GdCl3 abates hepatic ischemia-reperfusion injury by inhibiting apoptosis in rats.
BACKGROUND
Gadolinium chloride (GdCl(3)) is a specific inhibitor of Kupffer cells (KCs), which are important promoters of various liver injuries. It is therefore of interest to explore the role of KCs in liver ischemia-reperfusion injury and their relations with apoptosis caused by ischemia-reperfusion injury.


METHODS
One hundred male Wistar rats (190-210 g, 6-7 weeks old) were divided into two groups at random, GdCl(3) group and control group. Samples were collected at 0.5, 1, 6, 12, and 24 hours from each group after reperfusion. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured by an automatic biochemical analyzer. TNF-alpha in serum was measured by enzyme-linked immunosorbent assay (ELISA). Malondialdehyde (MDA) in the liver mitochondria was measured by a colorimetric method. Pathological changes in the liver and immunohistochemical staining for caspase-3 were observed under an optical microscope. The ratio of apoptotic cells was measured by TdT-mediated dUTP nick-end labeling (TUNEL), and ultrastructural features of apoptosis were observed with a transmission electron microscope (TEM).


RESULTS
The levels of ALT in the GdCl(3) group were lower than those in the control group after reperfusion for 0.5, 1, 6 and 12 hours (P<0.05); and the levels of AST in the GdCl(3) group were lower than those in the control group after reperfusion for 6 and 12 hours (P<0.05). The levels of TNF-alpha in the GdCl(3) group were lower than those in the control group after reperfusion for each time (P<0.05). The concentrations of MDA after reperfusion in the GdCl(3) group were lower than those in the control group after reperfusion for 6, 12 and 24 hours (P<0.05). After reperfusion for 0.5, 1, 6 and 12 hours, the integral optical density (IOD) of caspase-3-positive cells was lower in the GdCl(3) group than in the control group (P<0.05). After reperfusion for 1, 6, and 12 hours, the IOD of cells stained by TUNEL in the GdCl(3) group was lower than that in the control group (P<0.05).


CONCLUSIONS
GdCl(3) inhibits the activity of ALT, AST and TNF-alpha, decreases the accumulation of MDA in mitochondria, and depresses the expression of caspase-3 in liver after ischemia-reperfusion. This may be an important protective mechanism by depressing KCs and indirectly inhibiting liver cell apoptosis.
Li
Jian-Yi
JY
Department of General Surgery, Shengjing Hospital, China Medical University, Shenyang 110004, China.
Gu
Xi
X
Zhang
Wen-Hai
WH
Jia
Shi
S
Zhou
Yong
Y
D000410:Alanine Transaminase / Q000097:blood; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology* / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects*; D001219:Aspartate Aminotransferases / Q000097:blood; D053148:Caspase 3 / Q000378:metabolism; D005682:Gadolinium / Q000494:pharmacology* / Q000627:therapeutic use*; D007728:Kupffer Cells / Q000187:drug effects / Q000473:pathology; D008099:Liver / Q000293:injuries* / Q000378:metabolism / Q000473:pathology; D008297:Male; D008315:Malondialdehyde / Q000378:metabolism; D008930:Mitochondria, Liver / Q000378:metabolism; D023421:Models, Animal; D051381:Rats; D017208:Rats, Wistar; D015427:Reperfusion Injury / Q000097:blood / Q000517:prevention & control*; D014409:Tumor Necrosis Factor-alpha / Q000097:blood
D000893:Anti-Inflammatory Agents; D014409:Tumor Necrosis Factor-alpha; D008315:Malondialdehyde; D005682:Gadolinium; D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase; D053148:Caspase 3; C038958:gadolinium chloride