄河人，主任医师，博士研究生导师，博士，研究方向：中医药防治小儿感染性疾病。E-mail：zhenqiwu@163.com。

作者简介:

张天宇（1996-），女，辽宁大连人，助理研究员，博士，研究方向：中医药防治小儿感染性疾病。

Modeling and Evaluation of SARS-CoV-2 Pseudovirus Infection in hACE2 Transgenic Mice

Tianyu ZHANG¹, Zhenqi WU²^,³, Xiaofang LI⁴, Fuyu YANG¹, Yuxin WANG¹

Affiliations

^1.Liaoning University of Traditional Chinese Medicine, Shenyang 110847, Liaoning, China

^2.The Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, Shenyang 110032, Liaoning, China

^3.Shenyang Laboratory of Chinese Medicine and External Diseases, Shenyang 110034, Liaoning, China

^4.The Second Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, Shenyang 110034, Liaoning, China

出版时间: 2026-08-10 doi: 10.13193/j.issn.1673-7717.2026.08.030

文章导航

摘要

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目的

采用含有刺突蛋白（spike，S）质粒、表达荧光素酶（Luciferase）和绿色荧光蛋白（GFP）的新型冠状病毒（severe acute respiratory syndrome coronavirus 2，SARS-CoV-2）假病毒感染K18-hACE2转基因小鼠，探索建立并评价一种可用的动物模型，为后续有效药物靶点筛选提供基础。

方法

（1）将12只K18-hACE2转基因小鼠随机分为对照组和模型组，模型组小鼠通过移液器滴鼻50 μL 1×10⁹PFU/只的病毒液，对照组小鼠予等量空载体病毒液；（2）于造模后第3天观察小鼠Luciferase活体荧光检测后取材；荧光显微镜下观察肺组织GFP荧光强度；光镜下观察小鼠肺组织病理切片；透射电镜观察肺组织超微结构；qPCR法检测肺组织S蛋白表达；ELISA法检测血清中白细胞介素-6（interleukin 6，IL-6）、白细胞介素-10（interleukin 10，IL-10）含量；Western Blot法检测肺组织血管紧张素转换酶2（angiotensin converting enzyme 2，ACE2）蛋白表达。

结果

（1）活体成像可观察到模型组小鼠鼻咽部及双肺有较大荧光面积，肺组织冷冻切片中可见大量绿色荧光，肺组织病理切片及超微结构观察可见病理改变，对照组小鼠肺内未见荧光表达及病理改变；（2）模型组小鼠肺内可见S蛋白mRNA表达（P<0.01）；（3）与对照组小鼠相比，模型组小鼠血清中IL-6表达升高，IL-10表达降低（P<0.01）；（4）与对照组小鼠相比，模型组小鼠肺组织中的ACE2受体含量下降（P<0.01）。

结论

本项研究建立并评价了一种SARS-CoV-2假病毒感染hACE2转基因小鼠的模型，模型的优化仍需后续进一步完善。

关键词

SARS-CoV-2假病毒 / K18-hACE2小鼠 / 动物模型 / 刺突蛋白

Abstract

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Objective

To use severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pseudovirus containing spike protein (S) particles and expressing Luciferase luciferase and GFP fluorescent proteins to infect K18-hACE2 transgenic mice, to explore the establishment and evaluation of a usable animal model, and to provide ideas for the subsequent screening of effective drug targets.

Methods

(1) Twelve K18-hACE2 transgenic mice were randomly divided into control and model groups. The mice in the model group were given 50 μL of 1×10⁹PFU/each viral solution by pipette drops, and those in the control group were given an equal amount of empty vector viral solution. (2) On the 3rd day after modeling, Luciferase in vivo fluorescence was observed and the samples were collected. The lung tissue GFP was observed under fluorescence microscopy. The lung tissue pathology was observed under light microscope. The lung tissue ultrastructure was observed under transmission electron microscope. S protein expression in lung tissue was detected by qPCR. The levels of interleukin 6 (IL-6) and interleukin 10 (IL-10) in serum were detected by ELISA. Western Blot assay was used for angiotensin converting enzyme 2 (ACE2) protein expression in lung tissue.

Results

(1) In vivo imaging revealed a large fluorescence area in the nasopharynx and both lungs of the model mice, a large amount of green fluorescence in frozen sections of lung tissue, and pathological changes in lung histopathological sections and ultramicroscopic findings, while no fluorescence expression or pathological changes were observed in the lungs of the control mice. (2) S protein mRNA expression was seen in the lungs of the model mice (P<0.01). (3) Compared with those of the control mice, the expression of IL-6 in the serum of the model mice was increased and the expression of IL-10 was decreased (P<0.01). IL-6 expression was increased and IL-10 expression was decreased in the serum of mice in the model group compared with those in the control group (P<0.01). (4)　ACE2 receptor content was decreased in lung tissues of the mice in the model group compared to that in the control group (P<0.01).

Conclusion

In this study, a model of SARS-CoV-2 pseudovirus infection in hACE2 transgenic mice was established and evaluated, and the optimization study of the model still needs to be followed up for further improvement.

Key words

SARS-CoV-2 pseudovirus / K18-hACE2 mice / animal model / spike protein

引用本文

张天宇, 吴振起, 李晓芳, 杨馥语, 王语馨. 新型冠状病毒假病毒感染hACE2转基因小鼠的模型建立及评价. 中华中医药学刊, 2026 , 44 (8) : 145 -149 . DOI: 10.13193/j.issn.1673-7717.2026.08.030

Tianyu ZHANG, Zhenqi WU, Xiaofang LI, Fuyu YANG, Yuxin WANG. Modeling and Evaluation of SARS-CoV-2 Pseudovirus Infection in hACE2 Transgenic Mice[J]. Chinese Archives of Traditional Chinese Medicine, 2026 , 44 (8) : 145 -149 . DOI: 10.13193/j.issn.1673-7717.2026.08.030

基金

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国家自然科学基金面上项目(82074494)

参考文献

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2026年第44卷第8期

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文章信息

doi: 10.13193/j.issn.1673-7717.2026.08.030

首发时间：2026-10-20
出版时间：2026-08-10

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基金

国家自然科学基金面上项目(82074494)

作者信息

^1.辽宁中医药大学，辽宁　沈阳　110847

^2.辽宁中医药大学附属医院，辽宁　沈阳　110032

^3.沈阳市中医外感病实验室，辽宁　沈阳　110034

^4.辽宁中医药大学附属第二医院，辽宁　沈阳　110034

通讯作者:

吴振起（1974-），男，辽宁庄河人，主任医师，博士研究生导师，博士，研究方向：中医药防治小儿感染性疾病。E-mail：zhenqiwu@163.com。

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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