1 Department of Biological and Medical Sciences, Faculty of Health and Life Sciences, Oxford Brookes University , Oxford, United Kingdom.
2 Oxford Expression Technologies Ltd. , BioInnovation Hub, Oxford, United Kingdom .
Hum Gene Ther. 2017 Sep;28(9):747-756. doi: 10.1089/hum.2016.100. Epub 2016 Dec 29.
Ischemia-reperfusion (I/R) injury remains the primary contributor to delayed graft function in kidney transplantation. The beneficial application of manganese superoxide dismutase (sod), delivered by a BacMam vector, against renal I/R injury has not been evaluated previously. Therefore, this study overexpressed sod-2 in proximal tubular epithelial (HK-2) cells and porcine kidney organs during simulated renal I/R injury. Incubation of HK-2 cells with antimycin A and 2-deoxyglucose resulted in a significant decrease in intracellular adenosine triphosphate (ATP) levels; following reperfusion, ATP levels significantly increased over time in cells overexpressing sod-2. In addition, lactate dehydrogenase (LDH) release declined over 72 h in BacMam-transduced injured cells. Ex vivo delivery of sod-2 significantly increased ATP levels in organs after 24 h of cold perfusion. In vitro and ex vivo results suggested that BacMam transduction successfully delivered sod-2, which reduced injury associated with I/R, by improving ATP cell content and decreasing LDH release with a subsequent increase in kidney tissue viability. These data provide further evidence for the potential application of BacMam as a gene delivery system for attenuating injury after cold preservation.
缺血再灌注（I/R）损伤仍然是肾移植中延迟移植物功能的主要原因。以前尚未评估过通过 BacMam 载体递送锰超氧化物歧化酶（sod）对肾 I/R 损伤的有益应用。因此，本研究在模拟肾 I/R 损伤期间在近端肾小管上皮（HK-2）细胞和猪肾器官中过表达 sod-2。HK-2 细胞与安密妥因 A 和 2-脱氧葡萄糖孵育会导致细胞内三磷酸腺苷（ATP）水平显着下降；再灌注后，过表达 sod-2 的细胞中 ATP 水平随时间的推移显着增加。此外，在 BacMam 转导的受损细胞中，乳酸脱氢酶（LDH）释放量在 72 小时内下降。在冷灌注 24 小时后，sod-2 的体外递送显着增加了器官中的 ATP 水平。体外和离体结果表明，BacMam 转导成功地递送了 sod-2，通过提高 ATP 细胞含量和减少 LDH 释放，从而降低与 I/R 相关的损伤，进而增加肾脏组织活力。这些数据为 BacMam 作为减轻冷保存后损伤的基因传递系统的潜在应用提供了进一步的证据。
