Establishment and Evaluation of New Mouse Model of Rheumatoid Arthritis Combined with Interstitial Lung Disease

XU Liting; ZHAO Qingyu; YANG Chao; HE Lianhua; SUN Congcong; GAO Shuangrong; WANG Lili; LIU Chunfang; LIN Na

doi:10.13422/j.cnki.syfjx.20250537

您当前的位置：

首页 >

文章列表页 >

Establishment and Evaluation of New Mouse Model of Rheumatoid Arthritis Combined with Interstitial Lung Disease

更新时间：2026-02-28

- Establishment and Evaluation of New Mouse Model of Rheumatoid Arthritis Combined with Interstitial Lung Disease
  增强出版
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 32, Issue 6, Pages: 81-90(2026)
- 作者机构：
  
  中国中医科学院中药研究所，北京 100700
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20250537
  CLC： R282;R285;R259
- Received：02 March 2025，
  
  Revised：2025-03-15，
  
  Accepted：20 March 2025，
  
  Online First：18 March 2025，
  
  Published：20 March 2026
- 稿件说明：
移动端阅览
许丽婷,赵清玉,杨超等.一种新的类风湿关节炎合并间质性肺病小鼠模型的建立与评价[J].中国实验方剂学杂志,2026,32(06):81-90.

XU Liting,ZHAO Qingyu,YANG Chao,et al.Establishment and Evaluation of New Mouse Model of Rheumatoid Arthritis Combined with Interstitial Lung Disease[J].Chinese Journal of Experimental Traditional Medical Formulae,2026,32(06):81-90.
许丽婷,赵清玉,杨超等.一种新的类风湿关节炎合并间质性肺病小鼠模型的建立与评价[J].中国实验方剂学杂志,2026,32(06):81-90. DOI： 10.13422/j.cnki.syfjx.20250537.

XU Liting,ZHAO Qingyu,YANG Chao,et al.Establishment and Evaluation of New Mouse Model of Rheumatoid Arthritis Combined with Interstitial Lung Disease[J].Chinese Journal of Experimental Traditional Medical Formulae,2026,32(06):81-90. DOI： 10.13422/j.cnki.syfjx.20250537.

摘要

目的

采用牙龈卟啉单胞菌（Pg）感染结合胶原诱导的方法建立类风湿关节炎合并间质性肺病（RA-ILD）小鼠模型，并通过关节、肺和血清等多方面病变特征进行综合评价。

方法

将40只DBA/1小鼠随机分成4组，分别为Control组、Pg感染（Pg）组、胶原诱导性关节炎（CIA）组和Pg感染结合CIA（Pg+CIA）组，每组10只。观察小鼠关节炎临床积分、发病率等关节炎临床症状；微计算机断层扫描（Micro-CT）分析小鼠膝关节病变情况；苏木素-伊红（HE）、马松（Masson）染色检测小鼠膝关节和肺组织病理学形态变化和胶原沉淀情况；免疫组化法检测肺组织

-平滑肌肌动蛋白（

-SMA）、Ⅰ型胶原蛋白（ColⅠ）和纤连蛋白（FN）蛋白表达水平；实时荧光定量聚合酶链式反应（Real-time PCR）检测小鼠肺组织

-SMA、ColⅠ、FN、肿瘤坏死因子-

（TNF-

）、白细胞介素（IL）-6、IL-1

的mRNA表达量；酶联免疫吸附测定法（ELISA）检测小鼠血清中Pg、环瓜氨酸肽（CCP）和免疫球蛋白G（IgG）含量。

结果

关节病变评估：CIA组和Pg+CIA组小鼠关节发病率均为100%，关节红肿、畸形症状明显，受累肢体数分别为27、28分，临床积分分别为68、70分，Pg组关节未观察到明显的临床症状；CIA组和Pg+CIA组关节组织病理学和影像学病变严重，组织病理学评分、骨密度、骨体积分数、骨小梁厚度和骨小梁数量均明显升高，与Control组比较差异显著（

