摘要
目的:构建一种模拟人类类风湿关节炎(rheumatoid arthritis,RA)并间质性肺疾病(RA-associated interstitial lung disease,RA-ILD)病理特征的RA-ILD大鼠模型。方法:将24只SD大鼠随机分为3组,分别为对照组、RA组和RA-ILD组,每组各8只。行胶原诱导性关节炎(collagen-induced arthritis,CIA)造模,RA-ILD组同时采用气管内滴入博来霉素(bleomycin,BLM)行肺纤维化造模。观测大鼠体质量并进行关节炎评分;ELISA检测血清类风湿因子(rheumatoid factor,RF)、抗环瓜氨酸肽抗体(anti-cyclic citrullinate peptide antibody,Anti-CCP-Ab)、C反应蛋白(C-reactive protein,CRP)、基质金属蛋白酶7(matrix metalloproteinase-7,MMP-7)、转化生长因子β1(transforming growth factor-β1,TGF-β1)和肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)含量;HE及Masson染色观察滑膜和肺组织病理学形态改变,依据染色结果评判大鼠RA-ILD发病率;免疫组化和Western blot方法检测肺组织MMP-7和白细胞介素22(interleukin-22,IL-22)表达水平。结果:(1)与对照组比,造模第0、7、14、21和28天,RA组和RA-ILD组大鼠体质量无显著差异,第35天时RA组和RA-ILD组大鼠体质量显著降低(P<0.05);(2)与对照组相比,第7、14、21、28和35天时RA组和RA-ILD组关节炎评分均显著升高(P<0.05)。(3)ELISA结果显示,构建RAILD4周(37 d)后,RA组和RA-ILD组血清RF、Anti-CCP-Ab、TNF-α、MMP-7、TGF-β1及CRP含量均较对照组显著升高(P<0.05);RA-ILD组血清Anti-CCP-Ab和TNF-α含量较RA组显著升高(P<0.05)。(4)构建RA-ILD模型4周后,HE染色发现RA组和RA-ILD组滑膜均出现明显的增生及炎症浸润现象;肺部皆可见明显肺泡结构紊乱,肺泡间隔增宽,肺泡壁增厚,大量炎性细胞渗出。构建RA-ILD模型4周后,Masson染色观察到明显的蓝色胶原纤维沉积及纤维灶形。(5)构建RA-ILD模型4周后,与对照组相比,RA组和RA-ILD组大鼠RA-ILD发病率显著升高(P<0.05);与RA组相比,RA-ILD组RA-ILD发病率显著升高(P<0.05)。(6)免疫组化和Western blot结果显示,构建RA-ILD模型4周后,与对照组相比,RA组和RA-ILD组肺组织MMP-7和IL-22表达显著上调。结论:SD大鼠经CIA联合BLM诱导建立的RA-ILD模型可作为研究RA-ILD发病机制的一种实验动物模型。
AIM:To establish a rat model of rheumatoid arthritis(RA)-associated interstitial lung disease(RA-ILD)that mimics the pathological features of human RA with interstitial pneumonia.METHODS:Twenty-four SD rats were randomly divided into 3 groups:control group,RA group and RA-ILD group.The collagen-induced arthritis(CIA)model was established in RA group and RA-ILD group.The mice in RA-ILD group were treated with bleomycin(BLM)intratracheal instillation for modeling of pulmonary fibrosis.The body weight and arthritis scores of the rats were observed.Serum levels of rheumatoid factor(RF),anti-cyclic citrullinate peptide antibody(Anti-CCP-Ab),C-reactive protein(CRP),matrix metalloproteinase-7(MMP-7),transforming growth factor-β1(TGF-β1)and tumor necrosis factor-α(TNF-α)were measured by ELISA.Histopathological changes of the synovium and lung were observed by HE and Mas‐son staining.The incidence of RA-ILD in the rats was evaluated according to the results of HE and Masson staining.The protein expression levels of MMP-7 and interleukin-22(IL-22)in lung tissues were measured by immunohistochemistry and Western blot.RESULTS:(1)Compared with control group,there was no significant difference in body weight be‐tween RA group and RA-ILD group on day 0,day 7,day 14,day 21 and day 28,while the body weight of the rats in RA group and RA-ILD group on day 35 was lower than that in control group(P<0.05).(2)Compared with control group,the arthritis scores of the rats in RA group and RA-ILD group on day 7,day 14,day 21,day 28 and day 35 were significantly increased(P<0.05).(3)ELISA results showed that the serum levels of RF,Anti-CCP-Ab,TNF-α,MMP-7,TGF-β1 and CRP in RA and RA-ILD groups were significantly increased compared with control group(P<0.05),and the serum levels of Anti-CCP-Ab and TNF-αin RA-ILD group were significantly higher than those in RA group on day 37 after the construction of RA-ILD for 4 weeks(P<0.05).(4)HE staining showed that the synovium of the rats in RA group and RA-ILD group showed obvious synovial hyperplasia and inflammatory infiltration on day 37.The rat lungs in RA group and RA-ILD group showed obvious alveolar structural disorder,widened alveolar septum,thickened alveolar wall,and mas‐sive inflammatory cell exudation.On day 37,Masson staining showed obvious blue collagen fiber deposition and fiber foci in lungs of RA group and RA-ILD group.(5)On day 37,the incidence of RA-ILD in RA and RA-ILD groups was higher than that in control group(P<0.05),and that in RA-ILD group was higher than that in RA group(P<0.05).(6)Immu‐nohistochemistry and Western blot results showed that the expression levels of MMP-7 and IL-22 in lung tissues of the rats in RA group and RA-ILD group on day 37 were significantly up-regulated compared with control group.CONCLUSION:The RA-ILD model of SD rats induced by CIA combined with BLM can be used as an experimental animal model to study the pathogenesis of RA-ILD.
作者
欧慧萍
吴趋荟
袁多
柳玉佳
林辉
李鑫
范伏元
OU Huiping;WU Quhui;Yuan Duo;LIU Yujia;LIN Hui;LI Xin;FAN Fuyuan(Hunan University of Chinese Medicine,Changsha 410208,China;Hunan Traditional Chinese Medicine College,Zhu-zhou 412012,China;The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410007,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2023年第6期1145-1152,共8页
Chinese Journal of Pathophysiology
基金
国家自然科学基金项目(No.81803993)
湖南省自然科学基金项目(No.2020JJ5434)
湖南省教育厅科学研究项目(No.19C1446)
湖南省中医药科研计划项目(No.2021038)
湖南中医药高等专科学校校级科研项目(No.XY2022.006)。
