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首页> 外文期刊>World Journal of Gastroenterology >Hormonal regulation of dipeptide transporter (PepT1) in Caco-2 cells with normal and anoxia/reoxygenation management
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Hormonal regulation of dipeptide transporter (PepT1) in Caco-2 cells with normal and anoxia/reoxygenation management

机译：具有正常和缺氧/复氧管理的Caco-2细胞中二肽转运蛋白（PepT1）的激素调节

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AIM: To determine the regulation effects of recombinant human growth hormone (rhGH) on dipeptide transporter (PepT1) in Caco-2 cells with normal culture and anoxia/ reoxygenation injury. METHODS: A human intestinal cell monolayer (Caco-2) was used as the in vitro model of human small intestine and cephalexin as the model substrate for dipeptide transporter (PepT1). Caco-2 cells grown on Transwell membrane filters were preincubated in the presence of rhGH in the culture medium for 4 d, serum was withdrawn from monolayers for 24 h before each experiment. The transport experiments of cephalexin across apical membromes were then conducted; Caco-2 cells grown on multiple well dishes (24 pore) with normal culture or anoxia/reoxygenation injury were preincubated with rhGH as above and uptake of cephalexin was then measured. RESULTS: The transport and uptake of cephelaxin across apical membranes of Caco-2 cells after preincubation with rhGH were significantly increased compared with controls (P=0.045, 0.0223). Also, addition of rhGH at physiological concentration (34 nM) to incubation medium greatly stimulates cephalexin uptake by anoxia/reoxygenation injuried Caco-2 cells (P=0.0116), while the biological functions of PepT1 in injured Caco-2 cells without rhGH were markedly downregulated. Northern blot analysis showed that the level of PepT1 mRMA of rhGH-treated injured Caco-2 cells was greatly increased compared to controls. CONCLUSION: The present results of rhGH stimulating the uptake and transport of cephalexin indicated that rhGH greatly upregulates the physiological effects of dipeptide transporters of Caco-2 cells. The alteration in the gene expression may be a mechanism of regulation of PepT1. In addition, Caco-2 cells take up cephalexin by the Proton-dependent dipeptide transporters that closely resembles the transporters present in the intestine. Caco-2 cells represent an ideal cellular model for future studies of the dipeptide transporter.
机译：目的：确定重组人生长激素（rhGH）对正常培养和缺氧/复氧损伤的Caco-2细胞中二肽转运蛋白（PepT1）的调节作用。方法：以人小肠单层细胞（Caco-2）为模型，以头孢氨苄为二肽转运蛋白（PepT1）的体外模型。将在Transwell膜滤器上生长的Caco-2细胞在存在rhGH的条件下在培养基中预孵育4 d，在每次实验前从单层中提取血清24 h。然后进行了头孢氨苄跨过顶端膜的转运实验。如上所述，将在正常培养或缺氧/复氧损伤的多个孔皿（24孔）上生长的Caco-2细胞与rhGH一起预孵育，然后测量头孢氨苄的摄取。结果：与对照组相比，经rhGH预孵育后，头孢氨苄跨Caco-2细胞顶膜的转运和摄取显着增加（P = 0.045，0.0223）。此外，在培养液中添加生理浓度（34 nM）的rhGH可以极大地刺激缺氧/复氧损伤的Caco-2细胞对头孢氨苄的摄取（P = 0.0116），而在没有rhGH的受损Caco-2细胞中PepT1的生物学功能显着下调。 Northern印迹分析表明，与对照相比，rhGH处理的受伤的Caco-2细胞的PepT1 mRMA水平大大提高。结论：rhGH刺激头孢氨苄的摄取和转运的最新结果表明，rhGH极大地上调了Caco-2细胞二肽转运蛋白的生理作用。基因表达的改变可能是调控PepT1的机制。此外，Caco-2细胞通过质子依赖性二肽转运蛋白摄取头孢氨苄，该转运蛋白与肠道中存在的转运蛋白极为相似。 Caco-2细胞代表了二肽转运蛋白未来研究的理想细胞模型。

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