Cho, WH

Kim, DG

Mischinger, HJ

Murase, N

Starzl, TE

Todo, S

English

PubMed

D-Scholarship@Pitt

KA~t 1990 BRIEF COMMt:SICATIONS 353 i~ enzyme release and decrease in bile flo .......... ith time (3). The change in ..... eight and change in platelets were compared using Student's t test as were the means for perfusate flow and oxygen consumption (Table 1). The latter parameters do not show exponential changes in control unpreserved livers (3). After analysis, it was found that none of the parameters consistently differed between the groups. Particularly, there was no deterioration in bile flow or increase in enzyme release into the perfusate after flushing with Ringer's lactate (Table 1). Both these parameters are good markers of hepatic allograft viability in this model (3). Sundberg et aI., using an isolated rabbit liver, have shown that bile production was significantly better, following 24-hr storage in UW solution, in livers flushed with UW solution as opposed to those flushed with other solutions including Ringer's lactate. They were unable to dem-onstrate any difference in AST concentrations between the groups (6). In our study perfusate flow was significantly better at one time point with UW flushing, and there was a trend toward better perfusate flow at two other time points. We have previously shown that the key injury of cold preservation is to the microcirculation (7), and it is possible that the observations made in respect to perfusate flow might be important in a large animal model, despite the failure to find consistent differences in enzyme release and bile flow in this model. Because UW solution is a superior preservation solution, one cannot a priori conclude that it should be used for organ flushing, but there is enough suggestive evidence that it might be better to justify a simple trial in clinical transplantation. G. R. MORGAN2 P. R. C. HARVEyJ S. M. STRASBERG' Department of Surgery Mt. Sinai Hospital Research Institute University of Toronto Lil.'f!r Transplantation Progr U · . f~ am nlverslty 0 loronto Toronto, Canada M5G 1 X5 2 Dr. Morgan is a fellow of the Canadian Liver Foundation. 3 Dr. Harvey is an Ontario Ministry of Health Scholar. , Address correspondence to: Dr. S. M. Strasberg 1225·600 Univer. sity Avenue, Toronto, Ontario, Canada M5G lX5. REFERENCES 1. Belzer FO. Clinical organ preservation with UW solution. Trans· plantation 1989; 47: 1097. 2. Collin GM. New approaches for preservation. Fourth International Symposium on Organ Procurement and Preservation. September 17-20, 1989. Minneapolis, Minnesota. 3. lu S, Harvey PRC, Mako",'ka L. Petrunka CN. llson RG, Strasberg SM. Markers of allograft viability in the rat. Transplantation 1987; 44: 562. 4. Todo S. Podesta L, Ueda Y. et al. Comparison of UW with other solutions for liver preservation in dogs. Clin Transplant 1989; 3: 253. 5. Steffen R. Ferguson D. Ludwig J, Kromm RAF. Efficacy of Euro· Collins and University of Wisconsin solutions in rat liver pres· ervation assessed by orthotopic liver transplantation. Eur Surg Res 1989; 2l(suppl 2): 9. 6. Sundberg R. Ar'Rajab A. Ahren B, Bengmark S. Effect of initial cooling solution on 24 hour liver preservation with UW solution. Clin Transplant 1989; 3: 290. 7. McKeown CMB. Edwards V, Phillips MJ, Harvey PRC, Petrunka CN, Strasberg SM. Sinusoidal lining cell damage: the critical injury in cold preservation of liver allografts in the rat. Trans· plantation 1988; 46: 178. Received 28 November 1989. Accepted 22 February 1990. COMPARISON OF SUPEROXIDE DlSML'TASE, ALLOPURINOL, COENZYME QI0, AND GLUTATHIONE FOR THE mobsb~qflk OF WARM ISCHEMIC INJURY' There have been many reports that the administration of free radical scavenger compounds can interdict oxygen free radicals and limit this cause of cell damage. Four such drugs, superoxide dismutase (1, 2), allopurinol (3), coenzyme QI0 (4), and glutathione (5) ha\'e been evaluated in different organs using a variety of experimental models, but there is little information about their comparative value. In this study, we have attempted to provide such information in male Lewis rats weighing 220-260 g. The animals were given water ad libitum for 12 h, anesthetized by intraperitoneal injection of 40 mglkg sodium pentobarbital (Nembutal), and submitted to cross-damping of the hepatic arterial and portal venous branches to the lateral and median lobes of the liver_ This procedure (6) imposes a standard ischemic insult on about two-thirds of the liver, without causing splanchnic venous stasis since portal blood flow is well accommodated by the right and right caudate liver lobes that are left perfused. The perfused hepatic fraction is large enough to prevent hepatic failure and consequent deterioration of the animal during the