Establishment of a hepatic cirrhosis and portal hypertension model by hepatic arterial perfusion with 80% alcohol

Abstract

Aim: To determine the feasibility and safety of establishing a porcine hepatic cirrhosis and portal hypertension model by hepatic arterial perfusion with 80% alcohol.

Methods: Twenty-one healthy Guizhou miniature pigs were randomly divided into three experimental groups and three control groups. The pigs in the three experimental groups were subjected to hepatic arterial perfusion with 7, 12 and 17 mL of 80% alcohol, respectively, while those in the three control groups underwent hepatic arterial perfusion with 7, 12 and 17 mL of saline, respectively. Hepatic arteriography and direct portal phlebography were performed on all animals before and after perfusion, and the portal venous pressure and diameter were measured before perfusion, immediately after perfusion, and at 2, 4 and 6 wk after perfusion. The following procedures were performed at different time points: routine blood sampling, blood biochemistry, blood coagulation and blood ammonia tests before surgery, and at 2, 4 and 6 wk after surgery; hepatic biopsy before surgery, within 6 h after surgery, and at 1, 2, 3, 4 and 5 wk after surgery; abdominal enhanced computed tomography examination before surgery and at 6 wk after surgery; autopsy and multi-point sampling of various liver lobes for histological examination at 6 wk after surgery.

Results: In experimental group 1, different degrees of hepatic fibrosis were observed, and one pig developed hepatic cirrhosis. In experimental group 2, there were cases of hepatic cirrhosis, different degrees of increased portal venous pressure, and intrahepatic portal venous bypass, but neither extrahepatic portal-systemic bypass circulation nor death occurred. In experimental group 3, two animals died and three animals developed hepatic cirrhosis, and different degrees of increased portal venous pressure and intrahepatic portal venous bypass were also observed, but there was no extrahepatic portal-systemic bypass circulation.

Conclusion: It is feasible to establish an animal model of hepatic cirrhosis and portal hypertension by hepatic arterial perfusion with 80% alcohol, however, the safety of this model depends on a suitable perfusion dose.

Keywords: Alcohol; Animal model; Hepatic arterial perfusion; Hepatic cirrhosis; Portal hypertension.

Figure 1

Hepatic artery variation. A: Hepatic arteriography of experimental group 2 before perfusion showed good distribution of hepatic arteries and their branches; B: After hepatic arterial perfusion in experimental group 2, small peripheral branches of hepatic arteries were significantly reduced compared with before perfusion.

Figure 2

Portal vein variation. A: Direct portal phlebography of experimental group 2 before perfusion showed the portal venous branches clearly; B: Portal phlebography immediately after perfusion showed a significant reduction in small portal venous branches and a widened main portal vein; C: Portal phlebography at 6 wk after perfusion showed that a large number of irregular bypass branches of intrahepatic portal veins were formed, which communicated directly with hepatic veins, and the main portal vein was still wider than that before perfusion.

Figure 3

Liver size variation. A: Computed tomography of experimental group 2 before perfusion demonstrating normal liver size; B: At 6 wk after perfusion, liver size was markedly increased.

Figure 4

Spleen size variation. A: Computed tomography of experimental group 2 before perfusion demonstrating normal spleen size and the spleen was thin and curled; B: At 6 wk after perfusion, spleen size was markedly increased; the spleen was not curled and extended into the hepatic fissure.

Figure 5

Pathological variation. A: Within 6 h after perfusion in experimental group 2, thrombosis was evident in interlobular veins; B: At 2 wk after perfusion in experimental group 2, irregular, massive hepatocyte degeneration and necrosis were observed; C: At 4 wk after perfusion in experimental group 2, bile duct proliferation and dilation were evident; D: At 6 wk after perfusion in experimental group 2, pseudo-lobules formed with fibrous tissue proliferation and bridging fibrosis. Hematoxylin-eosin staining, magnification × 40 (A); × 200 (B-D).

Figure 6

Gross sample of cirrhotic liver. A, B: At 6 wk after perfusion in experimental group 2, gross samples of diaphragmatic and ventral liver showed that hepatic tissues were hard and had poor elasticity, and various liver lobes were enlarged; C: Local amplification of the gross sample showed that there were relatively uniform regenerative nodules on the liver surface.

Figure 7

Gross sample of the spleen (A) and stomach (B). A: At 6 wk after perfusion in experimental group 2, the gross sample of the spleen showed that the spleen was hard and congestive, and had a non-uniform color; B: At 6 wk after perfusion in experimental group 2, the gross sample of the stomach showed that there were small varicose veins on the stomach surface.