OBJECTIVETo determine the effect of pneumoperitoneum on liver indocyanine green (ICG) metabolism and the hepatic blood flow in normal and cirrhotic rats.

METHODSThirty male Wistar rats were randomized into five groups: normal anaesthesia group, normal laparotomy group, normal pneumoperitoneum group, cirrhosis + anaesthesia group, and cirrhosis + pneumoperitoneum group. Liver cirrhosis was induced in two groups by injecting carbon tetrachloride subcutaneously plus drinking 5% alcohol. ICG clearance tests were performed in all the rats.

RESULTSThe ICG level in the normal laparotomy group (0.662 micro g/ml) was higher than that in the normal anesthesia group (0.645 micro g/ml), but the difference was not significant (P > 0,05). The ICG level in the normal pneumoperitoneum group (0.967 micro g/ml) was significantly higher than that in the normal anesthesia and normal laparotomy groups (P < 0.05). The ICG levels in two cirrhotic groups were significantly higher than those in the other three groups (P < 0.05). The ICG level in the cirrhosis + pneumoperitoneum (1.348 micro g/ml) was significantly higher than that in the cirrhosis + anesthesia group (1.198 micro g/ml) (P < 0.05).

CONCLUSIONSDuring laparoscopic surgery, pneumoperitoneum could decrease the liver ICG clearance rate and the hepatic blood flow, which are of clinical significance in determining the state of liver cirrhotic.

Animals ; Disease Models, Animal ; Indocyanine Green ; metabolism ; Liver Circulation ; Liver Cirrhosis, Experimental ; physiopathology ; Male ; Pneumoperitoneum, Artificial ; Rats ; Rats, Wistar

To investigate the role of heme oxygenase(HO), a catalyzing enzyme of heme to produce CO, in modulation of systemic circulation in CCl4-induced cirrhotic rats. Saline(vehicle) and ZnPP were s.c. injected into the posterior necks of rats respectively and the rats were then anesthetized by pentobarbital sodium in four hours. Mean arterial pressure (MAP, kPa), heart rate (HR, b/min) and portal pressure (PP, cm/H2O) were measured by indwelling catheter. Plasma CO was determined by Chalmers method. Heme oxygenase acivity was determined by the rate of bilirubin formation. The cirrhotic rats showed significant hyperdynamic circulation indicated by decreased mean arterial pressure [MAP, (15.6+/-1.7) vs (18.9+/-0.9) kPa, t = 4.52, P less than 0.01] and increased portal pressure [PP, (16.7+/-0.8) vs (8.8+/-0.3) cm H2O, t = 23.10, P less than 0.01] as compared to normal control rats(NS). ZnPP could cause a significant increase in MAP [(17.3+/-1.5) vs (15.6+/-1.7) kPa, t = 2.18, P less than 0.05] and significant decrease in PP [(13.2+/-0.7) vs (16.7+/-0.8) cm H2O, t = 8.53, P less than 0.01] in cirrhotic rats. The cirrhotic group presented a significant increase in plasma CO [(18.0+/-1.9) vs (10.4+/-1.3)mumol/L, t = 8.42, P less than 0.01] and HO activity in the spleens [(11.1+/-0.9) vs (6.5+/-0.9) nmol bilirubin/mg protein/h, t = 9.28, P less than 0.01] and intestines [(2.5+/-0.1) vs. (1.3+/-0.2) nmol bilirubin/mg protein/h, t = 15.1, P less than 0.01]. ZnPP could cause significant decreases in plasma CO and HO activity in liver, spleen and intestine of both control and cirrhotic rats. HO-CO system activation may be an important reason for the hemodynamic disturbance of liver cirrhosis.
Animals ; Carbon Monoxide ; metabolism ; Heme Oxygenase (Decyclizing) ; metabolism ; Hemodynamics ; Liver ; physiopathology ; Liver Cirrhosis, Experimental ; metabolism ; physiopathology ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo noninvasively evaluate hepatic functional blood flow, intrahepatic shunt rate and hepatic functional reserve in both normal and cirrhotic liver using D-sorbitol and indocyanine green measured by high performance liquid chromatography (HPLC).

METHODSMale Sprague-Dawley (SD) rats were divided into normal control and cirrhotic group in which the rats were administrated with tetrachloride. Then the isolated perfused liver models were established. The pharmacokinetic indexes of D-sorbitol and indocyanine green (ICG) were measured by the traditional spectrophotometry (SPEC) and HPLC respectively.

