摘要
研究博来霉素(BLM)致小鼠肺纤维化模型中MMP-7、KL-6、SP-A、SP-D的表达和临床意义,探讨靶向性近红外荧光MMP-7探针及4种生物标志物联合检测在肺纤维化模型中的应用,旨在为肺纤维化的诊断、预后、动态监测及治疗提供参考。40只雄性C57BL/6小鼠随机分为BLM模型组和对照组,模型组气管滴注100μL低(2.0 mg·kg^(-1))、中(3.5 mg·kg^(-1))、高(5.0 mg·kg^(-1))剂量的盐酸博来霉素注射液建立肺纤维化模型,依次记为L组、M组和H组,对照组给予等量生理盐水。于建模后28 d,进行肺组织肺泡炎和肺纤维化评分,使用ELISA及RT-PCR技术检测MMP-7、KL-6、SP-A、SP-D的表达量,免疫组化法和Western blot检测MMP-7和KL-6蛋白在肺组织中的表达;腹腔注射近红外荧光MMP-7探针,于1.0、1.5、2.0、12.0、24.0、48.0 h进行荧光活体成像,观察MMP-7探针在体内的动态分布。结果表明,BLM摄入剂量与肺泡炎(P=0.008)及肺纤维化程度(P=0.03)显著相关(P<0.05), 2.0、3.5、5.0 mg·kg^(-1)剂量的BLM在一定程度上均能引起MMP-7、KL-6、SP-A、SP-D表达异常(P<0.05);近红外荧光MMP-7探针能特异性结合MMP-7靶点,探针注射12 h后靶向分布于肺部,信号稳定。4种生物标志物在肺纤维化模型小鼠肺组织中的表达随肺泡炎或纤维化的严重程度而增高或降低,提示它们是肺纤维化的生物标志物,联合检测可以提高方法的准确率和敏感性,非侵入性的MMP-7探针可用于小鼠肺纤维化的动态监测。
Pulmonary fibrosis(PF)is a disease that with high mortality,poor prognosis and unclear pathogenesis.To investigate the expression of MMP-7,KL-6,SP-A,SP-D in bleomycin-induced PF mice,to discuss the application of target-specific near-infrared fluorescent MMP-7 probe and combined detection of 4 markers in bleomycin-induced PF mice to provide the foundation of diagnosis,progress and prognosis of the disease.Forty C57BL/6 male mice were randomly divided into bleomycin group(n=30)and control group(n=10),the different concentrations(2.0,3.5,5.0 mg·kg-1)of bleomycin were given via trachea to induce the pulmonary fibrosis model in bleomycin group,the control group were given the same dose of saline.At day 28 after administration,histopathological score was use to evaluated the severity of pulmonary fibrosis and alveolitis.The expressions of MMP-7,KL-6,SP-A,SP-D were detected by ELISA,RT-PCR,immunohistochemistry and Western blot.The mice were subjected to NIR imaging at different time after injection of MMP-7 probe to observed the distribution and targeting effect of the probes in vivo.The concentration of BLM was correlated with alveolitis pathological score(P=0.008)and fibrosis pathological score(P=0.03).The expressions of MMP-7,KL-6,SP-A,SP-D were significantly different between bleomycin group and control group(P<0.05).Near-infrared fluorescence probe can specifically target on MMP-7 with stable signal.The abnormal expressions of MMP-7,KL-6,SP-A,SP-D were correlated with PF,they may be a biomarkers of bleomycin-induced pulmonary fibrosis.The combined detection of these four markers can improve the diagnosis ability in PF and provide a strong reliability in diagnosis in clinical use.Noninvasive MMP-7 probe can be used on the PF disease of dynamic monitoring.
作者
王国辽
张洁
饶家榕
莫睿文
远立国
WANG Guoliao;ZHANG Jie;RAO Jiarong;MO Ruiwen;YUAN Liguo(College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China;Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases,Guangzhou 510642,China)
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2021年第4期1134-1140,共7页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
广东省自然科学基金(2017A030313182)。
作者简介
同等贡献作者:王国辽(1994-),男,黎族,海南白沙人,硕士生,主要从事兽医外科研究,E-mail:wangguoliao@stu.scau.edu.cn;同等贡献作者:张洁(1991-),女,新疆哈密人,硕士生,主要从事兽医外科研究,E-mail:1390256148@qq.com;通信作者:远立国,主要从事兽医外科研究,E-mail:yuanliguo@scau.edu.cn。
