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Increased ENaC activity during kidney preservation in Wisconsin solution

机译:在威斯康星州的肾脏保存过程中增加了ENaC活性

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The invention of an effective kidney preservation solution capable of prolonging harvested kidney viability is the core of kidney transplantation procedure. Researchers have been working on upgrading the preservation solution quality aiming at prolonging storage time while maintaining utmost organ viability and functionality. For many years, the University of Wisconsin (UW) solution has been considered the gold standard solution for kidney preservation. However, the lifespan of kidney preservation in the UW solution is still limited. Its impact on the epithelial Na+ channel (ENaC) activity and its mediated processes is unknown and the primary goal of this study. Kidneys harvested from 8?weeks old Sprague Dawley rats were divided into 4 groups depending upon the period of preservation in UW solution. Additional analysis was performed using dogs’ kidneys. ENaC activity was measured using patch clamp technique; protein expression and mRNA transcription were tested through Western blot and RT-qPCR, respectively. A colorimetric LDH level estimation was performed at different time points during UW solution preservation. Kidney preservation in Wisconsin solution caused reduction of the kidney size and weight and elevation of LDH level. ENaC activity increased in both rat and dog kidneys preserved in the UW solution as assessed by patch clamp analysis. On the contrary, ENaC channel mRNA levels remained unchanged. ENaC activity is significantly elevated in the kidneys during preservation in UW solution, which might affect the immediate post-implantation allograft function and trajectory post-transplant.
机译:能够延长收获的肾脏生存力的有效肾脏保存溶液的发明是肾脏移植程序的核心。研究人员一直致力于提高保存溶液的质量,以期延长保存时间,同时保持最大的器官活力和功能。多年来,威斯康星大学(UW)解决方案一直被认为是肾脏保存的金标准溶液。但是,UW溶液中肾脏保存的寿命仍然有限。其对上皮Na +通道(ENaC)活性及其介导过程的影响尚不清楚,这是本研究的主要目标。根据在UW溶液中保存的时间,将从8周大的Sprague Dawley大鼠收获的肾脏分为4组。使用狗的肾脏进行了进一步的分析。使用膜片钳技术测量ENaC活性;通过Western blot和RT-qPCR分别检测蛋白的表达和mRNA的转录。在UW溶液保存期间的不同时间点进行了比色LDH含量估算。肾脏在威斯康星州溶液中的保存导致肾脏大小和体重的减少以及LDH水平的升高。通过膜片钳分析评估,保存在UW溶液中的大鼠和狗肾脏的ENaC活性均增加。相反,ENaC通道mRNA水平保持不变。在UW溶液中保存期间,肾脏中的ENaC活性显着升高,这可能会影响立即的同种异体移植功能和移植后的轨迹。

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