| Objectives:To establish the model of cholestatic hepatocyte damage in rats by common bile duct ligation and build a randomized controlled trial, in which experimental rats were given the extracts from Penthorum chinense Pursh or ursodeoxycholic acid by gavage. To observe the changes of liver function and lipid peroxidation index by serological tests. The histopathology and ultrastructure of hepatocytes in the model animals were observed by light microscopy and transmission electron microscopy. The protective effects of the extracts from Penthorum chinense Pursh for cholestatic hepatocyte damage were investigated and the mechanism was discussed through lipid peroxidation of oxidative stress pathway.Methods: 1.Establishing cholestatic hepatocyte damage rat models:Sprague-Dawley rats were raised in the SPF asepsis animal room and were adapted to the environment after 7 days. 2% pentobarbital(100mg/kg) was intraperitoneally injected to anesthetize the rat. The common bile duct was revealed through laparotomy. The common bile duct was double ligated and transected between the two lines.2.Groups: 15 normal male rats were randomly selected as control group(group 1).Total 45 male SD rats with successful modeling were randomly divided into three groups: common bile duct ligation group(group 2), the extracts from Penthorum chinese Pursh group (group 3), ursodeoxycholic acid group(group 4).every group includes 15 rats.3.At postoperative 2d, experimental rats were given medicines orally(4 m L, 1 time/day, for four weeks). Group 1 and group 2 were given equal dose of normal saline orally. Group 3 were given the extracts from Penthorum chinense Pursh orally,(4 000mg·kg-1). Group 4 were given ursodeoxycholic acid orally,(30mg ·kg-1).4.The rats were anaesthetized at preoperative 30 min, 7th day, 14 th day after the operation to take the blood from heart for liver function test. AST,ALT,ALB,TBIL,DBIL,ALP were tested. The rats were anaesthetized at 7th day and 14 th day after the operation; partial hepatic tissues of rats were taken for biopsy. If the rat died in the experimental process,An autopsy should be done in time. At 7th day and 14 th day after operation, biopsy was performed for pathologic exam. Histopathology and ultrastructure of liver tissue in rats were observed by light microscopy and transmission electron microscopy. Meanwhile, the indexes of MDA, ROS,SOD were tested.Results: 1. The liver function test results: At postoperative 3 d, 7 d, 14 d, AST, ALT, TBIL, DBIL, ALP of group 2 were significantly higher than that of group 1(P < 0.05). Compared with group 2, AST, ALT, TBIL, DBIL, ALP of group 3, 4 decreased obviously(P < 0.05). TBIL, DBIL of group 3 at postoperative 7 d, 14 d were lower than those of group 4(P < 0.05). Propagated in group 1, group 2, group 3, 4 at 3 d, 7 d, and 14 d after operation had no significant difference(P > 0.05).2. pathological results: The pathology of liver tissue in group 1 at 7 d, 14 d postoperative had no obvious change: the structure of hepatic lobule was complete, long axis of the hepatic lobules central contorts the central vein, liver blood sinus and rope around the central vein, radiate out in neat cells, liver cells line showed obvious radial, portal area without deformation, necrosis and inflammation cells infiltration. Group 2 at postoperative 7 d: lobular structure was incomplete, unclear profile, liver cell flated in shape, bile duct epithelial cells swelling, necrosis, portal area with inflammatory cells infiltration, liver cells markedly swollened, part a cavitation degeneration. Group 2 at postoperative 14 d: lobular structure chaos, central vein, portal area expansion deformation, a large number of inflammatory cells infiltration, liver cell swelling, large area, showed cavitation model change, collagen deposition around the central vein and portal area. Compared with group 1, the damage of liver tissue in group 1 was serious. Group 3, 4 at postoperative 7 d, 14 d: lobular structure was complete, the basic normal interlobular bile duct, portal area had a small amount of inflammatory cells infiltration, normal liver cells form, some swelling, individual assumes the cavitation degeneration. Liver tissue damage degree in group 3,4 was a significant reduction than that in group 2. 3. Electron microscopic pathology results: group 1 at postoperative 7 d: adjacent liver cells arranged neatly, dividing line was clear, structure was normal, large, round nucleus, clear and obvious nucleoli, chromatin distribution was normal, the nuclear membrane structure was clear, the mitochondria structure was normal, ridge structure was clear, structure was clear, endoplasmic reticulum arrangement rule, in the cytoplasm were rich in glycogen, dyeing uniformity. The liver cell ultrastructure in group 1 at postoperative 14 d was similar as that at postoperative 7 d. Group 2 at postoperative 7 d: part of the cells was edema, intracellular lipid droplets in size, the nucleus edge together, nuclear, heterochromatin increased, nuclear membrane shrivel, mitochondria swelling and part of the crest fracture, nuclear weeks gap widened. Group 2 at postoperative14 d: liver cell edema, intracellular lipid droplets in great quantities, the nucleus presented edge set signs, nuclear membrane shrivel, markedly swollen mitochondria, part of the crest fracture, show cavitations model change. Group 3, 4 at postoperative 7 d, and 14 d: basic normal cellular structure, slightly swelling mitochondria, endoplasmic reticulum expansion, to a lesser damage. Liver cell ultrastructure had no significant difference between two groups. 4. Lipid peroxidation index results: MDA, ROS in group 2 at postoperative 7d, 14 d were significantly higher than that in group 1 separately(P < 0.05). MDA and ROS in group 3, 4 at postoperative 7d, 14 d were lower than that in group 2(P < 0.05), MDA and ROS had no significant difference between group 3 and group 4(P > 0.05). SOD in group 2 at postoperative 7 d, 14 d decreased which were significantly lower than that in group 1(P < 0.05). SOD in group 3, 4 at postoperative 7 d, 14 d were higher than that in group 2(P < 0.05), SOD had no obvious difference between group 3 and group 4(P > 0.05).Conclusion: 1. The extracts from Penthorum chinense Pursh can reduce liver serum enzyme index in cholestasis rat liver tissue, reduce cholestatic jaundice, maintain mitochondrial morphological structure and function, reduce oxidative stress reaction of cell damage and protect liver cell damage of cholestasis. 2. The mechanism may be related to relieve lipid peroxidation damage caused by oxidative stress. |
PDF Full Text Request