权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Regulation of AQP expression by serotonin and pathophisiology of IBS

血清素对 AQP 表达的调节和 IBS 的病理生理学

基本信息

批准号：
15590641
负责人：
TSUJIKAWA Tomoyuki
金额：
$ 2.24万
依托单位：
Shiga University of Medical Science
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590641/
关键词：
IBS serotonin AQP3 serotonin receptor natriuretic peptides 5-HT3 receptor アクアポリン過敏性腸症候群

项目摘要

Some serotonin receptor antagonists have been used as a therapy for irritable bowel syndrome. The mechanism of this action was thought to control intestinal movement via down regulation of serotonin in the intestinal neuron. We reported that the expression of AQP3 in cultured human colonic epithelial cells HT-29 was upregulated by vasoactive intestinal polypeptide. Although Ht-29 cells express 5-HT2a,5-HT3 and 5-HT4 receptors, the addition of serotonin did not alter the expression of AQP3 mRNA. To characterize the regulation of AQP3 expression by atrial natriuretic peptide(ANP) and brain natriuretic peptide(BNP), we studied mRNA expression by Northern blotting, protein expression by Western blotting and DNA binding activity by electrophoretic mobility shift assay(EMSA) in HT-29. We also used several inhibitors to investigate signal transduction. AQP3 mRNA was up-regulated in addition to ANP (more than 100 nM) and BNP (more than 10 nM). The expression of AQP3 protein was enhanced at 1 hour after the addition of ANP and BNP. The combination of protein kinase-A(PK-A) and protein kinase-G(PK-G) inhibitors completely inhibited the expression of AQP3 mRNA enhanced by ANP or BNP to its basal level. The EMSA of the cyclic-AMP response element(CRE) in HT-29 cells revealed a single band. These results indicate that ANP and BNP up-regulated the expression of AQP3 mRNA and protein, and both PK-A and PK-G dependent pathways mediated this effect.

一些血清素受体拮抗剂已被用作肠易激综合征的治疗方法。这种作用的机制被认为是通过下调肠道神经元中的血清素来控制肠道运动。我们报道了培养的人结肠上皮细胞HT-29中AQP3的表达被血管活性肠多肽上调。尽管Ht-29细胞表达5-HT2a、5-HT3和5-HT4受体，但添加5-羟色胺并没有改变AQP3 mRNA的表达。为了表征心房钠尿肽（ANP）和脑钠尿肽（BNP）对AQP3表达的调节，我们通过RNA印迹法研究了HT-29中的mRNA表达，通过蛋白质印迹法研究了蛋白质表达，并通过电泳迁移率变动分析（EMSA）研究了DNA结合活性。我们还使用了几种抑制剂来研究信号转导。除了 ANP（超过 100 nM）和 BNP（超过 10 nM）外，AQP3 mRNA 也上调。添加ANP和BNP后1小时AQP3蛋白表达增强。蛋白激酶-A（PK-A）和蛋白激酶-G（PK-G）抑制剂联合应用可将ANP或BNP增强的AQP3 mRNA表达完全抑制至基础水平。 HT-29 细胞中环磷酸腺苷反应元件 (CRE) 的 EMSA 显示单条带。这些结果表明，ANP 和 BNP 上调 AQP3 mRNA 和蛋白的表达，并且 PK-A 和 PK-G 依赖性途径介导了这种作用。