Attenuating ischemia/reperfusion injury in rat cardiac transplantation by intracoronary infusion with siRNA cocktail solution.

Bo Yang,Jin Wang,Lai Wei,Bo Zhang,Meixi Wang,Zhishui Chen,Dong Chen,Chen Dai,Wu Duan,Zhenyi Han,Yuanyuan Zhao

doi:10.1042/bsr20193937

Abstract

Tumor necrosis factor-α (TNF-α), caspase-8, and complement component 5a receptor (C5aR) are known to play a crucial role in the myocardial ischemia/reperfusion (I/R) injury in cardiac transplantation. We hypothesized that the intracoronary infusion of TNF-α, caspase-8, and C5aR small interfering RNAs (siRNA) would protect cardiac allograft function and improve graft survival from I/R injury-induced organ failure. I/R injury of cardiac allograft was induced by syngeneic rat cardiac transplantation, in which the transplanted hearts were infused with saline or different amounts of siRNA cocktail solution targeting TNF-α, caspase-8, and C5aR via coronary arteries, and subsequently subjected to 18 h of preservation at 4°C in histidine–tryptophan–ketoglutarate (HTK) solution. The effects of siRNA cocktail solution on prolonged cold I/R injury were determined by assessing graft survival, histopathological changes, myeloperoxidase (MPO) activity, and malondialdehyde (MDA) concentration. The perfused siRNA cocktail solution successfully knocked down the expression of TNF-α, caspase-8, and C5aR in vitro and in vivo. Approximately 91.7% of control hearts that underwent 18 h of cold ischemia ceased their function after transplantation; however, 87.5% of cardiac allografts from the highest dose siRNA cocktail solution-pretreated hearts survived >14 days and exhibited minimal histological changes, with minimal cellular infiltration, interstitial edema, and inflammation and maximal reduced MPO activity and MDA concentration in the cardiac allograft. We demonstrated the feasibility and efficiency of infusion of TNF-α, caspase-8, and C5aR siRNA via the intracoronary route as a promising strategy for gene silencing against I/R injury in cardiac transplantation.

Highlights

Organ transplantation is a common therapy for end-stage organ failure
Before the small interfering RNAs (siRNA) intracoronary delivery to rat heart, we examined whether siRNA could suppress Tumor necrosis factor-α (TNF-α), caspase-8, or component 5a receptor (C5aR) expression efficiently
We have demonstrated that intracoronary infusion of siRNA cocktail solution can achieve TNF-α, caspase-8, and C5aR gene silence in rat cardiac allografts and result in myocardial protection after I/R injury induced by cold preservation

Summary

Introduction

Organ transplantation is a common therapy for end-stage organ failure. Transplantation requires cold storage of organs and warm blood transported back to them. Ischemia/reperfusion (I/R) injury caused by this process is still a serious problem affecting the outcome of transplantation [1]. I/R injury usually results in early graft dysfunction or primary nonfunction. I/R injury plays an important role in the development of chronic graft failure [2,3]. I/R injury involves a series of cascade reactions, such as hypoxia, acidosis, and tissue damage, which lead to many post-transplantation events, including cytokines, chemokines, up-regulation of complement, and cell apoptosis [4,5,6]

Methods

Results

Conclusion