ensuing 24 hr, after which the abdominal incision was reopened_ Blood was taken 1 Tbia work was 8Upported by Research Grants (rom the Veterans Adminiatration and by Project Grant OK 29961 from the National IDatitutea of Health, Betheeda, MD. from the abdominal aorta for liver function tests, and the animals were killed. The extent of ischemic necrosis in the damaged liver fragment was measured by making 3 thin (less than 2·mm) slices from each devascularized lobe and by stain· ing these with tetranitro blue tetrazolium (TNBT. Sigma Cor-poration, Sigma Chemical Company, St. Louis, MO) in a phos-phate buffer solution at pH 7.4 (6). The stained necrotic margin was measured by tracing it with a computer digitizer and Summa Sketch MM1201. In c~ntrol experiments, it voas shown that crossclamping for 150 mm was necessary before a high degree of necrosis was produced in the devascularized lobes. This was determined with biochemical and morphologic criteria (Fig. 1) in 36 experiments using crossclamp times of 30, 50, 90, 120, 150, and 180 min (n=6 at each time). SGOT and necrosis data from the ISO-min crossciamping were the controls used for comparison with the test groups (Table 1). Drugs tested were: (1) superoxide dismutase (human Du/Z SOD, Pharmacia AB Uppsala, Sweden), (2) allopurinol (Aldrich Chemical Company. Inc., Milwaukee, WI), (3) coenzyme QIO (Eisai Company, Tokyo, Japan). and (4) a reduced form of glutat~ione (Aldrich Chemical Company, Inc., Milwaukee, WI). For each of these agents, 3 doses were tried (Table 1). The drugs were administered intravenously into the internal jugular 3M TRANSPLANT ATJON Vol so. No. ~ ,.. 80 # : 60 .. ~ . ~ 40 o .. u ~ 20 , _ ...-:;-::-::i - - • -/,f , , , , , . , , , , , 14 12 10 ..J __ .... 0 8 ;:)0 6 1- 0 0-4 C) )( rn ...... 2 • , , I --- _. --o ~~~ ____ ~ __ ~ ____ ~ __ ~-KKg 0 • 30 60 90 120 150 180 Warm Ischemia Time (min) FlGl"ltE 1. Relation of cro88Clamp time to the area of necroaia in the liver slices and to SGOT rise. Mean ±SE for the ISO· min iachemia data are eiven in Table 1. (n=6 at each time.) TABLE 1. Dose·effective test of various scavengers with 150 min ischemia Douce SGOT(U!L) Necrotic pcaven~r n &re4 (%) (mg/kc) (mean %SE) (mean %8E) No treatment 0 6 7193.1±968.2 62.6±7.0 SOD 10 6 4676.0±938.9 24.9±3.5 20" 6 2724.3±724,4 12.2±1.4 30 5 2737.8±80U 20.9±4.2 Allopurinol 50 5 873I.6±2107.5 64.7±8.l 100 6 5182.'±1078.1 38.1±7.8 200" • 6302.7± 1908. 1 38.0±6.3 Coenzyme QIO 5 6 11,549.3±1852.' 73.7±7.0 10" 6 7806.7±1696.9 56.2±4.8 20 5 12,600.0±1506.7 52.6±8.8 Glutathione 250 • 13,273.3±2811.6 72.5±6.1 500" 6 994 1.3± 1404.5 75.0±3.8 1000 5 14,518.4±1862.7 96.7±O.7 ° Most effective dose used for experimenU shown in Figure 2. vein 5 min (SOD) or 60 min (allopurinol, co·enzyme, and gluathione) before beginning 150 min of ischemia. The most effective of these doses (Table 1) were used for the more complete nperiments summarized in Figure 2. Data were ex-pressed as mean ±SE. Comparison of group means was with the Student's t test (unpaired), in which a probability of error less than 0.025 was considered as significant. Superoxide dis mutase was the most effective agent. The rises in SGOT were reduced with SOD treatment before ischemia, or before reperfusion; treatment at both times was less effective than at either time alone (Fig. 2). The extent of necrotic tissue was significantly reduced with all 3 variations of timing, but the effect was most pronounced with the pre ischemia dose alo~ (Fig. 2). Allopurinol reduced the SGOT rise only if given before ischemia (Fig. 2). However, the area of necrosis was reduced with all 3 variations of treatment timing. Coenzyme QIO may have reduced the SGOT rise when given before reperfusion but there was no significant effect on the extent of necrosis with any of the treatment variations (Fig. 2). With gluathione treatment, the SGOT rises were higher than in untreated animals but the extent of measured iachemic necrosis was not influenced (Fig. 2). The superoxide anion, hydrogen peroxide, and the hydroxyl radical are the most important free radicals produced at the time of reperfusion. At this time, molecular oxygen is used to convert ischemic accumulations of hypoxanthine to xanthine (7). Because the oxygen free radicals may act on the endothelial 12 .... g 10 o .. a IC ..., 8 2 o 100 ; 80 -• ! 