Increased leukotriene c4 synthesis accompanied enhanced leukotriene c4 synthase expression and activities of ischemia-reperfusion-injured liver in rats.
BACKGROUND
Hepatic ischemia-reperfusion (I/R) injury is an important clinical issue and relates to cysteinyl leukotrienes (LTs), the first committed synthesis step of which is that LTC4 synthesis enzymes including leukotriene C4 synthase (LTC4S), microsomal glutathione-S-transferase (mGST)2, and mGST3-catalyzed LTA4 and reduced glutathione (GSH), to generate LTC4. However, the mechanisms of LTC4 generation during hepatic I/R are far from being elucidated.


METHODS
Adult male Sprague Dawley rats were divided into two groups: sham group (control) and I/R group. Liver was subjected to 60 min of partial hepatic ischemia followed by 5 h of reperfusion; saline was administered intravenously. LTC4 content, the activities, and expressions of LTC4 synthesis enzymes were examined with reversed phase high-performance liquid chromatography, reverse transcriptase-polymerase chain reaction, immunoblot, and immunohistochemistry, respectively. Liver damage was assessed by serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) measurements and histological observation. The superoxide dismutase (SOD) activity and malondialdehyde (MDA) level in liver tissue were used to evaluate lipid peroxidation, and oxidative stress was estimated by the reduced GSH level in liver tissue in the pathological process.


RESULTS
Compared with control, LTC4 content, the LTC4 synthesis enzymes' activities, and the mRNA and protein expressions of LTC4S were significantly increased, while the mRNA expressions of mGST2 and mGST3 were declined obviously in rat liver during I/R (P < 0.05); most hepatocytes and sinusoidal endothelial cells expressed intensively LTC4S in an I/R-sensitive manner. This was accompanied by the increase in serum ALT and AST levels together with liver tissue MDA content (P < 0.05), the decrease in liver tissue GSH level, and SOD activity (P < 0.05), as well as histological damage. There were no differences in the protein expression of mGST3 between control and I/R groups.


CONCLUSIONS
These results demonstrated that hepatic I/R injury up-regulated the mRNA and protein expressions of LTC4S in hepatocytes and sinusoidal endothelial cells and enhanced the activities of the LTC4 synthesis enzymes. It suggests that LTC4 accumulation after hepatic I/R can be caused partially by LTC4S expression up-regulation and the LTC4 synthesis enzymes' activities augment to which LTC4S rather than mGST2 or mGST3 may mainly contribute.
Yang
Shu-Long
SL
Institute of Pharmacology--Toxicology and Biochemical Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, China.
Huang
Xin
X
Chen
Hai-Fei
HF
Xu
Dan
D
Chen
Li-Jun
LJ
Kong
Yin
Y
Lou
Yi-Jia
YJ
D000410:Alanine Transaminase / Q000097:blood; D000818:Animals; D001219:Aspartate Aminotransferases / Q000097:blood; D002851:Chromatography, High Pressure Liquid; D015971:Gene Expression Regulation, Enzymologic; D005982:Glutathione Transferase / Q000235:genetics* / Q000378:metabolism*; D007150:Immunohistochemistry; D017997:Leukotriene C4 / Q000096:biosynthesis* / Q000378:metabolism; D008099:Liver / Q000201:enzymology*; D008297:Male; D008315:Malondialdehyde / Q000378:metabolism; D008862:Microsomes, Liver / Q000201:enzymology; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000378:metabolism / Q000503:physiopathology*; D013482:Superoxide Dismutase / Q000378:metabolism
D012333:RNA, Messenger; D017997:Leukotriene C4; D008315:Malondialdehyde; D013482:Superoxide Dismutase; D005982:Glutathione Transferase; D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase; C041116:leukotriene-C4 synthase