Protective Effects Of Drug 0412 On Chronic Hepatic Injury In Rats

ObjectiveTo study the protective effects of drug 0412 on carbon tetrachloride induced chronic hepatic injury in male Wister rats by observing the change of pathological histology, hepatic function indexes and hepatic fibrosis indexes, and as well as test its pharmacological effects and possible mechanism;meanwhile, to observe the spontaneous reverse of chronic hepatic injury in rats.Materials and methodsChronic hepatic injury was induced by intraperitoneal injection of 40% carbon tetrachloride (CCl₄) with 0.3ml/100g weight (0.2ml/100g weight from the third week) twice a week. At the end of the fourth week, those model rats were divided into five groups: model control group (CCl₄-injured), 0412 high dose group (CCL₄-injured+0412 , 100mg/kg/day), medium dose group (CCl₄-injured+0412, 50mg/kg/day)' and low dose group (CCl₄-injured+0412, 25mg/kg/day) , CCl₄ ceased group, in addition, a normal control group was set up. From the fifth week, model group was still treated with CCl₄ as before, three 0412 treated groups were respectively treated with intragastric administration of drug 0412 with 100mg/kg, 50mg/kg or 25mg/kg everyday atthe base of CC14 treating as before, CC14 ceased group was treated with the same dose of normal saline at the base of ceasing CC14. During the experiment, all the rats' general status including mental status, activities and excretion were observed. After 20 days' therapy, all the rats were weighed and killed after 12 hours' depriving of food or water. Then, outlook, color, texture of liver were observed and the liver index ( liver weight per body weight) was calculated;Alanine transaminase(ALT), aspartate aminotransferase(AST), albumin(ALB) and albumin/globulin(A/G) in serum, serum hyaluronic acid (HA), hydroxyproline (Hyp) in liver were measured. Besides, we also observed the changes of liver histopathology and expression of a -smooth muscle actin (a -SMA) by immunohistochemistry method.Results1. From the start of treating to the end, there were 6 rats died in the model group, each 4 rats died in the three 0412 treated groups and 2 died in the CCL ceased group. During the experiment, rats in the model group showed depressed, anorexia and retardation. All the phenomenons would extinction in 12-24 hours, besides, rats in the model group lost weight markedly and suffered diarrhea. While general status of the 0412 treated groups was much better than the model group in the late period. The general status of CC14 ceased group was much better than the model group after stopping CCU treating.2. Normal rat liver capsules were smooth, and their surfaces were red brown with soft texture. In the model group, liver adhered to other tissues seriously and capsules were rough;its colors was khaki and edge was blunt with hard texture;the liver index was much higher than the normal group (K0. 01). While in the 0412 treated groups, liver lobules also adhered to each other, but its color was wine and the texture was softer than themodel group, liver index was much lower than the model group (KO. 05). In the CC14 ceased group, the liver index was close to the normal group, and the liver was wine with medium texture.3. Compared to the normal group, ALT, AST of model group markedly increased (R0. 01);ALB, A/G decreased (K0. 01). Liver function indexes of three 0412 treated groups were close to the normal group, but compared to the model group, their ALT, AST markedly decreased (K0. 01), the high and medium dose groups' ALB markedly increased (K0. 05), all groups' A/G increased (K0. 01). ALT, AST levels of the CC14 ceased group had no statistical difference with the normal group, but were obviously lower than the model group (K0. 01).4. Compared to the normal group, serum HA of model group markedly increased (KO. 01). Serum HA of-three 0412 treated groups was close to the normal group but was much lower than the model group (FKO. 05). Serum HA of the CCI4ceased group was a little higher than the normal group (P>0. 05) but much lower than the model group (K0. 01).5. Compared to the normal group, liver Hyp of model group markedly increased (/K0. 01). Liver Hyp of three 0412 treated groups had no statisticaldifference with the normal group but was much lower than the model group (KO. 01). Liver Hyp of the CC14 ceased group was a little higher than thenormal group (P>0. 05) but was obviously lower than the model group (K0. 05).6. HE and Sirius red staining showed: in the normal group, there was no infiltration of inflammation cells or fibrotic Hyperplasia in liver, with clear lobule structure and orderly hepatic cord;in the model group, there was extensive vacuole-like hepatic steatosis and focal necrosis with infiltration of inflammation cells, meanwhile the bunchy collagen fibre proliferated obviously from the central venous to the liver lobules,forming fibrotic septa;While in 0412 treated groups, the pathological changes of steatosis and fibrosis were obviously alleviated and there was no infiltration of inflammation cells with clear lobule structure;Pathological change of the CC14 ceased group was close to the normal group, there was only light fibrosis between hepatic sinuses or lobules with no 'infiltration of inflammation cells or steatosis.7. a -SMA only expresses in vascular smooth muscle cell in the normal, 0412 treated and CC14 ceased groups, while still expresses in the portal and walls of hepatic sinuses in the model group.ConclusionDrug 0412 can effectively protect the CC14 induced chronic liver injury in rats by improving the hepatic function, change of pathological histology and inhibiting hepatic fibrosis. Besides, indexes of the spontaneous model of chronic hepatic injury are all close to the normal group. That shows wiping off the causes of disease plays an important role in the reverse of chronic hepatic injury especially the hepatic fibrosis.