AIM： To evaluate immunological protection of nitric ox- ide （NO） in hepatopulmonary syndrome and probable mechanisms of ischemia-reperfusion （IR） injury in rat liver transplantation, METHODS： Sixty-six healthy m...AIM： To evaluate immunological protection of nitric ox- ide （NO） in hepatopulmonary syndrome and probable mechanisms of ischemia-reperfusion （IR） injury in rat liver transplantation, METHODS： Sixty-six healthy male Wistar rats were randomly divided into three groups （11 donor/recipi-ent pairs）. In group 11, organ preservation solution was lactated Ringer＇s solution with heparin 10 000/μL at 4℃. In groups I and 111, the pLeservation solution added, respectively, L-arginine or Ng-L-arginine methyl ester （L-NAME） （1 mmol/L） based on group 11, and recipients were injected with L-arginine or L-NAME （50 mg/kg） in the anhepatic phase. Grafted livers in each group were stored for 6 h and implanted into recipi- ents. Five rats were used for observation of postopera- tive survival in each group. The other six rats in each group were used to obtain tissue samples, and execut- ed at 3 h and 24 h after transplantation. The levels of alanine aminotransferase （ALT）, tumor necrosis factor （TNF）-a and NO metabolites （NOx） were detected, and expression of NO synthase, TNF-α and intercellular adhesion molecule 1 （ICAM-1） was examined by tri- phosphopyridine nucleotide diaphorase histochemical and immunohistochemical staining. RESULTS： By supplementing L-arginine to strengthen the NO pathway, a high survival rate was achieved and hepatic function was improved. One-week sur- viral rate of grafted liver recipients in group I was significantly increased （28.8 4±36.6 d ys 4 4±1.7 d, P 〈 0.01） as compared with groups 11 and Ill. Serum levels of ALT in group ] were 2-7 times less than those in groups 11 and 11I （P 〈 0.01）. The cyclic guanosine monophosphate （cGMP） levels in liver tissue and NOx in group I were 3-4 times higher than those of group 11 after 3 h and 24 h reperfusion, while in group ]]], they were significantly reduced as compared with those in group ]1 （P 〈 0.01）. The levels of TNF-（z in group I were significantly lower than in group Ⅱ after 3 h and 24 h reperfusion （P 〈 0.01）, while being sig- nificantly higher in group Ⅲ than group Ⅱ （P 〈 0.01）. Histopathology revealed more severe tissue damage in graft liver and lung tissues, and a more severe in- flammatory response of the recipient after using NO synthase inhibitor, while the pathological damage to grafted liver and the recipient＇s lung tissues was signifi-cantly reduced in group I after 3 h and 24 h reperfu- sion. A small amount of constitutive NO synthase （cNOS） was expressed in liver endothelial cells after 6 h cold storage, but there was no expression of inducible NO synthase （iNOS）. Expression of cNOS was particularly significant in vascular endothelial cells and liver cells at 3 h and 24 h after reperfusion in group Ⅱ but expres- sion of iNOS and ICAM-1 was low in group I. There was diffuse strong expression of ICAM-1 and TNF-α in group Ⅱ at 3 h after reperfusion. CONCLUSION： The NO/cGMP pathway may be critical in successful organ transplantation, especially in treat- ing hepatopulmonary syndrome during cold IR injury in rat orthotopic liver transplantation.展开更多
文摘AIM： To evaluate immunological protection of nitric ox- ide （NO） in hepatopulmonary syndrome and probable mechanisms of ischemia-reperfusion （IR） injury in rat liver transplantation, METHODS： Sixty-six healthy male Wistar rats were randomly divided into three groups （11 donor/recipi-ent pairs）. In group 11, organ preservation solution was lactated Ringer＇s solution with heparin 10 000/μL at 4℃. In groups I and 111, the pLeservation solution added, respectively, L-arginine or Ng-L-arginine methyl ester （L-NAME） （1 mmol/L） based on group 11, and recipients were injected with L-arginine or L-NAME （50 mg/kg） in the anhepatic phase. Grafted livers in each group were stored for 6 h and implanted into recipi- ents. Five rats were used for observation of postopera- tive survival in each group. The other six rats in each group were used to obtain tissue samples, and execut- ed at 3 h and 24 h after transplantation. The levels of alanine aminotransferase （ALT）, tumor necrosis factor （TNF）-a and NO metabolites （NOx） were detected, and expression of NO synthase, TNF-α and intercellular adhesion molecule 1 （ICAM-1） was examined by tri- phosphopyridine nucleotide diaphorase histochemical and immunohistochemical staining. RESULTS： By supplementing L-arginine to strengthen the NO pathway, a high survival rate was achieved and hepatic function was improved. One-week sur- viral rate of grafted liver recipients in group I was significantly increased （28.8 4±36.6 d ys 4 4±1.7 d, P 〈 0.01） as compared with groups 11 and Ill. Serum levels of ALT in group ] were 2-7 times less than those in groups 11 and 11I （P 〈 0.01）. The cyclic guanosine monophosphate （cGMP） levels in liver tissue and NOx in group I were 3-4 times higher than those of group 11 after 3 h and 24 h reperfusion, while in group ]]], they were significantly reduced as compared with those in group ]1 （P 〈 0.01）. The levels of TNF-（z in group I were significantly lower than in group Ⅱ after 3 h and 24 h reperfusion （P 〈 0.01）, while being sig- nificantly higher in group Ⅲ than group Ⅱ （P 〈 0.01）. Histopathology revealed more severe tissue damage in graft liver and lung tissues, and a more severe in- flammatory response of the recipient after using NO synthase inhibitor, while the pathological damage to grafted liver and the recipient＇s lung tissues was signifi-cantly reduced in group I after 3 h and 24 h reperfu- sion. A small amount of constitutive NO synthase （cNOS） was expressed in liver endothelial cells after 6 h cold storage, but there was no expression of inducible NO synthase （iNOS）. Expression of cNOS was particularly significant in vascular endothelial cells and liver cells at 3 h and 24 h after reperfusion in group Ⅱ but expres- sion of iNOS and ICAM-1 was low in group I. There was diffuse strong expression of ICAM-1 and TNF-α in group Ⅱ at 3 h after reperfusion. CONCLUSION： The NO/cGMP pathway may be critical in successful organ transplantation, especially in treat- ing hepatopulmonary syndrome during cold IR injury in rat orthotopic liver transplantation.
