Ischaemia-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, in this study we 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 ATP levels; following reperfusion, ATP levels significantly increased overtime 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.
Hitchman, ElisabettaHitchman, Richard B.King, Linda A.
Faculty of Health and Life Sciences
Year of publication: 2017Date of RADAR deposit: 2017-01-17