Aoki Shigehisa, Noguchi Mitsuru, Takezawa Toshiaki, Ikeda Satoshi, Uchihashi Kazuyoshi, Kuroyama Hiroyuki, Chimuro Tomoyuki, Toda Shuji
Department of Pathology and Microbiology, Faculty of Medicine, Saga University, 5-1-1 Nabeshima, Saga, 849-8501, Japan.
Department of Urology, Faculty of Medicine, Saga University, 5-1-1 Nabeshima, Saga, 849-8501, Japan.
J Artif Organs. 2016 Mar;19(1):87-96. doi: 10.1007/s10047-015-0864-7. Epub 2015 Aug 30.
Peritoneal fluid dwell impacts the peritoneum by creating an abnormal physiological microenvironment. Little is known about the precise effects of fluid dwell on the peritoneum, and no adequate in vitro models to analyze the impact of fluid dwell have been established. In this study, we developed a peritoneal fluid dwell model combined with an artificial peritoneal cavity and fluid stirring generation system to clarify the effects of different dwelling solutions on the peritoneum over time. To replicate the peritoneal cavity, we devised a reconstructed peritoneal cavity utilizing a mesothelial layer, endothelial layer, and collagen membrane chamber. The reconstructed peritoneal cavity was infused with Dulbecco's modified Eagle's medium, saline, lactated Ringer's solution or peritoneal dialysis solution with repeated 4-h dwells for 10 or 20 consecutive days. The above-described solutions induced epithelial-mesenchymal transition (EMT) and hyperplasia of mesothelial cells. All solution types modulated nitric oxide synthase activities in mesothelial and endothelial cells and nitric oxide concentrations in dwelling solutions. Inhibition of nitric oxide synthase activity acted synergistically on mesothelial EMT and hyperplasia. The present findings suggest that solutions infused into the peritoneal cavity are likely to affect nitric oxide production in the peritoneum and promote peritoneal fibrosis. Our newly devised peritoneal cavity model should be a promising tool for understanding peritoneal cellular kinetics and homeostasis.
腹腔内液体潴留通过创造异常的生理微环境来影响腹膜。关于液体潴留对腹膜的确切影响知之甚少，并且尚未建立足够的体外模型来分析液体潴留的影响。在本研究中，我们开发了一种结合人工腹腔和液体搅拌生成系统的腹腔内液体潴留模型，以阐明不同潴留溶液随时间对腹膜的影响。为了复制腹腔，我们设计了一种利用间皮细胞层、内皮细胞层和胶原膜腔室构建的重建腹腔。向重建腹腔内注入杜氏改良伊格尔培养基、生理盐水、乳酸林格氏液或腹膜透析液，并重复4小时的潴留，持续10或20天。上述溶液诱导间皮细胞上皮-间质转化（EMT）和增生。所有溶液类型均调节间皮细胞和内皮细胞中的一氧化氮合酶活性以及潴留溶液中的一氧化氮浓度。一氧化氮合酶活性的抑制对间皮EMT和增生起协同作用。目前的研究结果表明，注入腹腔的溶液可能会影响腹膜中一氧化氮的产生并促进腹膜纤维化。我们新设计的腹腔模型应该是理解腹膜细胞动力学和内环境稳定的一个有前景的工具。