0.01），Pg组未观察到明显的关节炎病理改变。肺部病变评估：Pg+CIA组可见明显的肺泡炎症、间质炎症细胞浸润和肺泡壁增厚等病理表现，胶原纤维蓝染严重，其组织病理学评分和胶原面积比均明显升高，与Control组、Pg组和CIA组比较差异显著（

0.05）；肺中

-SMA、ColⅠ和FN的蛋白和mRNA表达水平明显升高，IL-6、TNF-

和IL-1

mRNA表达水平也明显升高，与Control组比较差异明显（

0.05）。血清学检测结果显示，Pg+CIA组CCP、Pg和IgG含量明显升高，与Control、Pg和CIA 3组比较均差异明显（

0.05）。

结论

Pg感染结合胶原诱导DBA/1小鼠所建立的动物模型，表现出明显的类风湿关节炎和间质性肺病的症状及病理特征，同时伴有高水平的血清CCP，是一种新型的RA-ILD小鼠模型。相关研究将为RA-ILD发病机制探究和新药研发提供实验工具，也为环瓜氨酸肽抗体（ACPA）阳性的RA-ILD动物模型建立提供研究基础。

Abstract

Objective

To establish a mouse model of rheumatoid arthritis with interstitial lung disease （RA-ILD） in DBA/1 mice using

Porphyromonas gingivalis

（Pg） infection combined with collagen-induced

arthritis （CIA）， and to comprehensively evaluate pathological characteristics in joints， lungs， and serum.

Methods

Forty DBA/1 mice were randomly divided into four groups，

i.e

.， Control， Pg infection （Pg）， CIA， and Pg infection combined with CIA （Pg+CIA）， with 10 mice in each group. Arthritis clinical symptoms were evaluated by recording arthritis incidence and clinical scores. Micro-CT scanning was used to assess knee joint pathology. Histopathological changes and collagen deposition in knee joints and lung tissues were analyzed using hematoxylin-eosin （HE） and Masson staining. Immunohistochemistry was performed to detect protein expression of

-smooth muscle actin （

-SMA）， typeⅠ collagen （ColⅠ）， and fibronectin （FN） in lung tissues. Real-time quantitative polymerase chain reaction（Real-time PCR）was used to measure mRNA expression levels of

-SMA， ColⅠ， FN， tumor necrosis factor-

（TNF-

）， interleukin-6 （IL-6）， and IL-1

in lung tissues. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum levels of Pg， cyclic citrullinated peptide （CCP）， and immunoglobulin G （IgG）.

Results

Joint lesions： The CIA and Pg+CIA groups showed 100% arthritis incidence， with evident joint redness， swelling， and deformity. The number of affected limbs was 27 and 28， and clinical scores were 68 and 70， respectively. No obvious clinical symptoms were observed in the Pg group. Histopathological and imaging analyses showed severe joint lesions in the CIA and Pg+CIA groups， with significantly increased histopathological scores， bone mineral density， bone volume fraction， trabecular thickness， and trabecular number compared to the Control group （

0.01）. No obvious joint pathology was observed in the Pg group. Lung lesions： The Pg+CIA group exhibited marked alveolar inflammation， interstitial inflammatory cell infiltration， and alv

eolar wall thickening， with pronounced blue staining of collagen fibers. Histopathological scores and collagen area ratios were significantly higher than those of the Control， Pg， and CIA groups （

0.05）. Lung protein and mRNA expression levels of

-SMA， ColⅠ， and FN were markedly increased， and mRNA levels of IL-6， TNF-

， and IL-1

were significantly elevated compared to the Control group （

0.05）. Serology： The Pg+CIA group showed significantly higher levels of CCP， Pg， and IgG compared with the Control， Pg， and CIA groups （

0.05）.

Conclusion

DBA/1 mice subjected to Pg infection combined with CIA exhibited pronounced symptoms and pathological features of RA-ILD， along with elevated serum anti-CCP antibody levels. This model represents a novel RA-ILD mouse model， providing a valuable experimental tool for investigating RA-ILD pathogenesis and developing new therapeutics， and serves as a basis for establishing anti-cyclic citrullinated peptide antibody （ACPA）-positive RA-ILD animal models.

