大鼠口腔溃疡模型构建与伦理优化研究  

Establishement and ethical optimization of rat oral mucosa ulcer model

作　　者：李想 王凯燕 于玮玮 郝心仪 李玲 段建红 冯斌 刘青 夏凌云[1] 牛丽娜 LI Xiang;WANG Kaiyan;YU Weiwei;HAO Xinyi;LI Ling;DUAN Jianhong;FENG Bin;LIU Qing;XIA Lingyun;NIU Lina(Stomatology Center,Taihe Hospital,Hubei University of Medicine,China,442000 Shiyan;State Key Laboratory of Oral&Maxillofacial Reconstruction and Regeneration,National Clinical Research Center for Oral Diseases,Shaanxi Key Laboratory of Stomatology,The Third Affiliated Hospital of Air Force Medical University,Xi'an)

机构地区：[1]湖北医药学院附属太和医院口腔医学中心,十堰442000 [2]口颌系统重建与再生全国重点实验室,国家口腔疾病临床医学研究中心,陕西省口腔医学重点实验室,空军军医大学第三附属医院

出　　处：《实用口腔医学杂志》2025年第1期26-33,共8页Journal of Practical Stomatology

基　　金：国家杰出青年科学基金(编号:82325012)。

摘　　要：目的:通过不同的实验方法分别构建大鼠口腔黏膜溃疡模型,以满足研究需求并体现动物伦理关怀。方法:将96只SD大鼠随机分组,对照组(n=8)大鼠常规饲养无其他处理,冰醋酸烧灼组分别采用20%、40%、60%浓度的冰醋酸处理颊黏膜(n=8);摩擦损伤组用高速车针(30000 r/min)摩擦颊黏膜,根据损伤面积分为10、20、30 mm^(2)3组(n=8);电离辐照组分别采用10、12、15、20、30 Gy的辐照剂量照射颊黏膜(n=8)。分别处理后采用HE、Masson以及免疫荧光染色技术检测黏膜组织病变情况,qPCR和ELISA检测局部及血清关键炎症因子的变化,观察记录大鼠饮食、体重等机体状况,用Morton&Griffiths五项评估法分析动物疼痛、痛苦与不适。结果:40%和60%的冰醋酸、20 mm^(2)和30 mm^(2)的摩擦损伤、12 Gy以上剂量的电离辐照均可造成87.5%以上受试大鼠口腔黏膜溃疡,溃疡病程时间可持续6~7 d,溃疡局部组织TNF-α和IL-1βmRNA表达升高。血清TNF-α和IL-1β蛋白表达升高。5项评估法显示40%冰醋酸、20 mm^(2)摩擦损伤、12 Gy辐照刺激下大鼠疼痛、痛苦与不适计分最低。结论:40%浓度的冰醋酸、20 mm^(2)的摩擦损伤Gy的电离辐照均可以构建稳定的溃疡模型,且更符合动物实验中的3R原则。Objective:To develop rat models of oral mucosa ulcer using distinct experimental methodologies,fulfilling research requirements and adhering to the ethical standards for animal care.Methods:96 SD rats were randomly allocated into groups.The rats in control group(n=8)were regularly fed without other treatment.Those in chemical cauterization groups were treated by 20%,40%,60%of glacial acetic acid(GAA)on oral mucosa(n=8);in mechanical damage groups by 30000 r/min high speed drill induced trauma of 10,20 and 30 mm^(2) respectively(n=8);in ionizing radiation groups were treated with 10,12,15,20 and 30 Gy on the mucosa respectively(n=8).After the ulcer was appeared,the morphology of the mucosa were observed,the mucosal tissue lesions were examined by HE,Masson and immunofluorescence staining,the expression of TNF-αand IL-1βwere detected by qPCR and ELISA respectively,and the body conditions such as diet and body weight of the rats were observed,the pain,distress and discomfor of the rats were evaluated by MORTON&Griffits Guidelines.Results:40%and 60%GAA,20 mm^(2) and 30 mm^(2) friction damage and ionizing radiation of 12 Gy or greater may induce oral mucosa ulcer with a diseas coruse of 6-7 d in SD rats.TNF-αand IL-1βmRNA expression in the damaged tissue,the related protein expression in blood serum of the rats were increased.MORTON&Griffits Guidelines analysis showed 40%GAA,20 mm^(2) friction damage and 12 Gy ionizing radiation induced the lowest scores of pain,distress and discomfort of the rats with compatible oral mocosa ulcere induced by the relevat treatment.Conclusion:40%GAA,20 mm^(2) of friction damage and 12 Gy of ionizing radiation can reliably establish oral mucosa ulcer models and minimize adverse effects on SD rats,and accord with ethical standards of 3R for laboratory animal.

关 键 词：口腔溃疡 动物模型 伦理关怀 

分 类 号：R781.55[医药卫生—口腔医学]