Cells with morphological and immunohistochemical features of hepatic stellate cells (Ito cells) form an extralittoral (extrasinusoidal) compartment in the cirrhotic rat liver.

Systematic studies on hepatic stellate cells and myofibroblasts have so far mainly focused on cells located in the perisinusoidal space of Disse, the so-called littoral compartment. Here, these cells play a key role for intralobular fibrogenesis and sinusoidal capillarization. However, advanced hepatic fibrosis and cirrhosis are characterized by portal tract fibrosis and septal fibrosis, thus involving cells outside the perisinusoidal space. To study the question as to whether hepatic stellate cells occur and are expanded in an extralittoral (extrasinusoidal) compartment in cirrhogenesis, we systematically analyzed the distribution and density of desminreactive stellate cells in a rat model of hepatic fibrosis. Fibrosis and remodeling of the liver were induced by bile duct ligation, and stellate cells were identified by single and double immunohistochemistry. We can show that desmin-reactive cells are reproducibly detectable in extralittoral compartments of the normal and fibrotic rat liver. Periductular extralittoral stellate cells are significantly more frequent in cirrhosis, indicating that extralittoral stellate cells expand in concert with proliferating ductules. The findings suggest that ductular proliferation thought to represent a pacemaker of hepatic remodeling is accompanied by a population of cells exhibiting the same phenotype as perisinusoidal stellate cells.

Hepatic stellate cells (Ito cells) but not collagen IV may partly be responsible for lower portal pressure after reversing secondary biliary cirrhosis in the rat.

BACKGROUND/AIMS: Chronic bile duct obstruction in the rat leads to biliary cirrhosis and portal hypertension. Biliary decompression with Rouxen-Y choledocho-jejunostomy (RY) reverses most but not all of these changes. The aim of the present study was to determine whether hepatic stellate cells, as a main source of extracellular matrix proteins, participate in this process. METHODS: Sprague-Dawley rats were allocated to one of three groups: Bile duct ligation (BDL) for 3 weeks, BDL followed by RY and sham-operated animals as controls (SHAM). At the end of the experimental period, portal pressure was measured, livers subjected to random sampling and hepatocytes, bile ducts/ductules, hepatic stellate cells and collagen IV determined stereologically. Hepatic stellate cells and collagen IV were characterized immunohistochemically with an antibody against desmin and collagen IV, respectively. RESULTS: Volume fraction of hepatocytes decreased from 65.6 +/- 5.3 in sham-operated animals to 27.9 +/- 8.8% in bile duct ligated animals (p < 0.05). In contrast, volume fraction of bile ducts/ductules increased from 0.4 +/- 0.2 in sham-operated animals to 25.3 +/- 8.6% in bile duct ligated ones; similarly, hepatic stellate cells increased from 0.4 +/- 0.2 in sham-operated animals to 2.6 +/- 0.9% in bile duct ligated ones (p < 0.01) and collagen IV from 10.0 +/- 2.3 in sham-operated animals to 24.5 +/- 8.0% (p < 0.01) in bile duct ligated animals. These changes were partially reversed by Roux-en-Y choledocho-jejunostomy; hepatocytes, bile ducts/ductules, hepatic stellate cells and collagen IV averaging 54.8 +/- 13.1, 6.1 +/- 6.8, 1.6 +/- 0.6 and 14.5 +/- 3.6%, respectively (p < 0.05 RY vs. BDL). Portal pressure in sham-operated animals, bile duct ligated animals and those with a Roux-en-Y choledocho-jejunostomy averaged 13.4 +/- 0.7, 20.1 +/- 2.7 and 16.9 +/- 1.6 cm H2O, respectively, and correlated significantly with the volume fraction of hepatic stellate cells (rS = 0.96; p < 0.001) and less with collagen IV (rS = 0.61; p < 0.007). However, by stepwise regression, collagen IV did not significantly add to the ability of the equation to predict portal pressure. CONCLUSIONS: These results lend further support to the notion that hepatic stellate cells are prominently involved in fibrogenesis and in the reversibility of these changes, but hepatic stellate cells do not completely revert to normal even 4 weeks after successful decompression. Furthermore, our data suggest that hepatic stellate cells may be related to maintenance of portal hypertension.

Involvement of structural and nonstructural polypeptides on rotavirus RNA synthesis.

Rotavirus are segmented double stranded RNA viruses with a double protein capsid around a central core. The virus replicates in the cell cytoplasm. After infection, eleven mRNAs are transcribed from the viral genome. To characterize further the infection cycle, viral polypeptide synthesis and RNA replication were studied using labelled precursors. The involvement of nonstructural polypeptides NS34 and NS35 was determined by the kinetics of the appearance of viral polypeptides in infected cells. Experiments in which cycloheximide was used showed that the synthesis of both polypeptides was required to begin RNA replication. The isolation of subviral particles at 8 hours postinfection indicates that there is a particle containing the nonstructural polypeptides and the structural polypeptides VP1, VP2, and VP6 that seem to be able to transcribe the viral genome to produce other RNA species. The results suggest that there is a core-like particle similar to one obtained in vitro which upon the addition of VP6 is able to transcribe the virus genome. This seems to indicate that core-like particles may alter their specificity for plus or minus RNA synthesis depending upon the polypeptides that interact with it. The interaction between VP6 and the viral core was analyzed by means of antibodies raised against the viral core and VP6. The results suggest that VP6 contains a specific binding site to the core complex and this interaction allows the synthesis of mRNA.