RESULTS(1) HPLC showed that ICG contained genuine ICG (ICGg) and ICG degraded products (ICGdp), which had similar spectrum but metabolic kinetics different with the retention time of 8.9 minutes and 24.2 minutes respectively. (2) Hepatic intrinsic metabolic capacity (QINT, I) was (36.57+/-13.03) ml/min in control group and (14.39+/-5.13) ml/min in cirrhotic group (t=7.08, P<0.01). (3) Hepatic functional blood flow (QFUNC) in cirrhotic group declined, compared with that in control group (34.06 ml/min+/-5.12 ml/min vs. 17.54 ml/min+/-7.02 ml/min, t=8.41, P<0.01), while intrahepatic shunt rate (QIHS) increased markedly (9.9%+/-1.4% vs. 47.5%+/-20.9%, t=8.35, P<0.01).

CONCLUSION(1) HPLC method is superior to SPEC in measuring ICG, because it can avoid the disturbance from ICGdp, so that ICG measured by HPLC is valid for QINT, I evaluation. (2) The hepatic clearance of D-sorbitol measurement is a noninvasive and reliable method for evaluating the total blood flow in normal liver, and hepatic functional blood flow and intrahepatic shunt rate in cirrhotic liver. (3) Combining D-sorbitol with indocyanine green measurement is helpful for assessment of liver functional reserve.

Animals ; Carbon Tetrachloride Poisoning ; physiopathology ; Indicators and Reagents ; pharmacokinetics ; Indocyanine Green ; pharmacokinetics ; Liver ; physiopathology ; Liver Cirrhosis, Experimental ; chemically induced ; metabolism ; physiopathology ; Male ; Rats ; Rats, Sprague-Dawley ; Sorbitol ; pharmacokinetics

Animals ; Liver ; pathology ; Liver Cirrhosis, Experimental ; metabolism ; pathology ; physiopathology ; Male ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptor, Platelet-Derived Growth Factor beta ; metabolism

Animals ; Blood Pressure ; physiology ; Hemodynamics ; physiology ; Hypertension, Portal ; blood ; metabolism ; physiopathology ; Liver Cirrhosis, Experimental ; blood ; metabolism ; physiopathology ; Male ; Neuropeptide Y ; blood ; metabolism ; Portal Pressure ; physiology ; Rats ; Rats, Sprague-Dawley ; Sodium ; urine

BACKGROUND: This study was designed to determine the relationship of propranolol pharmacokinetic parameters with portosystemic shunt in CCl4-induced cirrhotic rats. METHODS: Cirrhotic rats(n=6) were induced by intramuscular injection of CCl4 in olive oil(two time per weeks) for 12 weeks. Controls (n=6) were injected intramuscularly with the same dose of olive oil for 12 weeks. We evaluated the amount of portosystemic shunt by thallium-201 per rectal scintigraphy. After intravenous bolus injection of propranolol (2mg/kg) to rats, the serum propranolol concentrations were analyzed by a HPLC-fluorimetric detector system. Pharmacokinetic parameters such as C0, AUC, t(1/2(beta)), and CLp were determined in each group. Then, a small amount of heptic tissue was obtained and subjected to determination of the hepatic collagen content by quantitating 4-hydroxyproline and were inspected by microscope after hematoxylin and eosin stain. RESULTS: In liver biopsy, liver fibrosis progressed in CCl4-induced cirrhotic rats. The serum concentrations of propranolol were significantly (p < 0.01) elevated in CCl4-induced cirrhotic rats. Mean amount of 4-hydroxyproline, mean H/L ratio, and mean AUC in CCl4-induced cirrhotic rats was significantly (p < 0.01) higher than that in control rats. There was a relationship between AUC, H/L ratio, and amount of 4-hydroxyproline. CONCLUSION: H/L ratio may help in the selection of drug dosage (especially blood flow dependent drug) in pre-clinical studies for chronic liver disease during the drug development process.
Animals ; Carbon Tetrachloride Poisoning/*complications ; Chromatography, High Pressure Liquid ; English Abstract ; Liver Cirrhosis, Experimental/*metabolism/physiopathology ; Portal System/physiopathology ; Propranolol/*pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Thallium Radioisotopes/diagnostic use

