Mitochondrion-localized retinol dehydrogenase 13 (Rdh13) is a short-chain dehydrogenase/reductase involved in vitamin A metabolism in both humans and mice. We previously generated Rdh13 knockout mice and showed that Rdh13 deficiency causes severe acute retinal light damage. In this study, considering that Rdh13 is highly expressed in mouse liver, we further evaluated the potential effect of Rdh13 on liver injury induced by carbon tetrachloride (CCl). Although Rdh13 deficiency showed no significant effect on liver histology and physiological functions under regular culture, the Rdh13 mice displayed an attenuated response to CCl-induced liver injury. Their livers also exhibited less histological changes and contained lower levels of liver-related metabolism enzymes compared with the livers of wild-type (WT) mice. Furthermore, the Rdh13 mice had Rdh13 deficiency and thus their liver cells were protected from apoptosis, and the quantity of their proliferative cells became lower than that in WTafter CCl exposure. The ablation of Rdh13 gene decreased the expression levels of thyroid hormone-inducible nuclear protein 14 (Spot14) and cytochrome P450 (Cyp2e1) in the liver, especially after CCl treatment for 48 h. These data suggested that the alleviated liver damage induced by CCl in Rdh13 mice was caused by Cyp2e1 enzymes, which promoted reductive CCl metabolism by altering the status of thyroxine metabolism. This result further implicated Rdh13 as a potential drug target in preventing chemically induced liver injury.
Alcohol Oxidoreductases ; deficiency ; genetics ; Animals ; Carbon Tetrachloride Poisoning ; enzymology ; Chemical and Drug Induced Liver Injury ; enzymology ; pathology ; Cytochrome P-450 CYP2E1 ; metabolism ; Female ; Immunohistochemistry ; Liver ; drug effects ; enzymology ; pathology ; Male ; Mice ; Mice, 129 Strain ; Mice, Inbred C57BL ; Mice, Knockout ; Nuclear Proteins ; metabolism ; Transcription Factors ; metabolism

Brown algae are well known as a source of biologically active compounds, especially those having antioxidant activities, such as phlorotannins. In this study we examined the antioxidant activities of crude phlorotannins extracts (CPEs) obtained from Sargassum hemiphyllum (SH) and fractionated according to the molecular weights. When CPEs were administrated at a dose of 30 mg/kg to Kunming mice pre-treated with carbon tetrachloride (CCl4), the levels of oxidative stress indicators in the liver, kidney and brain were significantly reduced in vivo. All the components of various molecular weight fractions of CPEs exhibited greater scavenging capacities in clearing hydroxyl free radical and superoxide anion than the positive controls gallic acid, vitamin C and vitamin E. Particularly, the components greater than 30 kD obtained from ethyl acetate phase showed the highest antioxidant capacities. These results indicated that SH is a potential source for extracting phlorotannins, the algal antioxidant compounds.
Animals ; Antioxidants ; isolation & purification ; pharmacology ; Ascorbic Acid ; pharmacology ; Brain ; drug effects ; metabolism ; pathology ; Carbon Tetrachloride ; antagonists & inhibitors ; toxicity ; Carbon Tetrachloride Poisoning ; drug therapy ; metabolism ; pathology ; Chemical Fractionation ; methods ; Gallic Acid ; pharmacology ; Hydroxyl Radical ; antagonists & inhibitors ; metabolism ; Kidney ; drug effects ; metabolism ; pathology ; Liquid-Liquid Extraction ; methods ; Liver ; drug effects ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred Strains ; Oxidation-Reduction ; Oxidative Stress ; drug effects ; Phaeophyta ; chemistry ; Sargassum ; chemistry ; Superoxides ; antagonists & inhibitors ; metabolism ; Tannins ; isolation & purification ; pharmacology ; Vitamin E ; pharmacology

The aim of the present study was to determine the preventive effects of the polysaccharide of Larimichthys crocea swim bladder (PLCSB) on CCl4-induced hepatic damage in ICR mice. The in vitro preventive effects of PLCSB on CCl4-induced liver cytotoxic effect were evaluated in BRL 3A rat liver cells using the MTT assay. The serum levels of AST, ALT, and LDH in mice were determined using commercially available kits. The levels of IL-6, IL-12, TNF-α, and IFN-γ were determined using ELISA kits. The pathological analysis of hepatic tissues was performed with H and E staining, and the gene and protein expressions were determined by RT-PCR and Western blotting, respectively. PLCSB (20 μg·mL(-1)) could increase the growth of BRL 3A rat liver cells treated with CCl4. The serum levels of AST, ALT, and LDH were significantly decreased when the mice were treated with two doses of PLCSB, compared with the control mice (P < 0.05). PLCSB-treated groups also showed reduced levels of the serum pro-inflammatory cytokines IL-6, IL-12, TNF-α, and IFN-γ. PLCSB could decrease the liver weight, compared to the CCl4-treated control mice. The histopathology sections of liver tissues in the 100 mg·kg(-1) PLCSB group indicated that the animals were recovered well from CCl4 damage, but the 50 mg·kg(-1) PLCSB group showed necrosis to a more serious extent. The 100 mg·kg(-1) PLCSB group showed significantly decreased mRNA and protein expression levels of NF-κB, iNOS, and COX-2, and increased expression of IκB-α compared with the CCl4-treated control group. In conclusion, PLCSB prevented from CCl4-induced hepatic damage in vivo.
Animal Structures ; chemistry ; Animals ; Biological Products ; pharmacology ; therapeutic use ; Carbon Tetrachloride ; Carbon Tetrachloride Poisoning ; drug therapy ; metabolism ; pathology ; Chemical and Drug Induced Liver Injury ; metabolism ; pathology ; prevention & control ; Cyclooxygenase 2 ; metabolism ; Cytokines ; blood ; I-kappa B Proteins ; metabolism ; Inflammation Mediators ; blood ; Liver ; drug effects ; metabolism ; pathology ; Male ; Mice, Inbred ICR ; NF-KappaB Inhibitor alpha ; NF-kappa B ; metabolism ; Necrosis ; Nitric Oxide Synthase Type II ; metabolism ; Perciformes ; Polysaccharides ; pharmacology ; therapeutic use ; RNA, Messenger ; metabolism

OBJECTIVETo investigate the effects of ancient Chinese medical formula Xiayuxue Decoction ([symbols; see text], XYXD) on activation of hepatic stellate cells (HSCs) and defenestration of sinusoidal endothelial cells (SECs) in CCl4-induced fibrotic liver of mice.

METHODSHigh performance liquid chromatography was used to identify the main components of XYXD and control the quality of extraction. C57BL/6 mice were induced liver fibrosis by CCl4 exposure and administered with XYXD for 6 weeks simultaneously. Liver tissue was investigated by hematoxylin-eosin and Sirius-red staining. Sinusoidal fenestrations were observed by scanning electronic microscopy and fluorescent immunohistochemistry of PECAM-1 (CD31). Whole liver lysates were detected of α-smooth muscle actin (α-SMA) and type-I collagen by Western blot. Primary rat HSCs-T6 cells were analyzed by detecting α-SMA, F-actin, DNA fragmentation through confocal microscopy, Western blot, terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) assay and cellomics arrayscan, respectively.

RESULTSAmygdalin and emodin in XYXD were identified. XYXD (993 mg/kg) inhibited Sirius red positive area up to 70.1% (P<0.01), as well as protein levels of α-SMA and type-I collagen by 42.0% and 18.5% (P<0.05) respectively. In vitro, XYXD (12.5 μg/mL, 50 μg/mL) suppressed the activation of HSCs and reversed the myofibroblastic HSCs into quiescent, demonstrated as inhibition of fluorescent F-actin by 32.3% and 46.6% (P<0.05). Besides, XYXD induced the apoptosis of HSC-T6 cells by 20.0% (P<0.05) and 49.5% (P<0.01), evidenced by enhanced TUNEL positivity. Moreover, ultrastructural observation suggested XYXD inhibited defenestration of SECs, which was confirmed by 31.1% reduction of protein level of CD31 (P<0.05).

CONCLUSIONSXYXD inhibited both HSCs activation and SECs defenestration which accompany chronic liver injuries. These data may help to understand the underlying mechanisms of XYXD for prevetion of chronic liver diseases.