关键词

Keywords

references

OLIVEIRA R P ， RIBEIRO R ， MELO L ， et al . Connective tissue disease-associated interstitial lung disease ［J］. Pulmonology ， 2022 ， 28 （ 2 ）： 113 - 118 .

RAIMUNDO K ， SOLOMON J J ， OLSON A L ， et al . Rheumatoid arthritis-interstitial lung disease in the United States：Prevalence，incidence，and healthcare costs and mortality ［J］. J Rheumatol ， 2019 ， 46 （ 4 ）： 360 - 369 .

YAN Q ， HE L ， WANG L ， et al . Experimental animal models for rheumatoid arthritis-associated interstitial lung disease ［J］. Sci Tradit Chin Med ， 2025 ， 3 （ 2 ）： 124 - 136 .

KENNEY H M ， WOOD R W ， RAMIREZ G ， et al . Implementation of automated behavior metrics to evaluate voluntary wheel running effects on inflammatory-erosive arthritis and interstitial lung disease in TNF-Tg mice ［J］. Arthritis Res Ther ， 2023 ， 25 （ 1 ）： 17 .

JENNING M ， MARKLEIN B ， YTTERBERG J ， et al . Bacterial citrullinated epitopes generated by Porphyromonas gingivalis infection-A missing link for ACPA production ［J］. Ann Rheum Dis ， 2020 ， 79 （ 9 ）： 1194 - 1202 .

LUGLI E B ， CORREIA R E S M ， FISCHER R ， et al . Expression of citrulline and homocitrulline residues in the lungs of non-smokers and smokers：Implications for autoimmunity in rheumatoid arthritis ［J］. Arthritis Res Ther ， 2015 ， 17 （ 1 ）： 9 .

NANNINI C ， MEDINA-VELASQUEZ Y F ， ACHENBACH S J ， et al . Incidence and mortality of obstructive lung disease in rheumatoid arthritis：A population-based study ［J］. Arthritis Care Res ， 2013 ， 65 （ 8 ）： 1243 - 1250 .

KIM Y ， YANG H I ， KIM K S . Etiology and pathogenesis of rheumatoid arthritis-interstitial lung disease ［J］. Int J Mol Sci ， 2023 ， 24 （ 19 ）： 14509 .

COURBON G ， RINAUDO-GAUJOUS M ， BLASCO-BAQUE V ， et al . Porphyromonas gingivalis experimentally induces periodontitis and an anti-CCP2-associated arthritis in the rat ［J］. Ann Rheum Dis ， 2019 ， 78 （ 5 ）： 594 - 599 .

JUNG H ， JUNG S M ， RIM Y A ， et al . Arthritic role of Porphyromonas gingivalis in collagen-induced arthritis mice ［J］. PLoS One ， 2017 ， 12 （ 11 ）： e0188698 .

杨超，仲格嘉，刘春芳，等 . 抗瓜氨酸化蛋白抗体高表达类风湿关节炎动物模型的研究进展［J］. 中国组织工程研究， 2022 ， 26 （ 18 ）： 2930 - 2937 .

YANG C ， ZHONG G J ， LIU C F ， et al . Research progress of rheumatoid arthritis animal model with high expression of anti-citrullinated protein antibodies ［J］. Chin J Tissue Eng Res ， 2022 ， 26 （ 18 ）： 2930 - 2937 .

YANG C ， HU Z ， WANG L ， et al . Porphyromonas gingivalis with collagen immunization induces ACPA-positive rheumatoid arthritis in C3H mice ［J］. Clin Immunol ， 2024 ， 258 ： 109859 .

BAI R F ， PEI J Q ， PE S Q ， et al . LPA2 alleviates septic acute lung injury via protective endothelial barrier function through activation of PLC-PKC-FAK ［J］. J Inflamm Res ， 2023 ， 16 ： 5095 - 5109 .

CATRINA A ， KRISHNAMURTHY A ， RETHI B . Current view on the pathogenic role of anti-citrullinated protein antibodies in rheumatoid arthritis ［J］. RMD Open ， 2021 ， 7 （ 1 ）： e001228 .

