Rat liver ischemia/reperfusion induced proinflammatory mediator and antioxidant expressions analyzed by gene chips and real-time polymerase chain reactions.
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.


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 alpha (TNFalpha). Expressions of TNFalpha, interleukin 12 (IL12), cyclooxygenase II (COXII), and other inflammatory mediators were analyzed by gene chips. COXII, TNFalpha, 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, TNFalpha, ALT, and LDH (P < .01) in the I/R but not the sham-operated group. Reperfusion induced significant increases in the expressions of TNFalpha, 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.
Hsu
Y C
YC
Department of Gastroenterology, Changhua Christian Hospital, Changhua, Taiwan.
Chou
T Y
TY
Chen
C F
CF
Wang
D
D
Su
C L
CL
Hu
R T
RT
D000818:Animals; D007511:Ischemia / Q000503:physiopathology*; D007770:L-Lactate Dehydrogenase / Q000235:genetics; D008102:Liver Circulation; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification; D051381:Rats; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000503:physiopathology*; D013482:Superoxide Dismutase / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics
D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D007770:L-Lactate Dehydrogenase; D013482:Superoxide Dismutase