In vivo suppressive effect of nuclear factor-kappaB inhibitor on neutrophilic inflammation of grafts after orthotopic liver transplantation in rats.
OBJECTIVE
To investigate the effect of pyrrolidine dithiocarbamate (PDTC), a novel nuclear factor-kappaB (NF-kappaB) inhibitor, on expression of multiple inflammatory mediators and neutrophilic inflammation of cold preserved grafts after rat liver transplantation and its significance.


METHODS
Orthotopic liver transplantation (OLT) was performed after 24 h of cold storage using University of Wisconsin solution with varied concentrations of PDTC. We determined the time course of NF-kappaB activation and expression of multiple inflammatory signals, such as tumor necrosis factor-alpha (TNF-alpha), cytokine-inducible neutrophil chemoattractant (CINC), and intercellular adhesion molecule-1 (ICAM-1) by ELISA methods. Serum alanine aminotransferase (ALT), intrahepatic myeloperoxidase (MPO)/WBC (a measure of neutrophil accumulation) and Mac-1 expression (a measure of circulating neutrophil activity) were also evaluated.


RESULTS
PDTC decreased NF-kappaB activation induced by prolonged cold preservation in a dose dependent manner (from 20 mmol/L to 60 mmol/L), diminished TNF-alpha, CINC, ICAM-1 proteins in the grafts, and reduced the expression of increases in plasma TNF-alpha levels induced by prolonged cold preservation. Neutrophilic inflammation of the graft was significantly suppressed after preservation with PDTC (P<0.05). The total neutrophil accumulation in PDTC (40 mmol/L) group (7.04+/-0.97) was markedly reduced compared to control group (14.07+/-1.31) (P<0.05). Mac-1 expression was significantly reduced in PDTC (40 mmol/L) group (181+/-11.3%) compared with the control group (281+/-13.2%) (P<0.05) at 6 h after reperfusion. Furthermore, PDTC inhibited the increased serum ALT levels after liver transplantation.


CONCLUSIONS
PDTC can inhibit B NF-kappaB activation and expression of the inflammatory mediators, which are associated with improved graft viability via inhibiting intrahepatic neutrophilic inflammation. Our study suggests that a therapeutic strategy directed at inhibition of NF-kappaB activation in the transplanted liver might be effective in reducing intrahepatic neutrophilic inflammation, and would be beneficial to cold preserved grafts.
Gu
Xiao-Ping
XP
Department of Hepatobiliary Surgery, Drum Tower Hospital, Medical Department of Nanjing University, Nanjing 210008, Jiangsu Province, China.
Qiu
Yu-Dong
YD
Xu
Fu-Tao
FT
Jiang
Yong
Y
Ding
Yi-Tao
YT
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D019743:Chemokines, CXC / Q000097:blood; D006085:Graft Survival / Q000187:drug effects* / Q000276:immunology; D006505:Hepatitis / Q000188:drug therapy* / Q000276:immunology; D007166:Immunosuppressive Agents / Q000494:pharmacology; D018836:Inflammation Mediators / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000097:blood; D008099:Liver / Q000187:drug effects / Q000276:immunology; D016031:Liver Transplantation; D008297:Male; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000378:metabolism; D009504:Neutrophils / Q000276:immunology; D011759:Pyrrolidines / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D013859:Thiocarbamates / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D000975:Antioxidants; D019743:Chemokines, CXC; D007166:Immunosuppressive Agents; D018836:Inflammation Mediators; D036341:Intercellular Signaling Peptides and Proteins; D016328:NF-kappa B; D011759:Pyrrolidines; D013859:Thiocarbamates; D014409:Tumor Necrosis Factor-alpha; C020972:pyrrolidine dithiocarbamic acid