Rat Liver Ischemia/Reperfusion Induced Proinflammatory Mediator and Antioxidant Expressions Analyzed by Gene Chips and Real-Time Polymerase Chain Reactions

• Published in: Transplantation proceedings 2008-09, Vol.40 (7), p.2156-2158
• Main Authors: Hsu, Y.C, Chou, T.Y, Chen, C.F, Wang, D, Su, C.L, Hu, R.T
• Format: Article
• Language: English
• Subjects: Animals; Biological and medical sciences; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Gastroenterology. Liver. Pancreas. Abdomen; Ischemia - physiopathology; L-Lactate Dehydrogenase - genetics; Liver Circulation; Liver. Biliary tract. Portal circulation. Exocrine pancreas; Male; Medical sciences; Oligonucleotide Array Sequence Analysis; Other diseases. Semiology; Polymerase Chain Reaction; Rats; Rats, Sprague-Dawley; Reperfusion Injury - physiopathology; RNA, Messenger - genetics; RNA, Messenger - isolation & purification; Superoxide Dismutase - genetics; Surgery; Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases; Tissue, organ and graft immunology; Tumor Necrosis Factor-alpha - genetics
• ISSN: 0041-1345; 1873-2623
• DOI: 10.1016/j.transproceed.2008.07.088

Abstract Objective Ischemia/reperfusion (I/R) of the rat liver induces injury; however, few studies have investigated gene expressions associated with this phenomenon. In this study, gene chip and real-time polymerase chain reactions (PCR) were used to study the expressions of the proinflammatory mediators and antioxidants after I/R. Materials and Methods Ischemia was induced by clamping the common hepatic artery and portal vein for 40 minutes followed by 90 minutes reperfusion. Blood samples collected before ischemia and after reperfusion were analyzed for alanine amino transferase, lactic dehydrogenase, hydroxyl radicals, nitric oxide (NO), and tumor necrosis factor α (TNFα). Expressions of TNFα, interleukin 12 (IL12), cyclooxygenase II (COXII), and other inflammatory mediators were analyzed by gene chips. COXII, TNFα, and antioxidants of mitochondrial superoxide dismutase (SOD(Mn)), catalase, and heat shock protein 70 (HSP70) were double confirmed by real-time PCRs. Results This protocol resulted in elevations in the blood concentrations of NO, hydroxyl radicals, TNFα, ALT, and LDH ( P < .01) in the I/R but not the sham-operated group. Reperfusion induced significant increases in the expressions of TNFα, IL12, COXII, SOD(Mn), catalase, and HSP70. Real-time PCR also demonstrated increases in mRNA expressions of the proinflammatory mediators and antioxidants. Conclusions This protocol resulted in oxidative stress, nitrosative stress, and liver injury. The increases in expressions of both proinflammatory mediators and antioxidants suggested that an imbalance between inflammation and anti-inflammation could be the possible reason for the liver injury after I/R.