Department of Nephrology and Rheumatology, Children's Hospital of Fudan University, No. 399 Wanyuan Road, Shanghai, 201102, China.
Shanghai Kidney Development and Pediatric Kidney Disease Research Center, Shanghai, China.
BMC Mol Biol. 2018 Mar 20;19(1):4. doi: 10.1186/s12867-018-0104-9.
Bicarbonate-based peritoneal dialysis (PD) fluids enhance the migratory capacity and damage-repair ability of human peritoneal mesothelial cells by upregulating AQP1. However, little is known about the underlying molecular mechanisms.
Here we used HEK-293T cells to investigate the effect of pH on AQP1 gene transcription levels. We found that AQP1 mRNA levels increases with pH. Transfection of HEK-293T cells with luciferase reporter vectors containing different regions of the AQP1 promoter identified an upstream region in the AQP1 gene between - 2200 and - 2300 bp as an enhancer required for pH-mediated regulation of AQP1 expression. Site-directed mutagenesis of this specific promoter region revealed a critical region between - 2257 and - 2251 bp, and gene knock-down experiments and ChIP assays suggested that the Spi-B transcription factor SPIB is involved in pH-mediated regulation of AQP1 expression.
We identified an upstream region in the AQP1 gene and the transcription factor SPIB that are critically involved in pH-mediated regulation of AQP1 expression. These findings provide the basis for further studies on the pH- and buffer-dependent effects of PD fluids on peritoneal membrane integrity and function.
碳酸氢盐腹膜透析（PD）液通过上调水通道蛋白 1（AQP1）来增强人腹膜间皮细胞的迁移能力和损伤修复能力。然而，其潜在的分子机制知之甚少。
本研究使用 HEK-293T 细胞来研究 pH 对 AQP1 基因转录水平的影响。结果发现 AQP1 mRNA 水平随 pH 值的升高而增加。通过转染包含 AQP1 启动子不同区域的荧光素酶报告载体，我们确定了 AQP1 基因中位于-2200 至-2300bp 之间的上游区域是 pH 介导的 AQP1 表达调控所必需的增强子。对该特定启动子区域的定点突变揭示了一个关键区域，位于-2257 至-2251bp 之间，基因敲低实验和 ChIP 分析表明，SPIB 转录因子 SPIB 参与了 pH 介导的 AQP1 表达调控。
我们确定了 AQP1 基因中的一个上游区域和转录因子 SPIB，它们在 pH 介导的 AQP1 表达调控中起着关键作用。这些发现为进一步研究 PD 液的 pH 值和缓冲依赖性对腹膜膜完整性和功能的影响提供了基础。
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