DIESLER R ， COTTIN V . Pulmonary fibrosis associated with rheumatoid arthritis：From pathophysiology to treatment strategies ［J］. Expert Rev Respir Med ， 2022 ， 16 （ 5 ）： 541 - 553 .

DAI Y ， WANG W ， YU Y ， et al . Rheumatoid arthritis-associated interstitial lung disease：An overview of epidemiology，pathogenesis，and management ［J］. Clin Rheumatol ， 2021 ， 40 （ 4 ）， 1211 - 1220 .

NATALINI J G ， BAKER J F ， SINGH N ， et al . Autoantibody seropositivity and risk for interstitial lung disease in a prospective male-predominant rheumatoid arthritis cohort of U.S.Veterans ［J］. Ann Am Thorac Soc ， 2021 ， 18 （ 4 ）： 598 - 605 .

GROSSE J ， ALLADO E ， ROUX C ， et al . ACPA-positive versus ACPA-negative rheumatoid arthritis：Two distinct erosive disease entities on radiography and ultrasonography ［J］. Rheumatol Int ， 2020 ， 40 （ 4 ）： 615 - 624 .

杨超，胡智星，高双荣，等 . 四种不同抗原诱导的类风湿关节炎小鼠模型的病变特点比较研究［J］. 药学学报， 2023 ， 58 （ 6 ）： 1586 - 1595 .

YANG C ， HU Z ， GAO S ， et al . Comparison of pathological features in mouse models of rheumatoid arthritis induced by four different antigens ［J］. Acta Pharm Sin ， 2023 ， 58 （ 6 ）： 1586 - 1595 .

KRISHNAMURTHY A ， JOSHUA V ， HAJ HENSVOLD A ， et al . Identification of a novel chemokine-dependent molecular mechanism underlying rheumatoid arthritis-associated autoantibody-mediated bone loss ［J］. Ann Rheum Dis ， 2016 ， 75 （ 4 ）： 721 - 729 .

SUN M ， RETHI B ， KRISHNAMURTHY A ， et al . Anticitrullinated protein antibodies facilitate migration of synovial tissue-derived fibroblasts ［J］. Ann Rheum Dis ， 2019 ， 78 （ 12 ）： 1621 - 1631 .

ZHAO F ， ZHANG W D ， JIANG Y L ， et al . Risk factors of patients with interstitial lung disease complicated with rheumatoid arthritis ［J］. J Biol Regul Homeost Agents ， 2020 ， 34 （ 1 ）： 221 - 226 .

AKIYAMA M ， KANEKO Y . Pathogenesis，clinical features，and treatment strategy for rheumatoid arthritis-associated interstitial lung disease ［J］. Autoimmun Rev ， 2022 ， 21 （ 5 ）： 103056 .

Views

150

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Characteristics and Prescription Analysis of Marketed Chinese Patent Medicines for Rheumatoid Arthritis

Animal Models of Functional Constipation： A Review

Danggui Niantongtang in Treatment of Rheumatoid Arthritis： A Review

Guizhi Shaoyao Zhimutang and Active Components of Its Single Herbs in Treatment of Rheumatoid Arthritis： A Review

Clinical and Mechanistic Research Progress on Classical Traditional Chinese Medicine Formulas in Treatment of Chronic Heart Failure

Related Author

LI Siyu

DENG Shuangfei

DING Daiyue

SONG Changyue

SU Xiaohui

KONG Xiangying

HE Youcheng

LIN Shijin

Related Institution

State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs， Institute of Chinese Materia Medica，China Academy of Chinese Medical Sciences

Second People's Hospital Affiliated to Fujian University of Traditional Chinese Medicine（TCM）

Longhua Hospital， Shanghai University of TCM

School of Integrative Medicine， Shanghai University of TCM

West China Hospital of Sichuan University

AI脑图

AI问答

Address：No. 16, Nanxiaojie, Dongzhimennei, Beijing Postal code：100700
Tel：010-84076882 Email：syfjx_2010@188.com
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备11006657号-10 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