Corticosteroids Enhance Hepatic Injury Following Ischemia- Reperfusion P. Sileri, S. Schena, J. Fukada, C. Rastellini, J. Pirenne, E. Benedetti, and L. Cicalese H ISTORICALLY, in solid organ transplantation, cor- ticosteroids (CCS) have been used widely as an integral part of the immunosuppression protocols. How- ever, due to long-term CCS side effects, as well as the availability of newer and more potent immunosuppressive agents, the role of CCS has been reassessed. Recently, CCS-free immunosuppressive protocols have been success- fully proposed for liver and kidney transplantation. 1,2 Al- though these approaches seem safe when compared with historic controls, the majority of transplant programs are still wary of their introduction. 3,4 Nevertheless, the role of CCS in ischemia-reperfusion injury following transplanta- tion has not been adequately investigated. Because data on the effect of CCS on I-R injury are scant, the present study aims to investigate whether CCS admin- istration interferes with I-R–induced hepatic injury in rats. MATERIALS AND METHODS Male ACI rats (Sprague-Dawley, Inc, Indianapolis, Ind), weighing 200 to 250 g, were used in this study. Animals were randomly divided into two groups. One hour prior to partial liver ischemia study animals were treated with 2 mg/kg intravenously (IV) of methylprednisolone, whereas control animals received the same volume of IV saline solution. In each group (n = 4), partial liver ischemia was done by clamping the portal and arterial branches to the left lateral and median lobes of the liver for 60 minutes. Animals were killed at 2, 6, 24, and 48 hours after reperfusion. Serum AST, ALT, and LDH were measured and liver samples were obtained and examined for morphology, DNA fragmentation (DNA laddering), and evaluation of apoptosis (TUNEL). Statisti- cal analyses were performed using Student’s t test and the Mann– Whitney U test. RESULTS All the animals survived the procedure. Enzymatic levels were increased significantly, confirming liver damage in both groups. In the CCS-treated animals AST levels were significantly higher at 24 hours compared with controls (1354  183 vs 746  103; P  .02). Histologic alteration of the liver samples obtained after I-R from the control animals was characterized as diffuse cytoplasmic vacuoliza- tion/degeneration and congestion during the early phases of reperfusion (2 and 6 hours). Sinusoidal dilation and con- gestion were enhanced later (24 hours). In treated animals the injury was qualitatively similar but slightly increased. Different patterns of DNA fragmentation were observed. In the control group, DNA ladders were concentrated mostly after 2 hours of reperfusion and were still moderately evident after 24 hours, but disappeared by 48 hours. In CCS-treated rats, DNA fragmentation was increased com- pared with controls at 2 and 6 hours. Apoptosis was significantly increased following 2 and 6 hours of reperfu- sion in the CCS-treated group (2 hours: 56  2.7 vs 72  2.4; 6 hours: 57  2 vs 63  1.9; data expressed as mean TUNEL-positive nuclei/field observed  SEM, control vs CCS-treated, respectively; P  .05). CONCLUSIONS Data obtained from this study suggest that a single dose of CCS enhances hepatic I-R injury. This effect is mediated by increased DNA fragmentation, and apoptosis is observed soon after reperfusion. These data suggest that negative side effects of CCS are limited not only to the long term but probably also take place immediately, during reperfusion. REFERENCES 1. Punch JD, Shieck VL, Campbell DA, et al: Surgery 118:783, 1995 2. Tisone G, Angelico M, Palmieri G, et al: Transplantation 67:1308, 1999 3. Birkeland SA: Transplantation 71:1089, 2001 4. Reding R: Transplantation 70:405, 2000 From the Division of Transplant Surgery, University of Illinois at Chicago, Chicago, Illinois, USA. Address reprint requests to Dr Pierpaolo Sileri, Division of Transplant Surgery, 840 South Wood Street, Room 402, Chi- cago, IL 60612. E-mail: ppsileri@uic.edu. 0041-1345/01/$–see front matter © 2001 by Elsevier Science Inc. PII S0041-1345(01)02514-3 655 Avenue of the Americas, New York, NY 10010 3712 Transplantation Proceedings, 33, 3712 (2001)