Actins ; metabolism ; Animals ; Carbon Tetrachloride Poisoning ; drug therapy ; Collagen Type I ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Endothelium ; drug effects ; pathology ; Hepatic Stellate Cells ; drug effects ; pathology ; ultrastructure ; Liver Cirrhosis ; chemically induced ; drug therapy ; pathology ; Male ; Mice, Inbred C57BL ; Microscopy, Electron, Scanning ; Myofibroblasts ; drug effects ; pathology ; ultrastructure ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; Primary Cell Culture ; Rats, Sprague-Dawley

To evaluate the value of two histological quantitative methods of hepatic fibrosis: semiquantative scoring system (SSS) and image analysis by computer. The prophylactic and therapeutic effect of Ganzhifu on hepatic fibrosis induced by CCl4 were studied on a total 73 of specimens from liver tissue of rats. All specimen were analyzed quantatively by two methods of SSS marks and image analysis respectively. Difference between groups was compared and hydroxyproline (Hyp) content of each liver tissue was examined. Correlation analysis was done between SSS marks, image analysis and Hyp content. Both prophylactic and therapeutic study showed the same information. Results of SSS marks, image analysis and Hyp content were coincidence. It suggest that both SSS marks and image analysis were interrelated well with Hyp content (P < 0.01). The result suggests that both SSS marks of hepatic fibrosis and image analysis by computer can be taken as reliable histological quantitative method of hepatic fibrosis.
Animals ; Carbon Tetrachloride ; Carbon Tetrachloride Poisoning ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Hydroxyproline ; analysis ; Image Processing, Computer-Assisted ; Liver ; chemistry ; pathology ; Liver Cirrhosis, Experimental ; chemically induced ; drug therapy ; pathology ; Male ; Phytotherapy ; Rats ; Rats, Wistar

OBJECTIVETo investigate the role of perfusion-weighted magnetic resonance imaging (MRI) in evaluation of cirrhotic liver.

METHODSWith a 4F catheter, 1% diluted carbon tetrachloride (1 ml/kg) was selectively injected into right or left hepatic artery of 12 dogs fortnightly. The half liver into which carbon tetrachloride was injected was called as study side (SS), while the other half liver without carbon tetrachloride injection was called as study control side (SCS). Conventional and perfusion-weighted MRI were performed in every 4 weeks. Via a 4F catheter, 5ml gadolinium diethylentriamine pentaaceti acid (Gd-DTPA) dilution was injected into superior mesenteric artery at the 5th scan. The signal intensity-time curves of SS, SCS, and portal vein were completed in MR workstation. The maximal relative signal increase (MRSI), peak time (tp), and slope of the curves were measured.

RESULTSOn conventional MR images, no abnormalities of externality and signal intensity were observed in both SS and SCS of liver at each stage. The mean tp, MRSI, and slope of intensity-time curves in normal liver were 10.56 seconds, 1.01, and 10.23 arbitrary unit (au)/s, respectively. Three parameters of curves didn't show obvious change in SCS of liver at every stage. Abnormal perfusion curves occurred in SS of liver at the 12th week after the 1st injection. The abnormality of perfusion curve in SS was more and more serious as the times of injection increased. The mean tp, MRSI, and slope intensity-time curves in SS of liver were 19.45 seconds, 0.43, and 3.60 au/s respectively at the 24th week.

CONCLUSIONPerfusion-weighted imaging can potentially provide information about portal perfusion of hepatic parenchyma, and to some degree, reflect the severity of cirrhosis.

Animals ; Carbon Tetrachloride Poisoning ; Dogs ; Gadolinium DTPA ; Image Enhancement ; instrumentation ; Liver ; pathology ; ultrastructure ; Liver Circulation ; physiology ; Liver Cirrhosis, Experimental ; diagnosis ; physiopathology ; Magnetic Resonance Imaging ; instrumentation ; methods

OBJECTIVETo study the changes of advanced glycation end products (AGEs) in different phases of a rat liver fibrosis model induced by CCl4, and the interventional effect of aminoguanidin (AG).

METHODSFifty-four SD rats were divided into three groups: a control group, a CCl4 model group and an intervention group. Their blood serum AGEs and hyaluronic acid (HA) and AGEs in their liver homogenates were measured. These measurements were correlatively assessed to the degrees of liver fibrosis at different phases of the rat model before and after the intervention with aminoguanidin.

