No abstract available.
Extracellular Matrix/*metabolism ; Fibrosis ; Humans ; Liver/blood supply/metabolism/*pathology ; Liver Cirrhosis/etiology/genetics/*metabolism

In acute injury, liver recovers completely without any scarring change or complication. However, large portion of liver is changed into fibrotic state by excessive production of extracellular matrix (ECM) under chronic injury. Excessive production of ECM results in hepatic fibrosis and repeated process of hepatic fibrosis progress into liver cirrhosis. Liver cirrhosis is an irreversible and terminal state of chronic liver disease and one of the major causes of death in Korea. To block the progression to liver cirrhosis, various studies in the field of virology and immunology have been proceeded. Recently, studies on the hepatic fibrogenesis have progressed with the development of molecular biology. Hepatic stellate cells (HSC) play a key role in the pathogenesis of hepatic fibrosis by producing ECM. The degree of hepatic fibrosis depends on the proliferation and activation of HSC and increased net production of collagen. Therefore, inhibition of HSC activation is one of the main ways to block the progression of hepatic fibrosis. Many kinds of factors such as oxidative stress, acetaldehyde, ascorbic acid, transforming growth factor-beta (TGF-beta) and carbon tetrachloride (CCl4) have been reported to activate HSC and stimulate collagen gene expression. Although there are no definite and effective antifibrogenic agents, possible candidates are antioxidants, interferon, retinoids such as beta-carotene, flavonoids, renin-angiotensin system inhibitors and peroxisome proliferator activated receptor-gamma (PPAR-gamma) agonists. We tried to evaluate the charateristics of HSC in order to develop agents that inhibit hepatic fibrogenesis.
Extracellular Matrix/*metabolism ; Fibrosis ; Humans ; Liver/blood supply/metabolism/*pathology ; Liver Cirrhosis/etiology/genetics/*metabolism

BACKGROUNDSomafostatin receptors (SSTRs) have been suggested to involve in mediating the effect of somatostatin on hepatic stellate cells (HSCs) in an activation-dependent way. We, therefore, try to investigate the relationship between expression of SSTRs and activation of rat HSCs.

METHODSHSCs were isolated from rats by in situ perfusion and single-step density gradient centrifugation. SSTR1-5 mRNA levels in the differentiated first passage HSCs were detected by means of a reverse transcription polymerase chain reaction. On the other hand, hepatic fibrosis was induced in adult male Sprague-Dawley rats by carbon tetrachloride intoxication, and the expression of SSTR1-5 in normal as well as fibrotic livers was measured by immunohistochemical staining.

RESULTSSSTR mRNA and SSTR could not be found in freshly isolated rat HSCs or normal rat liver. However, SSTR1-3 mRNA appeared as HSCs became wholly activated, and could also be identified on the membrane of activated HSCs in the perisinusoid space, fibrous septa, etc.

CONCLUSIONThe expression of SSTR1-3 in the rat HSC is closely related to its activation. This may reflect one of the main negative regulation mechanisms in the course of HSC activation.

Animals ; Liver ; cytology ; metabolism ; Liver Cirrhosis ; etiology ; Male ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Receptors, Somatostatin ; genetics

Erythropoietic protoporphyria (EPP) is an inherited disorder of the heme metabolic pathway that is characterized by accumulation of protoporphyrin in the blood, erythrocytes, and tissues, and cutaneous manifestations of photosensitivity, all resulting from abnormalities in ferrochelatase (FECH) activity due to mutations in the FECH gene. Protoporphyrin is excreted by the liver, and excess protoporphyrin leads to cholelithiasis with obstructive episodes and chronic liver disease, finally progressing to liver cirrhosis. Patients with end-stage EPP-associated liver disease require liver transplantation. We describe here a 31-year-old male patient with EPP who experienced acute-on-chronic liver failure and underwent deceased-donor liver transplantation. Surgical and postoperative care included specific shielding from exposure to ultraviolet radiation to prevent photosensitivity-associated adverse effects. The patient recovered uneventfully and was doing well 24 months after transplantation. Future prevention and treatment of liver disease are discussed in detail.
Acute Disease ; Adult ; End Stage Liver Disease/etiology/pathology/*therapy ; Ferrochelatase/genetics/metabolism ; Humans ; Liver Cirrhosis/diagnosis ; *Liver Transplantation ; Male ; Mutation ; Protoporphyria, Erythropoietic/complications/*diagnosis/pathology

OBJECTIVETo investigate the diagnostic value of histopathological changes in the liver of patients with neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD).

METHODSLiver specimens from 10 cases of NICCD were evaluated by hematoxylin-eosin stain, histochemistry and immunohistochemistry (EnVision method). SLC25A13 mutation analysis was performed to correlate with histopathology.

