Therefore, we have hypothesized SAHA HDAC that the combination of these two drugs in donor animals could have a synergetic effect to decrease necrosis and apoptosis. Male Wistar rats weighing 280–350 g were used for both organ donors and recipients of the kidney graft. Animals were submitted to a 12-h day/night

cycle with access to water and standard laboratory chow ad libitum. All animal experiments were performed according to guidelines set by the US National Institutes of Health (NIH publication no. 28, revised 1996). We used tacrolimus (Prograf, Gador, Bs As, Argentina), medical grade, donated by Gador Argentina, and rapamycin (Sirolimus, Wyeth, Bs As, Argentina), medical grade, donated by Wyeth Argentina. Donor rats were anaesthetized KU-57788 chemical structure with intraperitoneal (i.p.) atropine 0·01 mg/kg, buprenorphine 0·04 mg/kg, diazepam 10 mg/kg

and, 10 min later, with ketamine 100 mg/kg body weight. The donors’ blood vessels and ureter were fully separated. Subsequently, the kidney was flushed via the aorta with 3 ml of 4°C cold Ringer lactate solution until it turned homogeneously pale. The left kidney was then removed with its vascular and ureteral pedicle and stored for 180 ± 15 min in cold Ringer lactate solution at 4°C. Recipients animals were nephrectomized bilaterally and underwent transplantation as described elsewhere [31,32]. Briefly, after flushing grafts with 5 ml normal Ringer’s solution, arterial and venous anastomoses were performed as end-to-side anastomoses to the aorta and inferior vena cava, respectively. Finally, the anastomosis of the ureter with the urinary bladder was constructed. The rat’s body temperature

was monitored and kept constant between 35 and 37°C in all cases. Rats were allowed to recover on a warm blanket with free access to water and standard laboratory chow ad libitum. Twenty-four hours after the transplant procedure, blood samples were C59 datasheet obtained for analysis, then animals were sacrified and kidneys were removed for histological evaluation. One dose of immunosuppressive drugs was administered to donor animals 12 h before nephrectomy. Doses and administration route were chosen according to previous reports [17]. Donor rats were divided randomly into four groups: Group 1 (control, n = 6): no immunosuppression was administered. Group 2 (rapa, n = 6): rapamycin (2 mg/kg, Sirolimus, Wyeth, Argentina) by gavage. Group 3 (FK506, n = 6): tacrolimus (0, 3 mg/kg, Prograf, Gador, Argentina) by gavage. Group 4 (rapa+ FK506, n = 6): tacrolimus (0, 3 mg/kg) + rapamycin (2 mg/kg) by gavage. None of the recipient animals received any immunosuppressive drug after transplantation. In addition, six rats underwent a sham procedure. Twenty-four hours before and after transplant the following blood determinations were performed: blood urea nitrogen (BUN), creatinine and C3 complement fraction (C3). C3 was measured by radial immunodiffusion and BUN and creatinine by ultraviolet kinetic and colorimetric-kinetic, respectively (Mindray 300).