| Objective:To observe the effect of traditional Chinese medicines(TCMs)on liver regeneration,gastrointestinal motility and barrier function in rats after partial hepatectomy.Methods:Healthy male Sprague-Dawley rats were randomly divided into two groups:hepatic fibrosis group and normal group,48 rats in each group.The liver fibrosis group was subcutaneously injected with 40%carbon tetrachloride-olive oil mixture at 5 ml/kg twice a week for 10 weeks to establish liver fibrosis model,while the normal group was subcutaneously injected with normal saline at the same dose.In the fibrosis group,blood was collected from caudal vein at 2,6 and 10 weeks during the modeling process,and serum was collected after centrifugation to detect the levels of transaminase,bilirubin,hyaluronic acid(HA)and laminin(LN).After successful modeling,rats with hepatic fibrosis and normal rats were divided into four groups:control group(C)without surgical treatment;sham group(S)with laparotomy instruments to free the posterior closure of the perihepatic ligament;partial hepatectomy group(PH)with laparotomy to free the perihepatic ligament,resection of the left and middle lobes of the liver;Medicines group(M)was treated with TCMs during perioperative period on the basis of PH group.HE staining,Masson staining and immunohistochemistry were used to detect the expression of alpha-smooth muscle actin(α-SMA)in the liver,and to evaluate the degree of liver fibrosis.The rats in M group were administered with TCMs from 1 day before operation to 8 o’clock in the morning and evening on the 2nd or 6th day after operation.The rats in other groups were administered with normal saline at the same dose.The general condition,serum transaminase,bilirubin,hepatocyte growth factor(HGF),motilin level(MTL),liver regeneration rate,gastrointestinal motility and mucosal barrier function were observed.During the experiment,the activity,spirit,diet and urine of each group were recorded;blood was taken from caudal vein 2,4,7 days after operation,serum was collected after centrifugation,and stored at-80℃ for examination.Six rats in each group were executed on the 3rd and 7th day after operation.Liver samples were taken to measure the weight of liver to evaluate the liver regeneration rate.The expression levels of proliferating cell nuclear antigen(PCNA)and Ki-67 in liver were detected by immunohistochemistry to evaluate the effect of TCMs on liver regeneration ability after PH in rats.The intestinal propulsion test was carried out by activated carbon gel method.Serum MTL level was determined by ELISA method,and the influence of TCMs on gastrointestinal motility was evaluated.Intestinal specimens were collected,HE staining and intestinal villus height were measured,and the location and expression of mucoprotein-2(MUC-2)were detected by immunohistochemistry.The effects of TCMs on intestinal barrier function in rats were evaluated.Results:1.Liver fibrosis model:The levels of serum AST,ALT,TBIL,HA and LN increased progressively during 2,6 and 10 weeks of the modeling process;HE and Masson staining showed the destruction and disorder of hepatic lobule structure,a large amount of collagen fibers deposited and pseudolobule formation;immunohistochemical staining of alpha-SMA showed that the expression level of alpha-SMA in fibrotic liver increased significantly,indicating that the model had been successfully established.2.Liver function recovery after PH:After liver fibrosis and normal operation,the levels of AST,ALT and TBIL in serum of PH group and M group were significantly higher than those of C group;On the second day after operation,there was no significant difference in the levels of AST,ALT and TBIL between PH group and M group(P>0.05);on the fourth and seventh day after operation,the levels of AST,ALT and TBIL in serum of M group were significantly improved compared with those of PH group(P<0.05).3.Liver regeneration ability:In hepatic fibrosis rats,the liver regeneration rate of M group was significantly higher than that of PH group on the 3rd and 7th day after operation(P<0.05);in normal rats,there was no significant difference in the liver regeneration rate between PH group and M group on the 3rd day after operation(P>0.05).On the 7th day after operation,the liver regeneration rate of M group was significantly higher than that of PH group,with statistical significance(P<0.05).The rate of liver regeneration in normal rats was significantly higher than that in liver fibrosis rats(P<0.05).On the 2nd,4th and 7th day after operation,HGF levels in PH and M groups were significantly higher than those in C and S groups(P<0.05),while those in M group were significantly higher than those in PH group(P<0.05).The positive rates of PCNA and Ki-67 in hepatocytes of M group were significantly higher than those of PH group on the 3rd and 7th day after operation in hepatic fibrosis and normal rats(P<0.05).The positive rates of PCNA and Ki-67 in hepatocytes of normal rats after PH operation were significantly higher than those of hepatic fibrosis rats(P<0.05).4.Restoration of gastrointestinal motor function:Activated carbon intestinal propulsion index in M group was significantly higher than that in group PH on the 3rd and 7th day after operation in rats with hepatic fibrosis(P<0.05);in addition,compared with normal rats,activated carbon intestinal propulsion index in rats with hepatic fibrosis was significantly lower(P<0.01).The serum MTL levels in M group were significantly higher than those in PH group at the 2nd,4th and 7th day after liver fibrosis and normal operation(P<0.05).5.Effect on intestinal mucosal barrier:HE staining results showed that the intestinal villus height of rats in M group was significantly higher than that of rats in PH group at 3 and 7 days after operation(P<0.05);compared with normal rats,the intestinal villus height of rats with hepatic fibrosis was significantly lower(P<0.05).The expression of MUC-2 in intestinal mucosa of M group was higher than that of PH group on the 3rd and 7th day after operation in liver fibrosis and normal rats(P<0.05).Conclusion:The application of TCMs in perioperative period of PH can improve the liver regeneration ability of normal and hepatic fibrosis rats.At the same time,it can also promote the recovery of gastrointestinal motility and protect the intestinal mucosal barrier function after operation. |
PDF Full Text Request