基于 CRISPR/Cas9 技术建立IL-10rb基因敲除过敏性哮喘小鼠模型

doi:10.3969/j.issn.1006-7795.2026.04.012

首都医科大学学报 ›› 2026, Vol. 47 ›› Issue (4): 715-721.doi: 10.3969/j.issn.1006-7795.2026.04.012

基于 CRISPR/Cas9 技术建立IL-10rb基因敲除过敏性哮喘小鼠模型

张旭^1,2,李景云¹,谭娟²,于渤洋¹,张媛²,张罗^1，2,3*

1.北京市耳鼻咽喉科研究所，过敏性疾病北京实验室（北京市教育委员会），慢性鼻病新药及诊断技术研发北京市重点实验室，北京 100005；2.首都医科大学附属北京同仁医院变态反应（鼻过敏）科，北京 100730；3.首都医科大学附属北京同仁医院耳鼻咽喉头颈外科，北京 100730

收稿日期:2026-04-08 修回日期:2026-05-12 出版日期:2026-08-21 发布日期:2026-07-26
通讯作者: 张罗 E-mail:dr.luozhang@139.com
基金资助:
国家自然科学基金青年科学基金项目（81800886），北京市自然科学基金青年科学基金项目（7164247）。

Construction of the IL-10rb genes mouse model with allergic asthma disease based on CRISPR/Cas9 technology

Zhang Xu^1,2， Li Jingyun¹， Tan Juan²， Yu Boyang¹， Zhang Yuan²，Zhang Luo^1，2,3*

1.Beijing Laboratory of Allergic Diseases, Beijing Municipal Education Commission and Beijing Key Laboratory of New Medicine and Diagnostic Technology Research for Nasal Disease, Beijing Institute of Otolaryngology, Beijing 100005, China；2.Department of Allergy, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China; 3.Department of Otolaryngology Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China

Received:2026-04-08 Revised:2026-05-12 Online:2026-08-21 Published:2026-07-26
Supported by:
This study was supported by the Young Scientists Fund of the National Natural Science Foundation of China (81800886), Natural Science Foundation of Beijing (7164247).

摘要/Abstract

摘要： 目的利用CRISPR/Cas9基因编辑技术构建IL-10rb基因敲除小鼠（IL-10rb^－/－），并评估该基因敲除对卵清蛋白（ovalbumin,OVA）诱导的过敏性哮喘小鼠模型病理表现的影响。方法针对IL-10rb基因外显子1－2区域设计靶向gRNA，将CRISPR/Cas9体系显微注射于C57BL/6J小鼠的受精卵内。仔鼠出生后7 d经聚合酶链式反应（polymerase chain reaction，PCR）鉴定筛选出IL-10rb^－/－基因型的F0代阳性小鼠。将小鼠分为 4 组：野生型（wild-type,WT）组、 IL-10rb^－/－组、WT-OVA 哮喘组、 IL-10rb^－/－-OVA哮喘组。通过肺功能检测以及肺组织苏木精－伊红（hematoxylin-eosin，H&E）、过碘酸-希夫（periodic acid Schiff，PAS）和Masson染色，评估小鼠气道炎症浸润、杯状细胞化生及胶原沉积情况。结果 PCR鉴定证实成功构建稳定遗传的 IL-10rb纯合敲除小鼠品系。肺功能检测显示，与WT组相比，IL-10rb^－/－组、WT-OVA哮喘组及IL-10rb^－/－-OVA 哮喘组小鼠呼吸系统顺应性均显著降低（P<0.05）；与WT-OVA组相比，IL-10rb^－/－-OVA组小鼠呼吸系统顺应性进一步降低（P<0.05）。主气道阻力方面，与WT组相比，WT-OVA组及IL-10rb^－/－-OVA组小鼠气道阻力及组织弹性均显著升高（P<0.05）。肺组织病理结果显示，与WT组相比，IL-10rb^－/－组、WT-OVA组和IL-10rb^－/－-OVA组小鼠炎症细胞浸润显著增多；IL-10rb^－/－-OVA组气道杯状细胞化生较WT-OVA组更为显著；与WT组和IL-10rb^－/－组相比，OVA致敏组小鼠（WT-OVA组和IL-10rb^－/－-OVA组）胶原蛋白沉积明显增强。结论基于CRISPR/ Cas9基因编辑技术可成功构建IL-10rb基因敲除小鼠，该小鼠能够呈现过敏性哮喘的典型病理特征，可作为深入研究过敏性哮喘发病机制及开发靶向干预策略的动物模型。

