Zinc-mediated reduction of apoptosis in cardiac allografts.
BACKGROUND
Apoptosis is thought to occur during immune-mediated acute rejection of cardiac allografts. In vitro studies have shown that zinc inhibits the activity of the proapoptotic enzyme caspase-3. We hypothesized that ZnCl(2) would reduce acute cardiac rejection in vivo via the blockade of caspase-3-dependent apoptosis. (99m)Tc-labeled annexin V was used to measure apoptosis in cardiac allografts through nuclear imaging. Annexin V binds to phosphatidylserines, which are externalized to the outer membrane of apoptotic cells.


RESULTS
Twenty-seven PVG rat hearts were transplanted heterotopically into the abdomen of untreated ACI rats as controls (group 1). Fifteen were scanned and euthanized on postoperative day 4, and 12 were assessed for graft survival. Group 2 and 3 rats (n=15 each) received 1 and 5 mg/kg ZnCl(2) BID IP, respectively. Nine of each of these groups were scanned and euthanized on postoperative day 4, and 6 were studied for allograft survival. Group 4 rats (n=3) received isografts. Region-of-interest analysis demonstrated a dose-dependent reduction in (99m)Tc annexin uptake in ZnCl(2)-treated allografts: 2.43+/-0.37% for group 1, 1. 97+/-0.41% for group 2, 1.21+/-0.47% for group 3, and 0.55+/-0.19% for group 4 (ANOVA, P:=0.001). Graft survival times of 6.4+/-1.7, 9. 3+/-3.0, and 11.5+/-3.4 days for groups 1, 2, and 3, respectively, were also observed (ANOVA, P:=0.001). Caspase-3 activity in the allografts showed a 3.7-fold reduction in group 3 animals compared with group 1 animals (P:=0.004).


CONCLUSIONS
Apoptosis that occurs in acute cardiac allograft rejection is reduced with ZnCl(2) in a dose-dependent manner via caspase-3 inhibition.
Kown
M H
MH
Department of Cardiothoracic Surgery, Stanford University School of Medicine, Stanford, CA 94305-5247, USA.
Van der Steenhoven
T
T
Blankenberg
F G
FG
Hoyt
G
G
Berry
G J
GJ
Tait
J F
JF
Strauss
H W
HW
Robbins
R C
RC
D000818:Animals; D017304:Annexin A5 / Q000032:analysis; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism; D002712:Chlorides / Q000097:blood / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D006085:Graft Survival / Q000187:drug effects; D006321:Heart / Q000000981:diagnostic imaging; D016027:Heart Transplantation; D007928:Lethal Dose 50; D008297:Male; D009206:Myocardium / Q000166:cytology / Q000201:enzymology; D015609:Organotechnetium Compounds / Q000032:analysis; D011877:Radionuclide Imaging; D019275:Radiopharmaceuticals / Q000032:analysis; D051381:Rats; D011912:Rats, Inbred ACI; D014184:Transplantation, Homologous; D017967:Zinc Compounds / Q000097:blood / Q000494:pharmacology*
D017304:Annexin A5; D061945:Caspase Inhibitors; D002712:Chlorides; D015609:Organotechnetium Compounds; D019275:Radiopharmaceuticals; D017967:Zinc Compounds; C417036:technetium Tc 99m annexin V; C016837:zinc chloride; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases