摘要
目的 探究链脲佐菌素(streptozotocin, STZ)诱导糖尿病肺纤维化(diabetic pulmonary fibrosis, DPF)小鼠模型的建立方法,为临床研究提供稳定的DPF动物模型。方法 将60只雄性C57BL/6小鼠随机分为3组:正常对照组(NG,n=20)、糖尿病肺纤维化1组(DPF1,n=20)、糖尿病肺纤维化2组(DPF2,n=20)。隔夜禁食不禁水后测定小鼠空腹血糖和体重,随后DPF1组一次性腹腔注射大剂量STZ(150 mg/kg),DPF2组连续5 d每天腹腔注射小剂量STZ(50 mg/kg),NG组腹腔注射等量无菌柠檬酸盐缓冲液。注射完毕后,每天观察小鼠的一般情况,每周测定小鼠体重和随机血糖(random blood glucose, RBG),正常喂养16周,每4周每组随机选取5只小鼠处死取材,行病理和分子生物学检测,评估小鼠肺纤维化的程度。结果 STZ诱导后,DPF1组和DPF2组均出现“多饮、多尿、多食、体重减轻(三多一少)”的典型糖尿病症状。DPF1组和DPF2组诱导后体重增加均显著低于NG组,并于第8周后缓慢减轻(P<0.05);DPF1组和DPF2组随机血糖分别于第1周和第2周后高于16.7 mmol/L,并于第9周后趋于稳定(P<0.05)。病理染色结果显示,与NG组相比,DPF1组和DPF2组4周时均未见到明显纤维化病变;8周时可于血管周围见少量纤维化病变;12周时可见明显纤维化病变,且主要累及血管周围;16周时可见更明显的纤维化病变,且已累及周围肺组织。分子生物学结果与其相似,与NG组相比,DPF1组和DPF2组4周和8周时小鼠纤维化标志物胶原蛋白3的表达水平未见明显变化;12周时胶原蛋白3的表达明显增加(P<0.05);16周时胶原蛋白3的表达较12周时增加更明显(P<0.05)。结论 单次大剂量(150 mg/kg)腹腔注射STZ和连续5次小剂量(50 mg/kg)腹腔注射STZ经16周饲养后均可诱导DPF模型,均是DPF的理想动物模型。
Objective To explore methods of streptozotocin⁃induced diabetic pulmonary fibrosis mouse model establishment and provide a stable animal model of diabetic pulmonary fibrosis for clinical research.Methods Sixty male C57BL/6 mice were randomly divided into three groups:normal control(NG,n=20),diabetic pulmonary fibrosis 1(DPF1,n=20),and diabetic pulmonary fibrosis 2(DPF2,n=20)groups.The fasting blood glucose levels and body weight of the mice were measured after overnight fasting;then the DPF1 group was intraperitoneally injected with a large dose of streptozotocin(150 mg/kg),and the DPF2 group was intraperitoneally injected with a small dose of streptozotocin(50 mg/kg)every day for five consecutive days.The NG group was intraperitoneally injected with the same amount of sterile citrate buffer.After injection,the general condition of the mice was observed every day,and the body weight and random blood glucose of the mice were measured every week.The mice were fed normally for 16 weeks,and five mice were randomly selected from each group every 4 weeks to be sacrificed for pathology and molecular biology assays to assess the degree of pulmonary fibrosis.Results After streptozotocin induction,both the DPF1 and DPF2 groups had typical symptoms of diabetes including polydipsia,polyuria,polyphagia and weight loss.The body weight of the DPF1 and DPF2 groups increased slower than that of the NG group,and gradually decreased from the 8th week(P<0.05).The random blood glucose of the DPF1 and DPF2 groups was greater than 16.7 mmol/L after the 1st week and the 2nd week,respectively,and tended to be stable after the 9th week(P<0.05).The pathological results showed that,compared with the NG group,the DPF1 and DPF2 groups had no obvious fibrotic lesions at 4 weeks.However,a small number of fibrotic lesions were observed around the vessels at 8 weeks.Obvious fibrotic lesions were observed at 12 weeks,mainly accumulated around the blood vessels;and more obvious fibrotic lesions were observed at 16 weeks that involved the surrounding lung tissue.The molecular biological results were similar to the pathological results.Compared with the NG group,the DPF1 and DPF2 groups’expression levels of mouse fibrosis marker collagen 3 did not change significantly at 4 and 8 weeks,but increased at 12 weeks(P<0.05)and significantly increased at 16 weeks(P<0.05).Conclusions A single high⁃dose intraperitoneal injection of streptozotocin(150 mg/kg)and five consecutive low⁃dose intraperitoneal injections of streptozotocin(50 mg/kg)induced diabetic pulmonary fibrosis mouse models after 16 weeks of feeding,and both produced ideal animal models for diabetic pulmonary fibrosis.
作者
付庭吕
李宁
刘博昊
熊锐
何如愿
卢子龙
李国瑞
耿庆
FU Tinglyu;LI Ning;LIU Bohao;XIONG Rui;HE Ruyuan;LU Zilong;LI Guorui;GENG Qing(Renmin Hospital of Wuhan University,Wuhan 430060,China)
出处
《中国实验动物学报》
CAS
CSCD
北大核心
2023年第2期194-200,共7页
Acta Laboratorium Animalis Scientia Sinica
基金
国家自然科学基金(8210082163)
中央高校基本科研专项基金(2042021kf0081)。