关键词: CRISPR/Cas9, IL-10rb, 基因敲除, 过敏性哮喘, 气道炎症, 动物模型

Abstract: Objective To generate IL-10rb gene knockout mice (IL-10rb^－/－) using CRISPR/Cas9 gene editing technology and to evaluate the impact of this knockout on the pathological manifestations of an ovalbumin (OVA)-induced allergic asthma mouse model.Methods A targeting gRNA was designed against exons 1-2 of the IL-10rb gene. The CRISPR/Cas9 system was microinjected into fertilized eggs of C57BL/6J mice. F0 positive mice with the IL-10rb^－/－ genotype were identified and selected by polymerase chain reaction (PCR) at 7 day after birth. The mice were divided into four groups: wild-type (WT) group, IL-10rb^－/－ group, WT-OVA asthma group, and IL-10rb^－/－-OVA asthma group. Lung function testing as well as hematoxylin-eosin(H&E), periodic acid Schiff(PAS), and Masson staining of lung tissues were performed to assess airway inflammatory infiltration, goblet cell metaplasia, and collagen deposition.Results PCR identification confirmed the successful generation of a stably heritable IL-10rb homozygous knockout mouse strain. Lung function tests showed that compared with the WT group, the IL-10rb^－/－ group, WT-OVA asthma group, and IL-10rb^－/－-OVA asthma group all exhibited significantly decreased respiratory system compliance (P<0.05). Compared with the WT-OVA group, the IL-10rb^－/－-OVA group showed a further decrease in respiratory system compliance (P<0.05). In terms of main airway resistance, compared with the WT group, airway resistance and tissue elasticity in the WT-OVA group and the IL-10rb^－/－-OVA group were significantly increased (P<0.05). Lung histopathology revealed that compared with the WT group, the inflammatory cell infiltration in the IL-10rb^－/－ group, WT-OVA group, and IL-10rb^－/－-OVA group was significantly increased. Airway goblet cell metaplasia was more pronounced in the IL-10rb^－/－-OVA group than that in the WT-OVA group. Collagen deposition was markedly enhanced in the OVA-sensitized groups (WT-OVA and IL-10rb^－/－-OVA) compared with the WT and IL-10rb^－/－ groups.Conclusion IL-10rb gene knockout mice can be successfully generated using CRISPR/Cas9 gene editing technology. These mice can recapitulate the typical pathological features of allergic asthma and serve as an animal model for further investigating the pathogenesis of allergic asthma and developing targeted intervention strategies.

Key words: CRISPR/Cas9, IL-10rb, gene knockout, allergic asthma, airway inflammation, animal model

中图分类号:

R562.2+5

张旭, 李景云, 谭娟, 于渤洋, 张媛, 张罗. 基于 CRISPR/Cas9 技术建立IL-10rb基因敲除过敏性哮喘小鼠模型[J]. 首都医科大学学报, 2026, 47(4): 715-721.

Zhang Xu, Li Jingyun, Tan Juan, Yu Boyang, Zhang Yuan, Zhang Luo. Construction of the IL-10rb genes mouse model with allergic asthma disease based on CRISPR/Cas9 technology[J]. Journal of Capital Medical University, 2026, 47(4): 715-721.

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基于 CRISPR/Cas9 技术建立IL-10rb基因敲除过敏性哮喘小鼠模型

Construction of the IL-10rb genes mouse model with allergic asthma disease based on CRISPR/Cas9 technology

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