Normothermic Machine Perfusion Combined with Bone Marrow Mesenchymal Stem Cells Improves the Oxidative Stress Response and Mitochondrial Function in Rat Donation After Circulatory Death Livers

Abstract

There is a need to improve the quality of donor liver from donation after circulatory death (DCD). The purpose of this study was to investigate the effects and mechanism of normothermic machine perfusion (NMP) combined with bone marrow mesenchymal stem cells (BMMSCs) on the oxidative stress and mitochondrial function in DCD livers. DCD livers were obtained, a rat NMP system was established, and BMMSCs were extracted and identified. The DCD livers were grouped by their preservation method: Normal, static cold storage (SCS), NMP (P), and NMP combined with BMMSCs (PB), and the preservation time was up to 8 h. An IAR20 cell oxidative stress injury model was established in vitro by simulating DCD oxidative stress injury and coculturing with BMMSCs for 6 h. Compared with SCS group, after 6 h in vitro, the PB and P groups had significantly improved liver function and liver histological damage, reduced hepatocyte apoptosis and oxidative stress, improved hepatocyte mitochondrial damage, and increased mitochondrial membrane potential. These indicators were significantly better in the PB group than in the P group. BMMSCs significantly inhibited reactive oxygen species release from the IAR20 cell oxidative stress model in vitro, ameliorated mitochondrial damage, and increased mitochondrial membrane potential level. BMMSCs also downregulated the JUN N-terminal kinase-nuclear factor kappa B ( JNK-NF-kB) signaling pathway significantly in the IAR20 cell oxidative stress model and promoted AMP-activated protein kinase (AMPK) activation. We verified that NMP combined with BMMSCs also played the same role in the PB group. NMP combined with BMMSCs could improve liver quality by relieving oxidative stress injury and improving mitochondrial function in rat DCD livers. The mechanism of protective role might involve inhibiting the JNK-NF-kB pathway to reduce oxidative stress and promote AMPK activation, thereby reducing mitochondrial damage and increase mitochondrial function.

Protective Effects of Bone Marrow Mesenchymal Stem Cells (BMMSCS) Combined with Normothermic Machine Perfusion on Liver Grafts Donated After Circulatory Death via Reducing the Ferroptosis of Hepatocytes

“…The NMP system we used in this study was a self-modified single-cycle system. Based on our previous studies, the donor liver function was best at 6 h perfusion by the NMP system [ 20 , 27 ].…”

Heme Oxygenase-1-Modified Bone Marrow Mesenchymal Stem Cells Combined with Normothermic Machine Perfusion Repairs Bile Duct Injury in a Rat Model of DCD Liver Transplantation via Activation of Peribiliary Glands through the Wnt Pathway

“…The isolation, culture, identification, and gene transfection of BMMSCs were carried out as described previously [ 9 , 10 ]. Bone marrow contents were extracted from the rat femur and tibia, and BMMSCs were obtained by the continuous adhesion method [ 13 ]. Adenovirus vectors were used to transfect BMMSCs with constructs expressing the green fluorescent protein (GFP) and/or HO-1.…”

Combination of mesenchymal stromal cells and machine perfusion is a novel strategy for organ preservation in solid organ transplantation

“…The combination of MSCs with NMP resulted in the reduction of the levels of MPO and MDA and mitochondrial damage; increased the production of GSH; and ameliorated DCD liver oxidative stress injury. The study data also provided evidence that the normal functioning of the JNK/NF-κB and AMPK pathways was restored after MSC perfusion, both of which are known to be closely related to oxidative stress (Yang et al 2020a , 2020b ). Additionally, regardless of oxidative stress, disorders in the donated liver microcirculatory structure and function are other factors that considerably affect the liver physiological function post-transplantation (Vollmar and Menger 2009 ).…”

“…The same group reported that MSCs plus NMP pre-transplantation could also demonstrate this aspect of benefit. By inhibiting macrophage activation, by reducing microvascular leukocyte aggregation, by alleviating endothelial cell injury and by regulating ET-1/NO imbalance, this strategy effectively alleviates hepatic sinusoid congestion, maintains hepatic microcirculation homeostasis and favors the improvement of DCD liver quality (Yang et al 2020a , 2020b ).…”