Chen Jian-An, Chang Li-Ren, Feng Guan-Ming, Lee Shu-Ting, Hsieh Chien-Yan, Jeng Seng-Feng, Huang Wen-Shyan
Division of Plastic Surgery, Kaohsiung Armed Forces General Hospital, Kaohsiung 802, Taiwan, R.O.C. ; Department of Plastic Surgery, E-DA Hospital, Kaohsiung 824, Taiwan, R.O.C.
Department of Plastic Surgery, E-DA Hospital, Kaohsiung 824, Taiwan, R.O.C.
Exp Ther Med. 2015 Nov;10(5):1967-1972. doi: 10.3892/etm.2015.2771. Epub 2015 Sep 23.
Aquaporins (AQPs) are widely-expressed small water channel proteins that provide the major route for water transport across plasma membranes in various cell types. Although the quantity of water transported in the intestinal tract is second only to that in the kidney, the precise role of AQPs in this organ remains largely uncertain. The present study reports the effects of hypertonic stress and ischemia/reperfusion injury on the expression of AQPs in intestinal epithelial cells. Cultured rat intestinal epithelial cells were incubated in 300 mM mannitol-containing, hypertonic culture medium or subjected to simulated ischemia/reperfusion treatment. The cell viability was evaluated by MTT assay, and the expression of AQPs was determined by semi-quantitative reverse transcription polymerase chain reaction and western blotting. Despite reduced viability, the cells exposed to hypertonic stress for 16 h demonstrated enhanced expression of AQP1 mRNA and protein. AQP9 and glycosylated AQP11 proteins were also markedly upregulated. Ischemia alone did not affect the cell viability, but subsequent reperfusion significantly reduced viability. The mRNA expression levels of all the tested AQPs were not altered by ischemia alone or by ischemia/reperfusion; however, AQP8 protein was markedly reduced by ischemic injury. In addition, treatment with ischemia alone eradicated the normally-expressed, non-glycosylated AQP11 protein whilst inducing pronounced expression of the glycosylated form. These observations may indicate that AQPs function in the intestinal epithelia in response to stress.
水通道蛋白(AQPs)是广泛表达的小水通道蛋白,为各种细胞类型中水分子跨质膜运输提供了主要途径。尽管肠道中运输的水量仅次于肾脏,但AQPs在该器官中的具体作用仍很大程度上不确定。本研究报告了高渗应激和缺血/再灌注损伤对肠上皮细胞中AQPs表达的影响。将培养的大鼠肠上皮细胞置于含300 mM甘露醇的高渗培养基中孵育或进行模拟缺血/再灌注处理。通过MTT法评估细胞活力,并通过半定量逆转录聚合酶链反应和蛋白质印迹法测定AQPs的表达。尽管细胞活力降低,但暴露于高渗应激16小时的细胞显示AQP1 mRNA和蛋白表达增强。AQP9和糖基化的AQP11蛋白也显著上调。单独缺血不影响细胞活力,但随后的再灌注显著降低了活力。单独缺血或缺血/再灌注均未改变所有测试AQPs的mRNA表达水平;然而,缺血损伤使AQP8蛋白显著减少。此外,单独缺血处理消除了正常表达的非糖基化AQP11蛋白,同时诱导了糖基化形式的明显表达。这些观察结果可能表明AQPs在肠上皮细胞中对应激起作用。