南方医科大学学报

南方医科大学学报 ›› 2022, Vol. 42 ›› Issue (6): 860-867.doi: 10.12122/j.issn.1673-4254.2022.06.09

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血管紧张素转化酶2参与臭氧暴露导致的小鼠肺部炎症及气道重塑

王月霞，张 钰，张 亮，李梦圆，朱培育，纪望全，梁若楠，秦露伟，吴卫东，冯斐斐，晋乐飞   

  1. 郑州大学公共卫生学院流行病学教研室，毒理学教研室，河南 郑州 450001；河南省疾病预防控制中心慢性非传染性疾病防治研究所，河南 郑州 450001；新乡医学院公共卫生学院劳动卫生与环境卫生学教研室，河南 新乡 453000
  • 出版日期:2022-06-20 发布日期:2022-06-27

Angiotensin-converting enzyme 2 particapates in ozone-induced lung inflammation and airway remodeling in mice

WANG Yuexia, ZHANG Yu, ZHANG Liang, LI Mengyuan, ZHU Peiyu, JI Wangquan, LIANG Ruonan, QIN Luwei, WU Weidong, FENG Feifei, JIN Yuefei   

  1. Department of epidemiology, Department of Toxicology, School of Public Health, Zhengzhou University, Zhengzhou 450001, China; Institute of Chronic and Non-communicable Disease Prevention and Control, Henan Provincial Center for Disease Control and Prevention, Zhengzhou 450001, China; Department of Occupational and Environmental Health, School of Public Health, Xinxiang Medical University, Xinxiang 453000, China
  • Online:2022-06-20 Published:2022-06-27

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摘要： 目的 研究血管紧张素转换酶2（ACE2）在臭氧暴露致小鼠肺部炎症及气道重塑中的作用。方法 16只野生型（WT）C57BL/6J小鼠和16只ACE2 敲除（KO）小鼠分别暴露于空气和臭氧（0.8 ppm）中，每天暴露3 h，连续暴露5 d。分别设置AirWT组，Air KO组，Ozone WT组和Ozone KO组。Masson染色观察小鼠肺组织胶原沉积情况，HE染色观察小鼠肺部病理学改变，并进行评分；收集小鼠支气管肺泡灌洗液（BALF），并进行细胞计数；分别采用BCA法和酶联免疫吸附法（ELISA）测定BALF总蛋白和细胞因子浓度；荧光定量PCR检测肺组织中气道重塑相关指标的mRNA表达；小鼠的气道阻力采用小动物呼吸机测量（乙酰甲胆碱激发）。结果 臭氧暴露ACE2 KO小鼠肺组织炎症病理学变化总分显著高于臭氧暴露WT小鼠（P<0.05）。Masson染色结果显示，臭氧暴露小鼠肺组织支气管和肺泡周围有大量的胶原沉积，臭氧暴露ACE2 KO小鼠肺组织的支气管和肺泡的胶原沉积阳性区域面积为（19.62±3.16）%、（21.63±3.78）%，高于臭氧暴露 WT 小鼠肺组织的（6.49±1.34）%、（4.44± 0.99）%，两者差异具有统计学意义（P<0.05）。臭氧暴露ACE2 KO小鼠BALF中总细胞数与总蛋白含量均高于臭氧暴露WT小 鼠，差异无统计学意义（P>0.05）。臭氧暴露ACE2 KO小鼠BALF中IL-6、IL-1β、TNF-α、趋化因子（C-X-C基序）配体1（CXCL1/KC）和单核细胞趋化蛋白（MCP-1）水平均高于臭氧暴露WT小鼠，其中IL-1β水平的变化差异具有统计学意义（P<0.05）。臭氧暴露 ACE2 KO 小鼠的肺组织中的基质金属蛋白酶 9（MMP-9）、MMP-13、金属蛋白酶组织抑制剂（TIMP4）、Ⅰ型胶原α1 （COL1A1）、TGF-β的mRNA水平均明显高于WT组小鼠，两组的MMP-9、TIMP4、COL1A1、TGF-β的mRNA水平差异具有统计学意义（P<0.01）。两组小鼠在0、10、25、100 mg/mL的乙酰甲胆碱浓度下的气道阻力的差异无统计学意义（P>0.05）。结论 ACE2参与臭氧暴露导致的小鼠肺部炎症及气道重塑。

关键词: 血管紧张素转换酶2；臭氧；炎症；气道重塑

Abstract: Objective To investigate the roles of angiotensin-converting enzyme 2 (ACE2) in ozone-induced pulmonary inflammation and airway remodeling in mice. Methods Sixteen wild-type (WT) C57BL/6J mice and 16 ACE2 knock-out (KO) mice were exposed to either filtered air or ozone (0.8 ppm) for 3 h per day for 5 consecutive days. Masson's staining and HE staining were used to observe lung pathologies. Bronchoalveolar lavage fluid (BALF) was collected and the total cell count was determined. The total proteins and cytokines in BALF were determined by BCA and ELISA method. The transcription levels of airway remodeling-related indicators in the lung tissues were detected using real-time quantitative PCR. The airway resistance of the mice was measured using a small animal ventilator with methacholine stimulation. Results Following ozoneexposure ACE2 KO mice had significantly higher lung pathological scores than WT mice (P<0.05). Masson staining results showed that compared with ozone-exposed WT mice, ozone-exposed ACE2 KO mice presented with significantly larger area of collagen deposition in the bronchi [(19.62±3.16)% vs (6.49±1.34)%, P<0.05] and alveoli [(21.63±3.78)% vs (4.44±0.99)%, P<0.05]. The total cell count and total protein contents in the BALF were both higher in ozone-exposed ACE2 KO mice than in WT mice, but these differences were not statistically significant (P>0.05). The concentrations of IL-6, IL-1β, TNF-α, CXCL1/KC and MCP-1 in the BALF were all higher in ozone-exposed ACE2 KO mice than in ozone-exposed WT mice, but only the difference in IL-1β was statistically significant (P<0.05). The transcription levels of MMP-9, MMP-13, TIMP 4, COL1A1, and TGF-β in the lung tissues were all significantly higher in ozone-exposed ACE2 KO mice (P<0.01). No significant difference was found in airway resistance between ozone-exposed ACE KO mice and WT mice after challenge with 0, 10, 25, or 100 mg/mL of methacholine. Conclusion ACE2 participates in ozone-induced lung inflammation and airway remodeling in mice.

Key words: angiotensin-converting enzyme 2; ozone; lung inflammation; airway remodeling