RESULTSMost specimens showed varying degrees of fat deposition in hepatocytes, necrotic inflammation, cholestasis and fibrosis (so-called tetralogy). The combination of the above four histological changes was highly characteristic for NICCD. With the progression of the disease, hepatic fibrosis deteriorated and ultimately led to cirrhosis.

CONCLUSIONSNICCD should be suspected in the presence of cholestasis during infancy. A liver biopsy must be performed to rule out other liver diseases. The tetralogy of the hepatic histopathological changes has a highly diagnostic value for NICCD, which is also practical for accurately assessing the degree of inflammation and fibrosis, and similarly the progression of hepatic cirrhosis.

Biopsy ; Calcium-Binding Proteins ; deficiency ; genetics ; metabolism ; Cholestasis, Intrahepatic ; etiology ; genetics ; pathology ; Disease Progression ; Female ; Hepatocytes ; pathology ; Humans ; Infant ; Liver ; pathology ; Liver Cirrhosis ; pathology ; Male ; Mitochondrial Membrane Transport Proteins ; genetics ; Mutation ; Organic Anion Transporters ; deficiency ; genetics ; metabolism

ATP Binding Cassette Transporter, Sub-Family G, Member 2 ; ATP-Binding Cassette Transporters ; genetics ; metabolism ; Adult ; Aged ; Biomarkers, Tumor ; metabolism ; Carcinoma, Hepatocellular ; etiology ; metabolism ; pathology ; Drug Resistance, Multiple ; Female ; Humans ; Immunohistochemistry ; Liver ; metabolism ; pathology ; Liver Cirrhosis ; complications ; metabolism ; pathology ; Liver Neoplasms ; etiology ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Proteins ; genetics ; metabolism ; Neoplasm Staging ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo study the role of S6K1 in the induction of SREBP1c in mouse hepatic cell by high glucose stimulation.

METHODSS6K1 shRNA recombinant adenovirus (S6K1Ax) was injected into tail vein of db/db mice and then hepatic triglycerol content was analyzed. Liver specimen were stained with HE. After transfection with S6K1Ax or pU6Ax, mouse hepatic AML12 cells were treated with high glucose, insulin or glucose and insulin, the expression of mSREBP1c was detected by RT-PCR. S6K1 protein was detected by Western blot.

RESULTSHepatic S6K1 protein in db/db mice was inhibited a week after S6K1Ax injection. Compared with the control group, hepatic triglycerol content of S6K1Ax group was decreased (0.65+/-0.02) mmol/L vs (0.56+/-0.01) mmol/L (t = 4.312, P less than 0.01), hepatocyte fat droplet and vaculor generation were also decreased, fatty liver was improved. The mSREBP1c expression in S6K1Ax transfected cells was lower than that in the control cells (0.03+/-0.01 vs 0.06+/-0.01, t = 5.624, P less than 0.01). Compared with the basal state, SREBP1c expression of both groups was increased on the insulin stimulation, S6K1Ax group was 0.06+/-0.02 (t = 8.452, P less than 0.01) and control group was 0.08+/-0.02 (t = 3.591, P less than 0.05). There is no difference between control and S6K1Ax group by glucose addition (P more than 0.05).

CONCLUSIONS6K1 acts on fatty synthesis by regulating mSREBP1c expression.

Adenoviridae ; genetics ; Animals ; Cell Line ; Fatty Acid Synthases ; genetics ; metabolism ; Gene Expression Regulation ; Glucose ; administration & dosage ; Insulin ; administration & dosage ; Liver ; metabolism ; pathology ; Liver Cirrhosis ; etiology ; metabolism ; pathology ; Mice ; Mice, Inbred Strains ; RNA Interference ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Ribosomal Protein S6 Kinases, 90-kDa ; genetics ; metabolism ; Staining and Labeling ; Sterol Regulatory Element Binding Protein 1 ; genetics ; metabolism ; Transfection ; Triglycerides ; metabolism