80 c ~ 40 o .. u : 20 o SOD Allop CoO Glutatll • a.,o,. Ilell.",I. o a.,o,. lIotll SCAVENGER m a.,o,. ,epe,'ullo. m Control IpcO.028 FlCURE 2. Effect of !!Cavengers on SGOT levels and on necrotic area, 24 hr after lSO·min of i.8cbemia. Agents were administered intra· venously 5 min (SOD) or 60 min (allop., CoQ. glutath.l hefoM! ischemia, 5 min herOM! reperfu.sion (all' &genU). or both. (SOD) superolide dismutase; (Allop) allopurinol; (CoQ) coenzyme QIO; (glutath) gluta. thione; (UIL) units per liter; (O)P<O.025 compared with controls.) cells, causing their swelling and increased permeability (7, 8), the scavenger drugs theoretically could protect the graft micro· vasculature. The most effective of the four scavenger drugs in our study was superoxide dis mutase, which to our surprise was most potent when given before the ischemic injury. Most pre· vious reports have suggested that SOD will not be effective unless it is given just before reperfusion (9). An effect of treatment at this time also was seen in our studies, but it was no stronger than treatment 5 min before the ischemia. SOD given before reperfusion did not add to the benefit of the preischemic treatment. A benefit with allopurinol was also noted. confirming the work of others (3, 10) but this was far less striking. Since coenzyme QI0 is known to have antioxidant as well as membrane-stabilizing properties, and has been said to protect from reperfusion injury (4), it was particularly dis· appointing to see no ameliorating effect of this agent on the extent of tissue necrosis. The same was true of glutathione, which is said to have a direct antioxidant effect (5). Caution is necessary in reaching conclusions in the rat either about the efficacy or lack thereof of any agent as it may apply to the human liver. Southards et al. (11) have pointed out that the rat may not be 8 suitable animal for research on free radical scavengers. They noted that the ratio of superoxide dismutase to xanthine oxidase in the rat liver was much lower than in the dog or human and suggested that the rat liver could be more sensitive to oxygen-derived free radical damage than in other species. H 80, positive results from SOD therapy could be too easily obtained in the rat and could overestimate what might ·be expected in humans. Species differences also could be re-sponsible for underrating other potentially useful substances, such as coenzyme QIO, glutathione, and other drugs classified as scavengers but with a biochemical mechanism different from SOD. WON-HvUN CHO DONG-GOO KIM NORIKO ML'RASE HANS-JORG MISCHINGER SATORU To DO THOMAS E. STARZL2 The Department of Surgery University Health Center of Pittsburgh University of Pittsburgh The Veterans Administration Medical Center Pittsburgh, PenrlS)b.'ania I This work was supported by Research Grants from the Veterans Administration and by Project Grant DK 29961 from the National Institutes of Health, Bethesda, MD, 2 Address correspondence to: Thomas E. Stanl, M.D., Ph.D., De-partment of Surgery, 3601 Fifth Avenue, Falk Clinic, Pittsburgh, PA 15213. REFERENCES 1. Atalla SL, Toledo-Pere)Ta LH, MacKenzie GH, et a1. Influence of oxygen -derived free radical scavengers on ischemic livers. Trans-plantation 1985; 40: 584. 35S 2. Koyarnadil, a1Bu~klevedKidBIK Wlt'lhliams ~~:K et .al. The role 01 ol),pn free ra C 810m at 109 e repe'lUlIlon Injury of cold ed ischemic kidney. Transplant.ation 19S5; 40: 590. preaerv 3. Castillo M. Toledo-Pereyra LH, Prough D, et al Eff- t' . . .... . .' ,.<IVe Imln, of allopunnol admmlstratlOn In the I!lChemlc liver Tra I tation 1989; 47: 727. . nap an· 4. Marubayashi S, Dohi K, Ezaki H, et al. Preservation of iachemic rat liver mitochondrial functions and liver viability with coen-zyme QI0. Surgery 1982; 91: 631. 5. McCay PB. Gibson DD, Fong KL. et al. Effett of glutathione peroxidase activity on lipid peroxidation in biological mem-branes. Biochem Biophys Acta 1976; 431: 459. 6. Frederiks WM. Fronik GM, Hesseling g~dK A method for quan-titative analysis of the extent of necrosis in ischemic rat liver. Exp Mol Pathol 1984; 41: 119. .. McCord JM:. Oxygen derived free radicals in postischemic tissue injury. ~ Engl J Med 1985; 312: 159. 8. Bulkley GB. The role of oxygen free radicals in human diseue processes. Surgery 1983; 94: 407. 9. Hoshino T. ~1aley WR, Bulkley GB, et al. Ablation of free radical-mediated reperfusion injury for the sal\'age of kidneys taken from non-heartbeating donors. Transplantation 1988; 45: 284. 10. Adkinson D. Hollwarth ME, Benoit IN, et al. Role of fr~ radicals in ischemia·reperfusion injury to the liver. Physiol Scand 1986; 548: 101. 11. Southard JH, Marsh DL, McAnulty JF, et al. The importance of O. derived free radical injury to organ preservation and trans-plantation. Transplant Proc 1987; 19: 1380. Received 17 August 1989. Accepted 29 January 1990. 

Comparison of superoxide dismutase, allopurinol, coenzyme Q10, and glutathione for the prevention of warm ischemic injury

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Comparison of superoxide dismutase, allopurinol, coenzyme Q10, and glutathione for the prevention of warm ischemic injury

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