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通过定制的cDNA微阵列测定怀孕和哺乳期大鼠中与肠道离子和营养吸收相关转录本的表达。

Expression of transcripts related to intestinal ion and nutrient absorption in pregnant and lactating rats as determined by custom-designed cDNA microarray.

作者信息

Teerapornpuntakit Jarinthorn, Klanchui Amornpan, Karoonuthaisiri Nitsara, Wongdee Kannikar, Charoenphandhu Narattaphol

机构信息

Department of Physiology, Faculty of Science, Mahidol University, Rama VI Road, Bangkok, 10400, Thailand.

出版信息

Mol Cell Biochem. 2014 Jun;391(1-2):103-16. doi: 10.1007/s11010-014-1992-8. Epub 2014 Feb 12.

Abstract

In pregnancy and lactation, maternal adaptation for the enhancement of intestinal ion and nutrient absorption is of paramount importance for fetal development and lactogenesis. This nutrient hyperabsorption has been reported to result from upregulation of transporter gene expression, in part, under control of lactogenic hormone prolactin (PRL). Since a number of gene families are responsible for ion and nutrient transport in the rat small intestine, we herein developed a custom-designed cDNA microarray (CalGeneArray) to determine the transcriptome responses of duodenal epithelial cells during these reproductive periods, which was subsequently validated by quantitative real-time PCR. We thus designed 277 oligonucleotide probes to detect 113 transcripts related to ion/nutrient transport, bone/calcium metabolism, paracrine regulator, and cell metabolism. Pregnancy was found to upregulate the expressions of several duodenal transporters, e.g., Trpm6, Trpm7, Glut5, and Trpv6. Pregnant rats subjected to 7-day injection of bromocriptine, an inhibitor of PRL release, showed the increased levels of some other transcripts, e.g., insulin-2 and Cyp27b1, compared to untreated pregnant rats. Bromocriptine also increased the mRNA levels of insulin-2, glucose transporter-1 (Sglt1), and Cyp27b1, while decreasing those of Fgfr2c, Atp1b2, and Cldn19 in early lactation. During late lactation, the levels of eight studied transcripts (i.e., NaPi-IIb, Cyp27b1, Cldn18, Casr, Atp1b2, Xpnpep, Pept1, and Trpm7) were altered. In conclusion, the CalGeneArray was powerful to help reveal that pregnancy and lactation modulated the expression of genes related to duodenal nutrient transport and cell metabolism. Our findings supported the physiological significance of PRL in regulating nutrient absorption during pregnancy and lactation.

摘要

在妊娠和哺乳期,母体为增强肠道离子和营养物质吸收而进行的适应性变化对于胎儿发育和泌乳至关重要。据报道,这种营养物质的过度吸收部分是由于转运蛋白基因表达上调,这一过程受泌乳激素催乳素(PRL)的调控。由于多个基因家族负责大鼠小肠中的离子和营养物质转运,我们在此开发了一种定制的cDNA微阵列(CalGeneArray),以确定这些生殖时期十二指肠上皮细胞的转录组反应,随后通过定量实时PCR对其进行了验证。因此,我们设计了277个寡核苷酸探针来检测113种与离子/营养物质转运、骨/钙代谢、旁分泌调节因子和细胞代谢相关的转录本。研究发现,妊娠会上调几种十二指肠转运蛋白的表达,例如瞬时受体电位阳离子通道亚家族M成员6(Trpm6)、瞬时受体电位阳离子通道亚家族M成员7(Trpm7)、葡萄糖转运蛋白5(Glut5)和瞬时受体电位香草酸亚型6(Trpv6)。与未处理的妊娠大鼠相比,接受7天溴隐亭注射(一种PRL释放抑制剂)的妊娠大鼠显示出其他一些转录本水平升高,例如胰岛素-2和细胞色素P450 27B1(Cyp27b1)。溴隐亭在哺乳早期还增加了胰岛素-2、葡萄糖转运蛋白1(Sglt1)和Cyp27b1的mRNA水平,同时降低了成纤维细胞生长因子受体2C(Fgfr2c)、ATP酶1B2(Atp1b2)和紧密连接蛋白19(Cldn19)的mRNA水平。在哺乳后期,8种研究转录本(即钠-磷协同转运蛋白IIb(NaPi-IIb)、Cyp27b1、紧密连接蛋白18(Cldn18)、钙敏感受体(Casr)、Atp1b2、氨肽酶N(Xpnpep)、肽转运蛋白1(Pept1)和Trpm7)的水平发生了改变。总之,CalGeneArray有助于揭示妊娠和哺乳期调节十二指肠营养物质转运和细胞代谢相关基因的表达。我们的研究结果支持了PRL在调节妊娠和哺乳期营养物质吸收方面的生理意义。

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