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目的探索重组法国梧桐花粉过敏原2(rPla a2)对小鼠NIH3T3细胞血清类黏蛋白1样蛋白3(ORMDL3)表达的影响及相关调控机制。方法实时定量PCR和Western blot法分别检测rPla a2对NIH3T3细胞ORMDL3和GATA3 mRNA和蛋白水平的影响;通过干扰和过表达观察GATA结合蛋白3(GATA3)在rPla a2调节ORMDL3表达中的作用;通过双荧光素酶活性分析检测rPla a2和GATA3对ORMDL3基因启动子活性的影响。结果 rPla a2增强NIH3T3细胞ORMDL3基因启动子活性,并增加其mRNA水平和蛋白表达;同时增加GATA3 mRNA和蛋白表达。敲低GATA3抑制rPla a2对ORMDL3基因的诱导作用,过表达GATA3则可增强该诱导作用。结论 GATA3可介导rPla a2对NIH3T3细胞ORMDL3基因的转录调控。
Objective To explore the effect of rPla a2 on the expression of serum mucin-like protein-3 (ORMDL3) in mouse NIH3T3 cells and its related regulatory mechanism. Methods The effects of rPla a2 on ORMDL3 and GATA3 mRNA and protein levels in NIH3T3 cells were detected by real-time PCR and Western blot respectively. The effect of GATA binding protein 3 (GATA3) on the regulation of ORMDL3 expression was observed by interference and overexpression. The luciferase activity assay was used to detect the effect of rPla a2 and GATA3 on the promoter activity of ORMDL3 gene. Results rPla a2 enhanced the promoter activity of ORMDL3 gene in NIH3T3 cells and increased the mRNA and protein expression of GDNF3 and increased the expression of GATA3 mRNA and protein. Knockdown of GATA3 inhibited rPla a2 induction of ORMDL3 gene, and overexpression of GATA3 enhanced this induction. Conclusion GATA3 can mediate the transcriptional regulation of ORMDL3 gene by NIH3T3 cells by rPla a2.