BACKGROUND/AIMS: The Wnt/beta-catenin signaling pathway has been reported to play an important role in liver fibrosis. This study was designed to investigate whether mesoderm-specific transcript homologue (Mest), a strong negative regulator of Wnt/beta-catenin signaling, could inhibit liver fibrosis. METHODS: pcDNA-Mest was transfected into hepatic stellate cells (HSCs) and rats. Rats were randomly divided into four groups: normal group (normal saline), treatment group (pcDNA-Mest+CCl4), control group (pcDNA-neo+CCl4), and model group (normal saline+CCl4). Changes in liver pathology were evaluated by hematoxylin and eosin and Masson's trichrome staining. The levels of alanine transaminase, aspartate transaminase, lactic dehygrogenase, hyaluronic acid, and laminin in the serum and hydroxyproline in the liver were detected by biochemical examination and radioimmunoassay, respectively. The expression and distribution of beta-catenin, alpha-smooth muscle actin (alpha-SMA), Smad3, and tissue inhibitor of metalloproteinase type I were determined, and the viability of the HSCs was tested. RESULTS: Our data demonstrate that Mest alleviated CCl4-induced collagen deposition in liver tissue and improved the condition of the liver in rats. Mest also significantly reduced the expression and distribution of beta-catenin, alpha-SMA and Smad3 both in vivo and in vitro, in addition to the viability of HSCs in vitro. CONCLUSIONS: We found that Mest attenuates liver fibrosis by repressing beta-catenin expression, which provides a new therapeutic approach for treating liver fibrosis.
Animals ; Carbon Tetrachloride/toxicity ; Cells, Cultured ; Hepatic Stellate Cells/physiology ; Liver Cirrhosis, Experimental/*physiopathology ; Male ; Proteins/*physiology ; Random Allocation ; Rats, Wistar ; Transfection ; Wnt Signaling Pathway/*physiology ; beta Catenin/metabolism

OBJECTIVETo explore the mechanism of Danggui Buxue Decoction (, DBD) on the liver fifibrosis related to hepatic lipid peroxidation and matrix metalloproteinases (MMP) -2/9 activities.

METHODSThe liver fifibrosis in 28 rats was induced by an injection of carbon tetrachloride (CCl(4)) and fed with high lipid and low protein diet for 6 weeks, the model rats were randomly divided into the model group and DBD treated group, 14 in each group, and another 10 rats as the normal group were observed as well. Rats in the DBD group were administered with DBD at the dose of 6 g/kg body weight for 6 weeks since CCl(4) intoxication. The hepatic inflammation and fibrosis were examined with HE and Sirius red stain. The liver function including serum alanine aminotransamine (ALT), aspartate transamine (AST), albumin (Alb) and total bilirubin (TBIL), liver triglyceride (TG) and malondialdehyde (MDA) contents, superoxide dismutase (SOD) activity were assayed. Hepatic hydroxyproline (Hyp) content was detected with Jamall's method. The alpha-SMA expression was analyzed by immunohistochemistry and the Western blot. Liver MMP-2 mRNA was analyzed with Real-time PCR, and MMP-2/9 activities were measured with gelatin zymography and in situ zymography.

RESULTSCompared with the normal group, the levels of ALT, AST and TBIL, the content of Hyp, TG and MDA were remarkably increased, the Alb content and SOD activity were signifificantly decreased in the model group (P<0.05), and higher levels of MMP-2 mRNA and MMP-2/9 activities (P<0.01), the hepatic fatty degeneration and collagen accumulation and fifibrosis at liver were observed. Compared with the model control, DBD group showed slighter hepatic fatty degeneration and collagen deposition, and had lower levels of ALT, AST and TBIL activities, lower contents of MDA, TG and Hyp, but higher SOD level and Alb content (P<0.05), and DBD also down-regulated MMP-2 mRNA expression and decreased MMP-2/9 activities in the fifibrotic livers (P<0.01).

CONCLUSIONThe action of DBD against liver fibrosis is related to prevent lipid peroxidation and inhibit MMP-2/9 activities in the fibrotic livers.

Animals ; Carbon Tetrachloride ; toxicity ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hepatic Stellate Cells ; drug effects ; Lipid Peroxidation ; drug effects ; Liver ; drug effects ; pathology ; physiopathology ; Liver Cirrhosis, Experimental ; drug therapy ; metabolism ; pathology ; Male ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Matrix Metalloproteinase Inhibitors ; Rats ; Rats, Wistar

OBJECTIVETo investigate the effects of Fuzheng Huayu recipe and Huangqi tang on DMN-induced experimental liver cirrhosis in rats and explore the therapeutic characteristics of Buxu herbals on liver cirrhosis.

