A Multidrug Donor Preconditioning Improves Steatotic Rat Liver Allograft Function and Recipient Survival After Transplantation

Abstract

The scarcity of donors has prompted the growing utilization of steatotic livers, which are susceptible to injuries following orthotopic liver transplantation (OLT). This study aims to assess the efficacy of multidrug donor preconditioning (MDDP) in alleviating injuries of steatotic grafts following rat OLT. Lean rats were subjected to a Western-style diet with high-fat (HF) and high-fructose (HFr) for 30 days to induce steatosis. Both lean and steatotic livers were implanted into lean recipients fed with a chow diet after OLT. The HF + HFr diet effectively elevated blood triglyceride and cholesterol levels and induced fat accumulation in rat livers. Our results demonstrated a significant decrease in alanine aminotransferase levels (p = 0.003), aspartate aminotransferase levels (p = 0.021), and hepatic Suzuki scores (p = 0.045) in the steatotic rat liver allograft group following MDDP treatment on post-operation day (POD) 7. Furthermore, the survival rates of steatotic rat liver allografts with MDDP (19/21, 90.5%) were significantly higher than those in the steatotic control (12/21, 57.1%, *p = 0.019). These findings indicate that MDDP treatment improves steatotic rat liver allograft function and recipient survival following OLT.

Keywords: allograft function; donor shortage; ischemia reperfusion injury; multidrug donor preconditioning; orthotopic liver transplantation; rat steatotic liver donor.

FIGURE 1

Overall experimental design of the present study. (A) The flow chart of rat fatty liver model fed with chow or high fructose + high fat (HF + HFr) diets, multidrug donor preconditioning (MDDP) treatment, and rat orthotopic liver transplantation (OLT). (B) The rat blood and liver samples collection plan.

FIGURE 2

Macroscopic and microscopic features of rat liver allografts fed with chow and HF + HFr diets. The gross appearance of lean (A–C) and steatotic (D–F) rat liver allografts prior, post-HTK perfusion, and in cold storage. The body weight [BW, (G)], liver weight [LW, (H)], and LW/BW ratio (I) of lean and steatotic groups. The H&E staining images (×20) of lean (J) and fatty (K) rat liver allografts and quantitative analyses of steatosis [(L), n = 4 per group; *p < 0.05 or **p < 0.01, t-Test].

FIGURE 3

The clinical chemistry parameters of rat liver donor blood samples. The liver function panel includes ALT (A), AST (B), LDH (C), ALP (D), Bilirubin (E), Triglyceride (F), LDL (G), and HDL (H) of lean and steatotic rats fed with chow or HF + HFr diets shortly before liver explantation (n = 4 per group, *p < 0.05 or ***p < 0.001, t-Test).

FIGURE 4

The reperfusion of rat liver allografts and in vivo microcirculation study. The lean (A, B) and steatotic (C, D) allografts at 3 min and 15 min after reperfusion. (E, F) In vivo microcirculation studies of the rat liver allograft 24 h after transplantation showed the hepatic sinus, central vein (CV, 20x), and Hepatic leukocyte rolling (LR, 20x). (G–I) The quantitative analyses of sinusoidal reperfusion (G), sinusoidal leukostasis (H), and venule leukocyte adhesion [(I), n = 4 per group, *p < 0.05, t-Test].

FIGURE 5

The histological studies of rat liver allografts after transplantation. Histology analyses on POD 1 and 7 after transplantation, including Hepatocyte vacuolization (A), Hepatic architectures (B), Hepatic apoptosis (C), Hepatic necrosis (D), Hepatic fat contents (E), and Suzuki scores [(F), n = 3 or 4 per group, *p < 0.05, **p < 0.01, or ***p < 0.001, respectively, t-Test].

FIGURE 6

The clinical chemistry parameters of rat recipient blood samples. The liver function panel includes ALT (A), AST (B), LDH (C), ALP (D), Bilirubin (E), Triglyceride (F), LDL (G), and HDL (H) of lean and steatotic rats fed with chow or HF + HFr diets on POD 1 and 7 after OLT without or with MDDP (n = 3 or 4 per group, *p < 0.05, **p < 0.01 or ***p < 0.001, t-Test).

FIGURE 7

The rat recipient’s survival time after orthotopic liver transplantation (OLT). Rat recipient survival after OLT of lean (A) and steatotic (B) liver allografts with 8 h of cold ischemia and without or with MDDP treatment (n = 21 per group, *p < 0.05, Log-rank test).