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

Abstract

Livers from donors after circulatory death (DCD) are inevitably exposed to a longer warm ischemic period, which might increase the incidence of postoperative bile duct complications. Bone marrow mesenchymal stem cells (BMMSCs) have tissue repair properties. The present study was aimed at exploring the repair effect of heme oxygenase-1- (HO-1-) modified BMMSCs (HO-1/BMMSCs) combined with normothermic machine perfusion (NMP) on bile duct injury after DCD liver transplantation and at revealing the underlying mechanisms. Rat livers were exposed to in situ warm ischemia for 30 min; then, NMP was performed through the portal vein for 4 h with BMMSCs, HO-1/BMMSCs, or neither before implantation. Obvious bile duct histological damage and liver functional damage were observed postoperatively. In the group treated with HO-1/BMMSCs combined with NMP (HBP group), liver functions and bile duct histology were improved; meanwhile, cell apoptosis was reduced and cell proliferation was active. A large number of regenerative cells appeared at the injured site, and the defective bile duct epithelium was restored. Dilatation of peribiliary glands (PBGs), proliferation of PBG cells, high expression of vascular endothelial growth factor (VEGF), and increased proportion of bile duct progenitor cells with stem/progenitor cells biomarkers were observed. Blocking Wnt signaling significantly inhibited the repair effect of HO-1/BMMSCs on bile duct injury. In conclusion, HO-1/BMMSCs combined with NMP were relevant to the activation of biliary progenitor cells in PBGs which repaired bile duct injury in DCD liver transplantation via the Wnt signaling pathway. Proliferation and differentiation of PBG cells were involved in the renewal of the injured biliary epithelium.

Intestinal Microbiota Participates in the Protective Effect of HO-1/BMMSCs on Liver Transplantation With Steatotic Liver Grafts in Rats

“…MSCs play an important role in the repair and regeneration of tissue and organ damage and good results have been achieved in organ transplantation and repair (Yang et al, 2014;Yin et al, 2017Yin et al, , 2019Cao et al, 2020;Tian et al, 2021). Our research group has verified that HO-1-modified BMMSCs have anti-inflammatory, anti-oxidative, anti-apoptotic, and delayed rejection functions in transplant recipients, and are more effective than BMMSCs alone.…”

“…Our research group has verified that HO-1-modified BMMSCs have anti-inflammatory, anti-oxidative, anti-apoptotic, and delayed rejection functions in transplant recipients, and are more effective than BMMSCs alone. Activity in the diseased part is related to prolonging its duration of function (Cao et al, 2020;Tian et al, 2021). However, there is no relevant research on the effect of MSCs on steatotic liver transplantation.…”

miR-29a-3p in Exosomes from Heme Oxygenase-1 Modified Bone Marrow Mesenchymal Stem Cells Alleviates Steatotic Liver Ischemia-Reperfusion Injury in Rats by Suppressing Ferroptosis via Iron Responsive Element Binding Protein 2

“…Paraffin-embedded sections of liver tissue were subjected to immunohistochemical staining as described previously [ 27 ]. Primary antibodies recognizing 4-hydroxynonenal (4-HNE; 1 : 50, Abcam Inc., Cambridge, UK), CD68 (1 : 200, Santa Cruz Biotechnology, Dallas, TX, USA), myeloperoxidase (MPO) (1 : 200, ProteinTech), and IREB2 (1: 200, Santa Cruz Biotechnology) were incubated overnight at 4°C, followed by incubation with the secondary antibody and horseradish peroxidase-labeled streptavidin (HRP).…”

“…Total proteins were extracted from liver tissue samples, and the protein concentration was determined using the bicinchoninic acid (BCA) method. The standard western blotting method [ 27 ] was used with primary antibodies recognizing glutathione peroxidase 4 (GPX4; 1 : 500, ProteinTech), high mobility group box 1 (HMGB1; 1 : 500, ProteinTech), Toll-like receptor 4 (TLR4; 1 : 500, Santa Cruz Biotechnology), RELA proto-oncogene, NF- κ B Subunit (p65; 1 : 500, Cell Signaling Technology, Danvers, MA, USA), phosphorylated (p)-p65 (1 : 500, Cell Signaling Technology), IREB2 (1 : 200, Santa Cruz Biotechnology), FTH1 (1 : 500, Abcam), and TFR1 (1 : 200, Santa Cruz Biotechnology). Antibodies (1 : 800, Santa Cruz Biotechnology) detected β -actin as an internal reference protein.…”

Biliary Viability Assessment and Treatment Options of Biliary Injury During Normothermic Liver Perfusion—A Systematic Review

“…Additionally, oxidative stress markers, macrovesicular steatosis, and tissue triglycerides were reduced. Tian et al discovered that administration of Heme Oxygenase-1-modified bone marrow mesenchymal stem cells (HO-1/BMMSCs) during NMP of rat livers lead to improved liver function, bile duct histology, restored epithelium, and reduced cell apoptosis ( 50 ). Haque et al investigated the reconditioning of discarded DCD livers with tissue plasminogen activator administration during NMP.…”