OBJECTIVE: To determine whether there is an impaired Akt and eNOS activation in cirrhotic livers, and to investigate the feasibility of transferring adenovirus-mediated Akt gene to the liver for portal hypertension. METHODS: Recombinant adenovirus Ad-myr-HA-Akt and Ad-EGFP were produced by homologoas recombination in 293 cells . The Methods of compound factor, carbon tetrachloride (CCl4), corn flour, and cholesterol plus alcohol were used to construct the hepatic cirrhosis rat models. Ten normal rats were served as a normal control group, and 40 cirrhotic rats were divided into 4 groups randomly: an untreated group, an Ad-myr-HA-Akt treated group, an Ad-EGFP group, and a saline group. Ad-myr-HA-Akt, Ad-EGFP, and saline were transduced into the Ad-myr-HA-Akt treated group, Ad-EGFP group, and saline group via the tail vein respectively. Portal vein pressure, mean arterial pressure, and heart rate were measured in all rats. Protein abundance and phosphorylation status of Akt and eNOS were examined by Western blot. Spectrophotometry was used to measure the NO level. Frozen sections of the liver, heart, lung, kidney, brain, spleen, and testis were made to examine the expression of enhanced green fluorescent protein (EGFP) by fluorescence microscopy on Day 3 in the Ad-EGFP group. RESULTS: The concentration of recombinant adenovirus Ad-myr-HA-Akt after the purification was 5.5 x 10(11)vp/mL and that of Ad-EGFP was 6.0 x 10(11)vp/mL. Akt and eNOS phosphorylations in the liver of cirrhotic rats were obviously impaired. Adenoviral delivery of myr-Akt restored eNOS phosphorylation, increased the NO level and decreased the portal pressure after 3 days of adenoviral infection. In contrast, the livers infected with Ad-EGFP and saline were not changed. The EGFP expression was mainly found under the fluorescence microscopy on the frozen section of liver. Very little fluorescence was detected in the lung and kidney; and there was no detectable EGFP in other organs. CONCLUSION There is an impaired Akt and eNOS activation in the cirrhotic livers; myr-Akt gene therapy can restore the Akt activation and NO production in the cirrhotic liver, suggesting that this therapy may be helpful in treating portal hypertension.
Adenoviridae ; genetics ; metabolism ; Animals ; Carbon Tetrachloride ; Carbon Tetrachloride Poisoning ; Genetic Therapy ; Hypertension, Portal ; etiology ; therapy ; Liver Cirrhosis, Experimental ; complications ; metabolism ; therapy ; Male ; Nitric Oxide Synthase Type III ; metabolism ; Proto-Oncogene Proteins c-akt ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Angiotensin II ; pharmacology ; Animals ; Base Sequence ; Cell Division ; Collagen ; biosynthesis ; Liver ; cytology ; metabolism ; Liver Cirrhosis ; etiology ; Molecular Sequence Data ; Proline ; metabolism ; RNA, Messenger ; analysis ; Rats ; Receptor, Angiotensin, Type 1 ; genetics

BACKGROUNDIn clinical liver transplantation, whether the delay of hepatic arterial ischaemia increases biliary fibrosis or not is controversial. We designed a liver transplantation model to test this controversy and explore its mechanism.

METHODSTwelve dogs were divided into two groups randomly: hepatic arterial ischaemia (HAI) and control groups. In HAI group, hepatic artery was perfused 60 minutes after portal perfusion, but in control group, hepatic arterial perfusion was simultaneous with portal perfusion. The pathological changes of intrahepatic bile ducts were observed. Transforming growth factor beta 1 (TGF-beta1), expressed in epithelial cells of intrahepatic bile duct, was detected by immunohistochemical streptoadividin-biotin complex method. Expressions of Smad3, P-Smad3 and the transcriptional levels of alpha smooth muscle actin (alpha-SMA) mRNA in intrahepatic bile ducts were detected by Western blotting and RT-PCR respectively.

RESULTSCompared with the control group, more collagen deposition and leucocytic infiltration could be seen in biliary vessel walls. Significantly more buffy particles, which are the proteins of TGF-beta1, could be seen in biliary epithelial cells. P-Smad3 and alpha-SMA mRNA (as ratio to corresponding beta-actin) in intrahepatic bile ducts were 1.82 +/- 0.18 and 1.86 +/- 0.73 respectively in HAI group, significantly higher than those in control group (0.59 +/- 0.09 and 0.46 +/- 0.18, respectively).

CONCLUSIONSHepatic arterial ischaemia could increase the deposition of collagen fibres, trigger the transdifferentiation of myofibroblasts in intrahepatic bile duct and might result in biliary fibrosis by activating the TGF-beta1 signalling pathway.

Actins ; genetics ; Animals ; Blotting, Western ; Disease Models, Animal ; Dogs ; Hepatic Artery ; Immunohistochemistry ; Ischemia ; complications ; metabolism ; Liver Cirrhosis, Biliary ; etiology ; metabolism ; Liver Cirrhosis, Experimental ; Liver Transplantation ; adverse effects ; Male ; Random Allocation ; Reverse Transcriptase Polymerase Chain Reaction ; Smad3 Protein ; metabolism ; Transforming Growth Factor beta1 ; metabolism