p300 mediates the histone acetylation of ORMDL3 to affect airway inflammation and remodeling in asthma
p300 mediates the histone acetylation of ORMDL3 to affect airway inflammation and remodeling in asthma
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p300 介导 ORMDL3 的组蛋白乙酰化影响哮喘气道炎症和重塑
DOI：
10.1016/j.intimp.2019.105885
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发表时间：
2019-11-01
影响因子：
5.6
通讯作者：
Zhou, Qianlan
中科院分区：
文献类型：
--
作者：
Cheng, Qi;Shang, Yunxiao;Zhou, Qianlan
背景：支气管哮喘受环境和遗传因素的影响。类粘蛋白1样蛋白3（ORMDL 3）基因与儿童哮喘相关，并参与气道炎症和气道重塑。ORMDL 3启动子含有组蛋白乙酰化酶p300的结合位点。基因表达可受表观遗传修饰的影响。本研究旨在探讨p300介导的ORMDL 3基因组蛋白乙酰化（histone acetylation，HAT）对哮喘气道炎症和重塑的影响。双荧光素酶报告基因测定用于检查p300介导的HAT对ORMDL 3启动子的影响。将30只BALB/c小鼠随机分为对照组、卵清蛋白（OVA）诱导的哮喘组和哮喘+ C646（p300选择性抑制剂）组。采用无创肺功能检测各组气道高反应性（AHR）。HE染色和Masson三色染色观察气道重塑和炎症反应。采用免疫组化、western blotting和real-time PCR方法检测ORMDL 3在肺组织中的表达。ELISA和Western blotting检测肺组织HAT状态。结果：p300激活了ORMDL 3启动子的转录，导致内源性ORMDL 3 mRNA水平的增加。当p300的HAT活性丧失时，ORMDL 3启动子活性降低。哮喘小鼠肺组织中ORMDL 3表达增强，HAT活性增高。p300和aceH 3与ORMDL 3的启动子区结合。在哮喘组中，招募到ORMDL 3启动子的p300和aceH 3的量增加。结论：p300介导的HAT可调节哮喘易感基因ORMDL 3的表达，从而改善哮喘气道炎症和重塑过程。
Background: Bronchial asthma is affected by both environmental and genetic factors. The orosomucoid 1-like protein 3 (ORMDL3) gene is related to childhood asthma and is involved in airway inflammation and airway remodeling. The ORMDL3 promoter contains binding sites for the histone acetylase p300. Gene expression can be affected by epigenetic modifications. This study aimed to investigate whether the p300-mediated histone acetylation (HAT) of ORMDL3 gene affects airway inflammation and remodeling in asthma.Methods: 16HBE14o- cells were transfected with various concentrations of a wild-type p300 plasmid or p300HAT-deletion plasmids. A dual luciferase reporter assay was used to examine the effect of p300-mediated HAT on the ORMDL3 promoter. Thirty BALB/c mice were randomly divided into a control group, an ovalbumin (OVA)-induced asthma group and an asthma + C646 (a selective inhibitor of p300) group. Noninvasive lung function tests were conducted to examine airway hyperreactivity (AHR) in the different groups. HE and Masson's trichrome staining was performed to examine airway remodeling and inflammation. Immunohistochemistry, western blotting and real-time PCR were used to analyze ORMDL3 expression in lung tissues. ELISA and western blotting were used to evaluate the HAT status in lung tissue. The ChIP assay was used to determine the relationship of the ORMDL3 promoter to p300 or acetylated histone H3 (aceH3).Results: p300 activated transcription from the ORMDL3 promoter, resulting in an increase in endogenous ORMDL3 mRNA levels. ORMDL3 promoter activity was reduced when the HAT activity of p300 was lost. ORMDL3 expression was elevated, and HAT activity was high in the lung tissues of asthmatic mice. p300 and aceH3 bound to the promoter region of ORMDL3. In the asthma group, the amounts of p300 and aceH3 recruited to the ORMDL3 promoter were increased. C646 inhibited p300 expression and reduced HAT activity and aceH3 levels in asthmatic mice, thereby reducing ORMDL3 expression and relieving AHR and airway remodeling.Conclusion: p300-mediated HAT modulates the expression of the asthma susceptibility gene ORMDL3, thereby improving the process of airway inflammation and remodeling in asthma.