RESULTSThe content of AGEs in their blood sera and liver homogenates, and the level of blood serum HA, and the score of liver fibrosis degree at week 12 in our rat liver fibrosis mode groups were significantly higher than those in the control group (P < 0.01). In the intervention group with aminoguanidin, these figures were lower than those in the liver fibrosis model group (P < 0.05). The content of AGEs in their blood sera and liver homogenates had a linear correlation with the level of HA in their blood sera.

CONCLUSIONThe contents of AGEs in their blood sera and liver homogenates were increased in the late phase of our rat liver fibrosis model. To some extent, the level of AGEs may reflect the fibrosis degree of the rat livers. Aminoguanidin has an interventional effect in our CCl4 induced rat liver fibrosis model.

Animals ; Carbon Tetrachloride ; Carbon Tetrachloride Poisoning ; Glycation End Products, Advanced ; metabolism ; Guanidines ; therapeutic use ; Liver ; metabolism ; pathology ; Liver Cirrhosis, Experimental ; chemically induced ; drug therapy ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

Animals ; Carbon Tetrachloride Poisoning ; Diabetes Mellitus, Experimental ; complications ; Liver Cirrhosis, Experimental ; chemically induced ; complications ; pathology ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVESTo study the effectiveness of the lidocaine test in evaluating the liver reserve of rats with experimental liver injury in different phases.

METHODS40 healthy male Wistar rats were divided randomly into experimental and control groups. Rats of the experimental group received subcutaneous CCl4 in oil injection, and rats of the control group received saline injections. Monoethylglycinexylidide (MEGX) test, common hepatic function tests and histological examination of the livers were performed on all the rats.

RESULTSWith the development of the severity in liver injury, the concentrations of the serum MEGX in lidocaine test decreased gradually, which were consistent with liver histological changes. However, the results from the common liver function tests were all abnormal in the experimental group and were not consistent with the liver histological changes.

CONCLUSIONThe results obtained from the MEGX test are more agreeable to liver histological changes than those from common liver function tests. The results from the MEGX test can represent liver histological changes concisely.

Animals ; Carbon Tetrachloride ; Carbon Tetrachloride Poisoning ; Lidocaine ; analogs & derivatives ; Liver ; pathology ; physiopathology ; Liver Cirrhosis, Experimental ; chemically induced ; pathology ; physiopathology ; Liver Function Tests ; Male ; Random Allocation ; Rats ; Rats, Wistar

OBJECTIVETo observe the prophylactic effect of curcumin on hepatic fibrosis and the number, location, apoptosis of activated hepatic stellate cells (HSCs) in the livers and to discuss the relationship between the prophylactic effects and activated HSC.

METHODSA rat model of hepatic fibrosis was established by intraperitoneal injection of carbon tetrachloride. Curcumin doses of 5 mg, 10 mg, 20 mg per 100 gram per 100g of body weight were given to three groups of the model rats. No curcumin was given to one group of the model rats and it served as the control. After eight weeks, all rats were sacrificed and their left liver lobes were examined histopathologically with H.E and Masson stainings. Grades of hepatic fibrosis were evaluated according to the SSS system. Activated HSC was detected by the alpha-SMA immunohistochemistry staining. HSC apoptosis was detected by double-stainings of terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labeling (TUNEL) and desmin immunohistochemistry staining.

RESULTSDegrees (SSS system scores) of hepatic fibrosis in the curcumin groups were all less severe in comparison with those of the control group. Activated HSCs in the livers of the rats of the control group increased significantly compared with that of the treatment groups, and also fewer apoptotic HSCs were detected in the control group. On the contrary, fewer activated HSCs and more apoptotic HSCs were detected in the curcumin groups compared with those of the control group. The degrees of the effects were curcumin dose-dependent.

CONCLUSIONSCurcumin can prevent hepatic fibrosis. It can inhibit activation and proliferation of HSCs and induce HSCs apoptosis, which may be the mechanism(s) contributing to the prophylactic effects of curcumin on hepatic fibrosis.

Animals ; Apoptosis ; drug effects ; Carbon Tetrachloride ; Carbon Tetrachloride Poisoning ; Curcumin ; therapeutic use ; Enzyme Inhibitors ; therapeutic use ; Hepatocytes ; pathology ; Liver Cirrhosis, Experimental ; pathology ; prevention & control ; Rats ; Rats, Sprague-Dawley