METHODLiver cirrhosis in rats was induced by intraperitoneally injection of DMN for 4 weeks. Cirrhotic rats were randomly divided into 4 groups: model group, and Fuzheng Huayu recipe group, Huangqi tang group, Fuzheng Huayu recipe combined with Huangqi Tang group. The rats in treatment groups were orally administered with Fuzheng Huayu recipe, Huangqi tang, Fuzheng Huayu recipe combined with Huangqi tang (1:1), respectively. Normal and model control rats were given the equivalent normal saline. The body weight, liver weight and spleen weight were observed when rats were sacrificed. Liver histology was examined by HE staining and Sirius red staining. The liver function parameters including ALT, T. Bil and Alb were determined. The SOD activity and MDA content in liver tissues were also measured. Hepatic hydroxyproline (Hyp) content was determined by Jamall's method. The expression of alpha-SMA was determined by both immunohistochemistry staining and western blot method.

RESULTCompared with normal rats, the serum ALT and T. Bil levels in model rats increased obviously, by contrast, the serum Alb level decreased. There was a significant decline of SOD activity in model rat liver tissue, while the content of MDA and Hyp increased remarkably. A severe deterioration of liver architecture, infiltration of inflammatory cells and deposition of collagen were observed in model rat liver tissue. The expression of alpha-SMA also increased significantly. Compared with model rats, the liver function, lipid peroxidation parameters, Hyp content and liver histology were all improved in the 3 treatment groups. The combined group is better than any single-use group in decreasing collagen deposition and expression of alpha-SMA.

CONCLUSIONFuzheng Huayu recipe, Huangqi tang, Fuzheng Huayu recipe combined with Huangqi tang can attenuate liver fibrosis in DMN induced rats. Fuzheng Huayu recipe combined with Huangqi tang is better than that using alone in decreasing collagen deposition. The mechanism is partially due to the better effect of Fuzheng Huayu recipe combined with Huangqi tang on inhibiting activated HSC.

Animals ; Body Weight ; drug effects ; Dimethylnitrosamine ; adverse effects ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Liver ; drug effects ; enzymology ; Liver Cirrhosis, Experimental ; chemically induced ; drug therapy ; enzymology ; physiopathology ; Organ Size ; drug effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

Magnesium lithospermate B (MLB) is one of the major active components of Salvia miltiorrhizae. The anti-oxidative effects of Salvia miltiorrhizae have been previously reported. The aim of this study was to investigate the effect of purified MLB on hepatic fibrosis in rats and on the fibrogenic responses in hepatic stellate cells (HSCs). Hepatic fibrosis was induced in rats by intraperitoneal thioacetamide (TAA) injections over a period of 8 or 12 weeks. MLB was orally administered daily by gavage tube. Serum AST and ALT levels in TAA + MLB group were significantly lower than those in TAA only group at week 8. Hepatic fibrosis was significantly attenuated in TAA + MLB group than in TAA only group at week 8 or 12. Activation of HSCs was also decreased in TAA + MLB group as compared to TAA only group. Hepatic mRNA expression of alpha-smooth muscle actin (alpha-SMA), TGF-beta1, and collagen alpha1(I) was significantly decreased in TAA + MLB group as compared to TAA only group. Incubation with HSCs and MLB (> or =100 microM) for up to 48 h showed no cytotoxicity. MLB suppressed PDGF-induced HSC proliferation. MLB inhibited NF-kappaB transcriptional activation and monocyte chemotactic protein 1 (MCP-1) production in HSCs. MLB strongly suppressed H2O2-induced reactive oxygen species (ROS) generation in HSCs, and MLB inhibited type I collagen secretion in HSCs. We concluded that MLB has potent antifibrotic effect in TAA-treated cirrhotic rats, and inhibits fibrogenic responses in HSCs. These data suggest that MLB has potential as a novel therapy for hepatic fibrosis.
Actins/genetics/metabolism ; Animals ; Antioxidants/*administration & dosage ; Cell Proliferation/drug effects ; Collagen Type I/genetics/metabolism ; Drugs, Chinese Herbal/*administration & dosage ; Fibrosis/prevention & control ; Hepatic Stellate Cells/*drug effects/metabolism/pathology ; Liver/*drug effects/metabolism/pathology ; Liver Cirrhosis, Experimental/chemically induced/*drug therapy/physiopathology ; Male ; NF-kappa B/metabolism ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species/metabolism ; Salvia miltiorrhiza/immunology ; Thioacetamide/administration & dosage ; Transcriptional Activation/drug effects