Delivery of the Bioactive Gas Hydrogen Sulfide During Cold Preservation of Rat Liver: Effects on Hepatic Function in an Ex vivo Model

Abstract

The insults sustained by transplanted livers (hepatectomy, hypothermic preservation, and normothermic reperfusion) could compromise hepatic function. Hydrogen sulfide (H₂S) is a physiologic gaseous signaling molecule, like nitric oxide (NO) and carbon monoxide (CO). We examined the effect of diallyl disulfide as a H₂S donor during hypothermic preservation and reperfusion on intrahepatic resistance (IVR), lactate dehydrogenase (LDH) release, bile production, oxygen consumption, bromosulfophthalein (BSP) depuration and histology in an isolated perfused rat liver model (IPRL), after 48 h of hypothermic storage (4 °C) in University of Wisconsin solution (UW, Viaspan). Livers were retrieved from male Wistar rats. Three experimental groups were analyzed: Control group (CON): IPRL was performed after surgery; UW: IPRL was performed in livers preserved (48 h-4 °C) in UW; and UWS: IPRL was performed in livers preserved (48 h-4 °C) in UW in the presence of 3.4 mM diallyl disulfide. Hypothermic preservation injuries were manifested at reperfusion by a slight increment in IHR and LDH release compared with the control group. Also, bile production for the control group (1.32 µL/min/g of liver) seemed to be diminished after preservation by 73% in UW and 69% in UW H₂S group at the end of normothermic reperfusion. Liver samples analyzed by hematoxylin/eosin clearly showed the deleterious effect of cold storage process, partially reversed (dilated sinusoids and vacuolization attenuation) by the addition of a H₂S delivery compound to the preservation solution. Hepatic clearance (HC) of BSP was affected by cold storage of livers, but there were no noticeable differences between livers preserved with or without diallyl disulfide. Meanwhile, livers preserved in the presence of H₂S donor showed an enhanced capacity for BSP uptake (k(A) CON = 0.29 min⁻¹; k(A) UW = 0.29 min⁻¹ ; k(A) UWS = 0.36 min ⁻¹). In summary, our animal model suggests that hepatic hypothermic preservation for transplantation affects liver function and hepatic depuration of BSP, and implies that the inclusion of an H₂S donor during hypothermic preservation could improve standard methods of preparing livers for transplant.

The Novel N,N‐bis‐2‐Hydroxyethyl‐2‐Aminoethanesulfonic Acid–Gluconate–Polyethylene Glycol–Hypothermic Machine Perfusion Solution Improves Static Cold Storage and Reduces Ischemia/Reperfusion Injury in Rat Liver Transplant

“…In group 2 (HTK), livers were stored for 24 hours in HTK solution at 0°C (n = 5), and in group 3 (BGP‐HMP), livers were stored for 24 hours in BGP‐HMP solution at 0°C (n = 5). After 24 hours of SCS, livers from groups 2 and 3 were reperfused in the IPRL at 37°C with KHB and a 2% dextran buffer …”

“…Portal venous pressure was measured during perfusion by means of a water column connected to the portal inflow line. Hepatic effluent was collected through a steel catheter placed firmly into the suprahepatic inferior vena cava and drained to the reservoir …”

The effect of a hydrogen sulfide releasing molecule (Na2S) on the cold storage of livers from cardiac dead donor rats. A study in an ex vivo model

“…In brief, 0.1 mL of saline containing 1000 UI of Heparin (Sobrius, Argentina) was injected into the femoral vein, the bile duct was cannulated with a PE-50 catheter (Intramedic, USA) and a 14 G Abbocath catheter was introduced and fixed in the portal vein. Perfusion with KHB was started immediately at 15 mL/min, the suprahepatic inferior vena cava was cannulated with steel tubing (internal diameter 3 mm) and, after blood wash out, the liver was carefully removed [2].…”

Medical Gases: A Novel Strategy for Attenuating Ischemia—Reperfusion Injury in Organ Transplantation?

“…This artificial means of inducing a hypothermic-like state had no apparent adverse effects; thus, the potential exists to take advantage of this reduced metabolic usage to improve organ preservation. When an H 2 S donor was added to the preservation solution in an ex vivo rat liver model, the damaging effects of cold storage were partially reversed [49]. In a mouse model of renal transplantation, treatment with H 2 S during the hypoxic stage of IRI resulted in dramatically reduced renal damage (Figure 5) [49, 50].…”

“…When an H 2 S donor was added to the preservation solution in an ex vivo rat liver model, the damaging effects of cold storage were partially reversed [49]. In a mouse model of renal transplantation, treatment with H 2 S during the hypoxic stage of IRI resulted in dramatically reduced renal damage (Figure 5) [49, 50]. The apparent mechanism for prevention of renal injury is by reversible inhibition of cytochrome c oxidase, the terminal enzyme of the mitochondrial electron transport chain [1, 50].…”