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首页> 外文期刊>American Journal of Physiology >Transcriptional inhibition of intestinal NHE8 expression by glucocorticoids involves Pax5.
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Transcriptional inhibition of intestinal NHE8 expression by glucocorticoids involves Pax5.

机译：糖皮质激素对肠道NHE8表达的转录抑制作用涉及Pax5。

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摘要

Sodium/hydrogen exchangers (NHEs) are a family of proteins that transport sodium ions into the cells by moving protons out of the cells. They play a major role in sodium absorption, cell volume regulation, and intracellular pH regulation. Three out of nine identified NHEs (NHE2, NHE3, and NHE8) are expressed on the apical membrane of intestinal epithelial cells. Glucocorticoids have been found to regulate NHE3 function in the intestine, but it is unknown if they have a similar function on NHE8 expression. Interestingly, high glucocorticoid levels in the intestine coincide chronologically with the change from high expression of NHE8 to high expression of NHE3. Studies were performed to explore the role of glucocorticoids on NHE8 expression during intestinal maturation. Brush-border membrane vesicles were isolated from intestinal epithelia, and Western blotting was performed to determine NHE8 protein expression of suckling male rats treated with methylpredisolone. Real-time PCR was used to quantitate NHE8 mRNA expression in rats and Caco-2 cells. Human NHE8 promoter activity was characterized through transfection of Caco-2 cells. Gel mobility shift assays (GMSAs) were used to identify the promoter sequences and the transcription factors involved in glucocorticoid-mediated regulation. Our results showed that the expression of NHE8 mRNA and protein was decreased in glucocorticoid-treated rats and human intestinal epithelial cells (Caco-2). The activity of the human NHE8 gene promoter transfected in Caco-2 cells was also reduced by glucocorticoid treatment. GMSAs suggested that the reduction in promoter activity in the presence of glucocorticoids was due to enhanced transcription factor Pax5 binding on the NHE8 proximal promoter region. In conclusion, this study showed that glucocorticoids inhibit NHE8 gene expression by increasing Pax5 binding on NHE8 gene promoter, suggesting an important role for Pax5 during intestinal maturation.
机译：钠/氢交换剂（NHE）是一类蛋白质，它们通过将质子移出细胞，将钠离子转运到细胞中。它们在钠吸收，细胞体积调节和细胞内pH调节中起主要作用。九个已鉴定的NHE中的三个（NHE2，NHE3和NHE8）在肠上皮细胞的顶膜上表达。已经发现糖皮质激素可调节肠道中的NHE3功能，但尚不清楚它们是否对NHE8表达具有类似的功能。有趣的是，肠道中高糖皮质激素水平与从NHE8的高表达到NHE3的高表达的时间顺序一致。进行研究以探索糖皮质激素在肠成熟过程中对NHE8表达的作用。从肠上皮中分离刷状边界膜囊泡，并进行蛋白质印迹法测定用甲基强的松龙治疗的乳鼠雄性大鼠的NHE8蛋白表达。实时荧光定量PCR用于定量大鼠和Caco-2细胞中NHE8 mRNA的表达。人NHE8启动子活性通过转染Caco-2细胞来表征。凝胶迁移率迁移分析（GMSA）用于鉴定糖皮质激素介导的调控中涉及的启动子序列和转录因子。我们的结果表明，在糖皮质激素治疗的大鼠和人肠上皮细胞（Caco-2）中，NHE8 mRNA和蛋白的表达降低。糖皮质激素处理还降低了在Caco-2细胞中转染的人类NHE8基因启动子的活性。 GMSA提示，在糖皮质激素存在下启动子活性降低是由于NHE8近端启动子区域上转录因子Pax5结合增强。总之，这项研究表明糖皮质激素通过增加NHE8基因启动子上的Pax5结合来抑制NHE8基因表达，表明Pax5在肠道成熟过程中起着重要作用。

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