小鼠气道Club细胞发育特征分析  

Study on the development characteristics of airway Club cells in mice

作　　者：简鼎 王艺颖 朱琳[1] 李媛媛[1,2] 张光莉[2] 陈诗懿 罗征秀[2] Jian Ding;Wang Yi-Ying;Zhu Lin;Li Yuan-Yuan;Zhang Guang-Li;Chen Shi-Yi;Luo Zheng-Xiu(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》2022年第4期340-345,共6页Medical Journal of Chinese People's Liberation Army

基　　金：重庆市自然科学基金面上项目(cstc2020jcyj-msxmX0677);重庆市渝中区科委项目(20200156);儿童发育疾病研究教育部重点实验室基础研究一般项目(GBRP-202112)。

摘　　要：目的探讨小鼠气道Club细胞的发育特征。方法收集新生期(1周龄)、幼年期(3周龄)、成年期(6周龄)BALB/c小鼠的肺组织及血清,采用免疫荧光检测主支气管和细支气管Club细胞的分布及表达丰度;流式细胞术检测气道上皮Club细胞百分比及增殖能力;ELISA法检测肺匀浆、血清中Club细胞分泌蛋白(CC16)水平。结果各鼠龄期主支气管Club细胞表达位置参差不齐,细支气管Club细胞分布形态平坦且均匀。随着鼠龄的增长,主支气管Club细胞平均荧光强度(MFI)降低(分别为0.73±0.12,0.43±0.05,0.26±0.08),细支气管Club细胞MFI增强(分别为0.49±0.07,0.73±0.08,1.02±0.19),差异均有统计学意义(P<0.05)。新生期Club细胞占气道上皮细胞百分比(9.49%±2.38%)明显低于幼年期(15.45%±3.86%)和成年期(17.23%±4.82%),差异有统计学意义(P<0.05);同时新生期增殖性Club细胞占比(6.12%±1.89%)明显高于幼年期(2.36%±0.98%)和成年期(1.94%±0.75%),差异有统计学意义(P<0.01)。新生期肺匀浆、血清CC16水平[(64.02±12.70)ng/ml,(13.91±3.36)ng/ml]均明显低于幼年期[(89.31±5.41)ng/ml,(25.77±4.68)ng/ml]和成年期[(95.74±3.31)ng/ml,(28.02±3.99)ng/ml],差异均有统计学意义(P<0.01)。结论出生后Club细胞表达丰度不断增加的主要部位为小气道,新生期是气道Club细胞增殖及分泌CC16的关键时期。Objective To clear the developing characteristics of airway Club cells in mice model.Methods The lung tissues and serum were collected from neonatal(1-week-old),infant(3-week-old)and adult(6-week-old)BALB/c mice.The immunofluorescence method was performed to detect the distribution and expression abundance of Club cells in main bronchi and bronchioles.The flow cytometry was used to determine the quantity proportion and proliferation ability,and ELISA was used to detect the levels of Club cell secreted protein(CC16)in lung homogenate and serum.Results The Club cells unevenly distributed in main bronchus of all ages of mice,while was relatively flat and uniform in bronchioles in BALB/c mice model.With increasing of mice age,the mean fluorescence intensity(MFI)of Club cells decreased in main bronchus accompanied by an increase in bronchioles with significant difference(main bronchus:0.73±0.12,0.43±0.05,0.26±0.08,respectively,P<0.05;bronchioles:0.49±0.07,0.73±0.08,1.02±0.19,respectively,P<0.05).The proportion of Club cells in airway epithelium was significantly lower in neonatal(9.49%±2.38%)than those in infant(15.45%±3.86%)and adult(17.23%±4.82%)(P<0.05).Meanwhile,the proliferative ability of Ki67 in Club cells was significantly higher in neonatal(6.12%±1.89%)than those in infant(2.36%±0.98%)and adult(1.94%±0.75%)(P<0.01).The CC16 levels in lung homogenates and serum had a statistically significant increase in infant[(89.31±5.41)ng/ml,(25.77±4.68)ng/ml]and adult[(95.74±3.31)ng/ml,(28.02±3.99)ng/ml]than those in neonatal[(64.02±12.70)ng/ml,(13.91±3.36)ng/ml](P<0.01).Conclusion Small airway is the prominent area for Club cells developing after birth,and neonatal is the critical period for Club cells proliferating and secreting CC16.

关 键 词：气道上皮 Club细胞 Club细胞分泌蛋白 BALB/C小鼠 

分 类 号：R725.6[医药卫生—儿科]