摘要
目的探讨环磷腺苷效应元件结合蛋白(CREB)在3岁以下反复喘息患儿中的表达水平及其对血清类黏蛋白1样蛋白3基因(ORMDL3)表达水平的影响。方法选取2017年6月至2019年6月就诊的3岁以下反复喘息儿童36例为病例组,另选取行健康体检的24例儿童为健康对照组,使用荧光定量PCR法检测外周血CREB的表达水平。培养人支气管上皮细胞(BEAS-2B),使用双荧光素酶报告基因和荧光定量PCR法检测过表达和siRNA干扰CREB对BEAS-2B细胞ORMDL3基因启动子活性及m RNA表达水平的影响。结果病例组CREB表达水平高于健康对照组(P<0.001)。在BEAS-2B细胞中,过表达CREB能上调ORMDL3基因的启动子活性及m RNA表达水平(P<0.05);siRNA干扰CREB的表达后ORMDL3基因的启动子活性及m RNA表达水平均降低(P<0.05)。结论反复喘息患儿CREB表达增加,其可能通过调控ORMDL3基因的表达参与了反复喘息的发病。
Objective To study the expression level of cAMP response element-binding protein(CREB)in children with recurrent wheezing under three years of age and its effect on the expression of the serum orosomucoid 1-like protein 3(ORMDL3)gene.Methods Thirty-six children with recurrent wheezing under three years of age who visited the hospital from June 2017 to June 2019 were selected as the recurrent wheezing group.Twenty-four healthy children from physical examination were selected as the control group.The CREB expression level in peripheral blood was measured by quantitative real-time PCR.Human bronchial epithelial cells(BEAS-2 B)were cultured,and dualluciferase reporter assay and quantitative real-time PCR were used to investigate the effects of overexpression and siRNA interference of CREB on the promoter activity and mRNA expression of the ORMDL3 gene in the BEAS-2 B cells.Results The expression level of CREB in the recurrent wheezing group was significantly higher than that in the control group(P<0.001).In BEAS-2 B cells,overexpression of CREB significantly up-regulated the promoter activity and mRNA expression of the ORMDL3 gene(P<0.05),while siRNA interference of CREB significantly reduced the promoter activity and mRNA expression of the ORMDL3 gene(P<0.05).Conclusions The expression of CREB is increased in children with recurrent wheezing,and CREB may be involved in the pathogenesis of recurrent wheezing by regulating expression of the ORMDL3 gene.
作者
金蕊
张振兴
朱亮华
庄丽丽
陈筱青
JIN Rui;ZHANG Zhen-Xing;ZHU Liang-Hua;ZHUANG Li-Li;CHEN Xiao-Qing(Department of Pediatrics,First Affiliated Hospital,Nanjing Medical University,Nanjing 210029,China)
出处
《中国当代儿科杂志》
CAS
CSCD
北大核心
2020年第9期980-983,共4页
Chinese Journal of Contemporary Pediatrics
基金
江苏省科教强卫工程青年医学人才项目(QNRC2016615)
江苏省妇幼健康重点人才项目(FRC201736)。
