氧化铍致大鼠肺纤维化的实验研究  

Experimental Study on Pulmonary Fibrosis Induced by Beryllium Oxide in Rats

作　　者：王凯 李思睿 王宇欣 蔺文轩 刘志宏 WANG Kai;LI Sirui;WANG Yuxin;LIN Wenxuan;LIU Zhihong(School of Public Health and Management,Ningxia Medical University,Yinchuan750004,China)

机构地区：[1]宁夏医科大学公共卫生与管理学院

出　　处：《宁夏医科大学学报》2019年第11期1091-1095,共5页Journal of Ningxia Medical University

基　　金：国家自然科学基金(81360415);宁夏自然科学基金(2019AAC03085)

摘　　要：目的探讨氧化铍(BeO)致SD大鼠肺纤维化过程中肺组织的病理和超微结构的变化以及I型胶原(COL-I)、III型胶原(COL-Ⅲ)、α-平滑肌肌动蛋白(α-SMA)的表达。方法64只SPF级健康雄性SD大鼠,随机分为生理盐水组(32只)和BeO染毒组(32只)。BeO染毒组气管灌注BeO混悬液0.5 mL(浓度为10mg·mL^-1),生理盐水组注入等量的0.9%无菌生理盐水。两组大鼠分别于染毒后的第20、40、60、80天取样,每组各时间点分别处死8只大鼠。取肺组织制备光学显微镜和电子显微镜病理切片观察肺组织病理变化,并采用实时定量PCR(qRT-PCR)法和免疫印迹法(Western blot)分别检测COL-I、COL-III和α-SMA基因和蛋白水平。结果Masson染色可观察到染毒后支气管及肺泡间隔有蓝色胶原沉积,随着时间的延长蓝色胶原增加。电子显微镜可观察到染毒后第20、40天后肺泡Ⅱ型上皮细胞胞浆嗜锇性板层小体排空;第60、80天后肺泡Ⅱ型上皮细胞胞浆嗜锇性板层小体致密,肺间质可见纤维束存在。BeO染毒后COL-I mRNA和蛋白表达升高且随时间延长而增加。BeO染毒第60、80天COL-III mRNA和蛋白表达水平升高,且随着时间延长而增加。BeO染毒后α-SMA mRNA和蛋白表达升高;α-SMA mRNA随着时间延长而增加。结论BeO可诱导肺组织发生纤维化,细胞外基质成分COL-I、COL-III及肺成纤维细胞转分化标记物α-SMA在肺组织中表达升高。Objective To investigate the pathological changes and ultrastructural changes of lung tissue and expression of COL-I、CLOL-III and a-SMA during pulmonary fibrosis induced by beryllium oxide(BeO)in SD rats.Methods Sixty-four SPF healthy male SD rats were randomly divided into normal saline group and BeO-treated group.The BeO-treated group was instilled with 0.5 mL of BeO suspension(concentration:10 mg·mL^-1)and the normal saline group was injected with an equal amount of 0.9%sterile physiological saline.The two groups of rats were respectively neutralized at 20,40,60,and 80 d after exposure,eight rats in each group were killed at each time point.The pathological changes of lung tissue were observed by light microscopy and electron microscopy.The gene and protein levels of COL-I,CLOL-III andα-SMA were detected by Real-time quantitative PCR(qRT-PCR)and Western blot.Results Masson staining showed that blue collagen deposition was observed in the bronchi and alveolar spaces after exposure.The cytoplasmic eosinophilic lamellar corpuscles of alveolar type II epithelial cells were observed by electron microscopy after exposure for 20 and 40 days;the cytoplasmic eosinophilic lamellar bodies of lung alveolar type II epithelial cells were dense after exposure for 60 and 80 days and fibrous bundle was present in the pulmonary interstitial.In the BeO-treated group,COL-I mRNA and protein expression increased and increased with time.The expression of COL-III mRNA and protein was increased at 60 and 80 d after BeO exposure and increased with time.The expression ofα-SMA mRNA and protein increased after BeO exposure;α-SMA mRNA increased with time.Conclusion BeO can induce fibrosis in lung tissue,and the expression of extracellular matrix components COL-I and COL-III is increased in lung tissue and the expression ofα-SMA is increased.

关 键 词：氧化铍 肺纤维化 I型胶原 III型胶原 Α-平滑肌肌动蛋白 

分 类 号：R563[医药卫生—呼吸系统]