丙戊酸钠对哮喘相关基因ORMDL3的作用及其机制研究
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Effect of sodium valprate on ORMDL3 transcriptional regulation and its possible mechanism
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    摘要:

    目的:探讨丙戊酸钠对人血清类黏蛋白1样蛋白3(ORMDL3)转录表达的影响及其机制。方法:利用RT-PCR测定丙戊酸钠对人ORMDL3 mRNA表达的影响;通过点突变、双光荧素酶报告基因分析方法确定丙戊酸钠对ORMDL3启动子活性的影响及其作用机制。结果:在人胚肾293细胞(HEK293)中,丙戊酸钠(终浓度2 mmol/L)可明显降低哮喘ORMDL3基因的表达量。荧光素酶分析结果显示:丙戊酸钠能够明显抑制ORMDL3基因的启动子活性;对人ORMDL3核心启动子区正向调控ORMDL3表达的p300、CREB位点逐个突变后,丙戊酸钠对突变质粒的降低作用较野生型质粒无显著性差异。结论:丙戊酸钠可下调HEK293细胞ORMDL3转录表达水平,此调节作用并非通过降低与ORMDL3核心启动子区p300和CREB位点的结合而发挥。

    Abstract:

    Objective: The main purpose is to investigate the effect and mechanism of sodium valprate on the transcriptional expression of the human orosomucoid 1-like protein3(ORMDL3). Methods: The effect of sodium valprate on the mRNA expression of ORMDL3 was determined by using semi-quantitative RT-PCR. The effect of sodium valprate on ORMDL3 promoter activity and its mechanism were detected by point mutations and Dual-Luciferase reporter assays. Results: By adding sodium valprate(final concentration of 2 mmol/L) in human embryonic kidney 293 cells (HEK293 cells), it can significantly reduce the expression amount of the human ORMDL3 gene. The luciferase analysis showed that sodium valprate can significantly inhibit the promoter activity of ORMDL3 gene. P300 and CREB binding to the promoter region can drive the ORMDL3 transcription. When sodium valprate act on the plasmids without the binding region of p300 or CREB, there was no significant difference compared with the wild-type plasmid. Conclusion: Sodium valprate can reduce the mRNA level of the human ORMDL3 in HEK293 cells independently of binding with the p300 and CREB in core promoter area of ORMDL3 gene.

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章素芬,金 蕊,周国平.丙戊酸钠对哮喘相关基因ORMDL3的作用及其机制研究[J].南京医科大学学报(自然科学版),2017,(12):1587-1590

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  • 收稿日期:2016-12-07
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  • 在线发布日期: 2017-12-29
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