新生期小鼠肺炎链球菌肺炎对气道上皮损伤的影响  

Effects of neonatal Streptococcus pneumoniae pneumonia on airway epithelial injury of mice

作　　者：吴毅 孔潇 李沁原 陈诗懿 李媛媛[2] 张光莉[2] 田小银 罗征秀[2] WU Yi;KONG Xiao;LI Qin-yuan;CHEN Shi-yi;LI Yuan-yuan;ZHANG Guang-li;TIAN Xiao-yin;LUO Zhengxiu(Institute of Pediatrics,Children's Hospital Affiliated to Chongqing Medical University,Key Laboratory of Child Development Disease Research,Ministry of Education,National Center for Clinical Medical Research on Child Health and Diseases,National International Scientific and Technological Cooperation Base for Major Diseases of Child Development,Chongqing Key Laboratory of Pediatrics,Chongqing 400014,China;Department of Respiratory Medicine,Children’s Hospital Affiliated to Chongqing Medical University,Chongqing 400014,China)

机构地区：[1]重庆医科大学附属儿童医院儿科研究所/儿童发育疾病研究教育部重点实验室/国家儿童健康与疾病临床医学研究中心/儿童发育重大疾病国家国际科技合作基地/儿科学重庆市重点实验室,重庆400014 [2]重庆医科大学附属儿童医院呼吸科,重庆400014

出　　处：《解放军医学杂志》2019年第11期919-924,共6页Medical Journal of Chinese People's Liberation Army

基　　金：重庆市科委重点项目(cstc2017jcyjBX0049);重庆市渝中区科委项目(20170120)~~

摘　　要：目的探讨新生期肺炎链球菌肺炎(S.pp)对气道上皮损伤的影响。方法生后7 d的C57BL/6小鼠鼻腔内滴注2×10^5 cfu肺炎链球菌(5μl)建立S.pp模型,对照组滴注等量磷酸盐缓冲液。感染5周后检测两组小鼠的气道高反应性,收集小鼠肺组织,利用实时荧光定量PCR检测CDH1、TJP1 mRNA水平;免疫组织化学、Western blotting检测气道上皮E钙黏蛋白(E-cadherin)、紧密连接蛋白(ZO-1)水平。Masson染色检测小鼠气道上皮下胶原蛋白沉积水平,ABPAS染色检测小鼠气道杯状细胞数量,HE染色检测小鼠肺组织的炎症改变情况。结果小鼠雾化吸入乙酰甲胆碱浓度为6.25~50 mg/ml时,S.pp组小鼠的气道高反应性明显高于对照组(P<0.001)。S.pp组小鼠肺组织CDH1、TJP1 mRNA水平明显低于对照组(分别为0.85±0.29 vs.1.42±0.40,P=0.033;0.43±0.16 vs.0.83±0.26,P=0.010),气道上皮细胞E-cadherin、ZO-1的表达水平明显低于对照组(分别为9.66±4.89 vs.24.52±7.58,P=0.001;13.54±3.79 vs.25.53±5.99,P=0.005),肺组织E-cadherin、ZO-1蛋白水平明显低于对照组(分别为0.23±0.06 vs.0.38±0.06,P=0.019;0.68±0.12 vs.0.96±0.16,P=0.032),上皮下胶原蛋白沉积与对照组比较明显增加(45.54±5.79 vs.26.3±5.53,P=0.001)。两组气道杯状细胞数量及肺组织炎症评分差异无统计学意义。结论S.pp可诱导小鼠气道上皮损伤,参与形成气道高反应性。Objective To investigate the effects of neonatal Streptococcus pneumoniae pneumonia(S.pp)on airway epithelial injury of mice.Methods Neonatal C57 BL/6(1-week-old)mice were infected intranasally with 2×105 cfu of Streptococcus pneumoniae(S.p)in a volume of 5μl sterile phosphate buffered saline(PBS)(S.pp group),while the mock-infected controls received the same volume of sterile PBS(control group).Five weeks after the infection,the airway hyperresponsiveness(AHR)was evaluated by invasive body plethysmography system.The lung tissue was harvested,CDH1 and TJP1 mRNA levels were analyzed by RT-PCR.Immunohistochemical method and Western blotting were used to evaluate the expressions of E-cadherin and tight junction protein(ZO-1).Airway inflammation was detected by HE staining,the subcutaneous collagen deposition beneath the airway epithelium was evaluated by Masson staining,and the number of goblet cells was detected by Alcian Blue Periodic Acid Schiff(AB-PAS)staining.Results When the concentration of aerosolized methacholine inhaled by mice ranged from 6.25 to 50 mg/ml,the AHR was obviously higher in S.pp group than that in control group(P<0.001);the levels of CDH1 and TJP1 mRNA in the lung tissue were markedly lower in S.pp group(0.85±0.29 and 0.43±0.16)than those in control group(0.85±0.29 vs.1.42±0.40,P=0.033;0.43±0.16 vs.0.83±0.26,P=0.010,respectively);the expression levels of E-cadherin and ZO-1 in airway epithelium were lower in S.pp group than those in control group(9.66±4.89 vs.24.52±7.58,P=0.001;13.54±3.79 vs.25.53±5.99,P=0.005,respectively);the protein levels of E-cadherin and ZO-1 in lung tissue were remarkably lower in S.pp group than those in control group(0.23±0.06 vs.0.38±0.06,P=0.019;0.68±0.12 vs.0.96±0.16,P=0.032,respectively);the subcutaneous collagen deposition beneath the airway epithelium increased significantly in S.pp group than that in control group(45.54±5.79 vs.26.3±5.53,P=0.001).The number of goblet cells and inflammatory cell infiltration showed no statistical significance

关 键 词：肺炎链球菌肺炎 新生期 气道上皮 支气管高反应性 

分 类 号：R563.1[医药卫生—呼吸系统]