CREB在反复喘息患儿中的表达及对ORMDL3基因表达的影响

Expression of CREB in children with recurrent wheezing and its effect on ORMDL3 gene expression

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mRNA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mRNA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mRNA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。

Abstract:

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-2B) were cultured, and dual-luciferase 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-2B 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-2B 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.