Background

The overexpression of TSLP and DNA methylation in asthma were both risk factors the relationship was not clear.

Objective

This study aimed to investigate the relationship between methylation status of TSLP promoter and mRNA/protein expression in asthmatic airway epithelial cells.

Methods

Human bronchial epithelial cells were cultured in vitro and divided into: Control group, treated with PBS, model group, sensitized with LPS (10 μg/mL) for 12 h (37 °C, 5% CO₂). Other groups were cultured with the pCMV3 plasmid (M + NC/pCMV), pGPH1 plasmid (M + NC/pGPH), DNMT1/pCMV3 plasmid (M + DNMT1/pCMV), and DNMT1/pGPH1 plasmid (M + DNMT1/pGPH) for 48 h. The expression of DNA methyltransferase 1 and TSLP were measured using real-time PCR and western blotting.

Results

Compared with the control group, TSLP mRNA (1.00 ± 0.00 vs. 2.82 ± 0.81 vs. 1, P < 0.001) and protein (1.07 ± 0.04 vs. 1.46 ± 0.11, P < 0.01) were significantly greater, and the methylation of promoter was lower (92.75 ± 1.26 vs. 58.57 ± 3.34, P < 0.05) in the model group. Compared with the model group, TSLP mRNA (2.82 ± 0.81 vs. 1.17 ± 0.10, P < 0.001) decreased, but TSLP promoter methylation increased (58.57 ± 3.34 vs. 92.58 ± 7.30, P < 0.05) in M + DNMT1/pCMV. TSLP mRNA and protein were higher (2.82 ± 0.81 vs. 5.32 ± 0.21, P < 0.001; 1.46 ± 0.11 vs. 1.94 ± 0.11, respectively, P < 0.01), TSLP promoter methylation was lower (58.57 ± 3.34 vs. 33.57 ± 4.29, P < 0.05) in M + DNMT1/pGPH.

Conclusions

Overexpression of TSLP in asthmatic airway epithelial cells may be regulated by DNA demethylation.

中文翻译：

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哮喘气道上皮细胞DNA甲基化与胸腺基质淋巴细胞生成素表达的相关性

背景

哮喘中TSLP的过表达和DNA甲基化均为相关因素尚不清楚。

目的

本研究旨在探讨哮喘气道上皮细胞中TSLP启动子甲基化状态与mRNA /蛋白质表达之间的关系。

方法

在体外培养人支气管上皮细胞，分为：对照组，PBS处理，模型组，用LPS（10μg/ mL）敏化12 h（37°C，5％CO ₂）。其他组用pCMV3质粒（M + NC / pCMV），pGPH1质粒（M + NC / pGPH），DNMT1 / pCMV3质粒（M + DNMT1 / pCMV）和DNMT1 / pGPH1质粒（M + DNMT1 / pGPH）培养持续48小时。使用实时PCR和蛋白质印迹法检测DNA甲基转移酶1和TSLP的表达。

结果

与对照组相比，TSLP mRNA（1.00±0.00 vs. 2.82±0.81 vs. 1，P <0.001）和蛋白质（1.07±0.04 vs. 1.46±0.11，P <0.01）显着增加，且启动子甲基化在模型组中较低（92.75±1.26与58.57±3.34，P <0.05）。与模型组相比，在M + DNMT1 / pCMV中，TSLP mRNA（2.82±0.81 vs. 1.17±0.10，P <0.001）降低，但TSLP启动子甲基化增加（58.57±3.34 vs. 92.58±7.30，P <0.05）。TSLP mRNA和蛋白较高（2.82±0.81 vs. 5.32±0.21，P <0.001; 1.46±0.11 vs. 1.94±0.11，P <0.01），TSLP启动子甲基化较低（58.57±3.34 vs. 33.57±4.29 ，P <0.05）在M + DNMT1 / pGPH中。

结论

TSLP在哮喘气道上皮细胞中的过度表达可能受DNA脱甲基作用的调节。