咖啡因通过肺组织自噬对高氧致新生鼠支气管肺发育不良的影响研究  

Effect of caffeine on bronchopulmonary dysplasia in neonatal rats induced by hyperoxia through autophagy

作　　者：盛安群 王石 刘园园 陈翠娥 张希希 卢建双 孙媛媛[5] SHENG Anqun;WANG Shi;LIU Yuanyuan;CHEN Cui'e;ZHANG Xixi;LU Jianshuang;SUN Yuanyuan(Department of Pediatrics,the First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325000,China;不详)

机构地区：[1]温州医科大学附属第一医院儿科,325000 [2]浙江大学医学院附属妇产科医院麻醉科 [3]义乌市妇幼保健院儿科 [4]玉环市人民医院儿科 [5]温州医科大学附属衢州医院儿科

出　　处：《浙江医学》2024年第24期2582-2588,I0003,共8页Zhejiang Medical Journal

基　　金：温州市基础性科研项目(Y20190226、Y2023003);浙江省医学会临床医学科研专项资金项目(2024ZYC-B51)。

摘　　要：目的探讨咖啡因对高氧致新生大鼠支气管肺发育不良(BPD)的影响及其作用机制。方法将48只SD新生大鼠按随机数字表法分为空气组、高氧组、空气+咖啡因组、高氧+咖啡因组,每组12只。除空气组外,其余3组构建BPD模型,空气+咖啡因组和高氧+咖啡因组腹腔注射20 mg/kg咖啡因,第7、14天收集大鼠肺组织。HE染色评估4组大鼠肺组织形态学变化,免疫组化法检测肺组织CD31蛋白平均光密度(AOD)值,qRT-PCR法检测肺组织CD31、微管相关蛋白1轻链3B(LC3B)、p62 mRNA相对表达量,免疫荧光法检测肺组织LC3B、p62蛋白免疫荧光强度,Western blot法检测肺组织LC3B-Ⅱ和LC3B-Ⅰ蛋白比值(LC3B-Ⅱ/LC3B-Ⅰ)、p62蛋白相对表达量。结果出生后第7、14天,与空气组相比,高氧组大鼠肺泡辐射状计数(RAC)值显著降低,平均内衬间隔(MLI)值显著增加,肺组织CD31蛋白AOD值、CD31 mRNA相对表达量显著降低,LC3B mRNA相对表达量、LC3B蛋白免疫荧光强度、LC3B-Ⅱ/LC3B-Ⅰ显著下降,p62 mRNA相对表达量、p62蛋白免疫荧光强度、p62蛋白相对表达量显著升高,差异均有统计学意义(均P<0.05);与高氧组相比,高氧+咖啡因组大鼠肺泡RAC值显著升高、MLI值显著减少,肺组织CD31蛋白AOD值、CD31 mRNA相对表达量显著升高,LC3B mRNA相对表达量、LC3B蛋白免疫荧光强度及LC3B-Ⅱ/LC3B-Ⅰ显著升高,p62 mRNA相对表达量、p62蛋白免疫荧光强度、p62蛋白相对表达量显著下降,差异均有统计学意义(均P<0.05)。结论高氧致BPD大鼠肺组织自噬不足,咖啡因干预可改善BPD新生大鼠肺组织自噬及肺发育受阻,这为咖啡因治疗BPD提供了新的理论依据。Objective To explore the effect of caffeine on hyperoxia-induced bronchopulmonary dysplasia(BPD)in neonatal rats and related mechanism.Methods Forty-eight SD neonatal rats were randomly divided into air group,hyperoxia group,air+caffeine group,and hyperoxia+caffeine group,with 12 rats in each.The animals were exposed to hyperoxia to induce BPD model,and rats in air+caffeine and hyperoxia+caffeine groups were injected intraperitoneally with 20 mg/kg of caffeine,lung tissue samples were collected on 7th and 14th days after modelling.HE staining was used to evaluate the morphological changes in lung tissue,immunohistochemistry was used to detect the average optical density(AOD)value of CD31 protein in lung tissue,qRT-PCR was used to detect the relative expression of CD31,microtubule-associated protein 1 light chain 3B(LC3B)and p62 mRNA,immunofluorescence was used to detect the immunofluorescence intensity of LC3B and p62 protein,and Western blot was used to detect the ratio of LC3B-Ⅱ to LC3B-I protein(LC3B-Ⅱ/LC3B-Ⅰ)and the relative expression of p62 protein in lung tissue.Results On the 7th and 14th days after birth,compared with the air group,the value of radial alveolar count(RAC)was significantly decreased,the mean linear intercept(MLI)was significantly increased,the the AOD value of CD31 protein and the relative expression of CD31 mRNA in lung tissue were significantly decreased in the hyperoxia group;The relative expression of LC3B mRNA,the immunofluorescence intensity of LC3B protein,and the LC3B-Ⅱ/LC3B-Ⅰ in lung tissue were significantly decreased,while the relative expression of p62 mRNA,the immunofluo-rescence intensity of p62 protein,and the relative expression of p62 protein were significantly increased(all P<0.05).Compared with the hyperoxia group,the hyperoxia+caffeine group showed a significant increase in RAC value and a decreased in MLI value,a significant increase in the AOD value of CD31 protein and the relative expression of CD31 mRNA,the relative expression of LC3B mRNA,the immunofluore

关 键 词：咖啡因 自噬 支气管肺发育不良 高氧 

分 类 号：R722.1[医药卫生—儿科]