OBJECTIVE@#To clarify the preparation
methods of four
rat models of
liver ischemia/
reperfusion injury (IRI) and to determine a
liver IRI
animal model that is consistent with clinical conditions, has stable pathological and physiological
injury, and is easy to operate.@*
METHODS@#A total of 160
male Sprague-Dawley (SD)
rats were randomly divided into four groups using an interval grouping
method:
70% IRI (group A), 100% IRI (group B), 70% IRI with 30%
hepatectomy (group C), and 100% IRI with 30%
hepatectomy (group D), with 40
rats in each group. Each model was further divided into sham operation group (S group) and
ischemia groups of 30, 60, and 90 minutes, with 10
rats in each group. After
surgery, the
survival status and awakening
time of the
rats were observed, and the
liver lobectomy weight,
bleeding volume, and
hemostasis time of groups C and D were recorded.
Blood samples were collected by cardiac
puncture after 6 hours of
reperfusion for
determination the levels of
aspartate aminotransferase (AST),
alanine aminotransferase (ALT),
alkaline phosphatase (ALP),
blood urea nitrogen (
BUN),
serum creatinine (SCr), and γ-
glutamyl transpeptidase (γ-GT) in the
serum to assess
liver and
kidney function.
Hematoxylin-
eosin (HE)
staining and immunohistochemical
staining of
macrophages were performed to analyze the
liver tissue structure damage from a pathological perspective.@*RESULTS@#
Rats in group A exhibited earlier awakening and acceptable mental status, while
rats in the other groups showed delayed awakening and poor mental status. The
hemostasis time in group D was approximately 1 second longer than that in group C. The
mortality of
rats subjected to 60 minutes of 70% hepatic
ischemia was 0. Compared to the sham operation group,
rats in each experimental group showed significant increases in
serum levels of AST, ALT, ALP,
BUN, SCr, and γ-GT, indicating impaired
liver and
kidney function in the
rat models of
liver IRI. In groups A, B, and C, the 90-minute
ischemia subgroup exhibited more pronounced elevation in AST, ALT, ALP,
BUN, SCr, and γ-GT levels compared to the 30-minute
ischemia subgroup [AST (U/L) group A, 834.94±56.73 vs. 258.74±18.33; group B, 547.63±217.40 vs. 277.67±57.92; group C, 930.38±75.48 vs. 640.51±194.20; ALT (U/L) group A, 346.78±25.47 vs. 156.58±13.25; group B, 408.40±138.25 vs. 196.80±58.60; group C, 596.41±193.32 vs. 173.76±72.43; ALP (U/L) group A, 431.21±34.30 vs. 315.95±15.64; group B, 525.88±62.13 vs. 215.63±17.31; group C, 487.53±112.37 vs. 272.46±92.33;
BUN (U/L) group A, 18.35±5.63 vs. 14.32±2.30; group B, 30.21±4.55 vs. 17.41±8.14; group C, 20.50±3.64 vs. 15.93±3.22; SCr (U/L) group A, 27.47±8.91 vs. 22.37±5.66; group B, 43.60±15.57 vs. 36.80±7.95; group C, 63.81±20.24 vs. 42.47±7.03; γ-GT (U/L) group A, 15.64±3.57 vs. 6.82±1.48; group B, 9.28±1.91 vs. 5.62±1.21; group C, 10.98±3.18 vs. 5.67±1.10; all P < 0.05]. The 100% IRI 90-minute group and 100% IRI 90-minute group with 30%
hepatectomy exhibited more pronounced increases in the above-mentioned
indicators compared to the corresponding 70% IRI
control group, indicating increased
liver and
kidney damage in
rats subjected to combined
blood flow occlusion and
hepatectomy. HE
staining showed clear
liver tissue structure with intact and orderly arranged
cells in the sham operation group, while the experimental groups exhibited
cell structure damage, including
cell rupture or collapse,
cell swelling, nuclear pyknosis, deep
cytoplasm staining,
cell shedding, and
necrosis. The interstitium showed infiltration of inflammatory
cells. Immunohistochemical
staining revealed a higher number of
macrophages in the experimental groups compared to the sham operation group.@*CONCLUSIONS@#Four models of
liver IRI in
rat were successfully established. As the duration and severity of hepatic
ischemia increased,
liver cell ischemia worsened, leading to increased hepatocellular
necrosis and exhibiting characteristic features of
liver IRI. These models can effectively simulate
liver IRI following
liver trauma, with the group subjected to 100%
ischemia and 30%
hepatectomy showing the most severe
liver injury. The designed models are reasonable, easy to perform, and exhibit good reproducibility. They can be used for investigating the mechanisms,
therapeutic efficacy, and diagnostic
methods related to clinical
liver IRI.