cytochalasin-b and Cholestasis

The mechanism of cytochalasin B-induced intrahepatic cholestasis was examined using electron cytochemical techniques. Since previous studies suggested that the earliest lesions were in hepatic canaliculi, markers were used for three canalicular membrane components, namely ruthenium red for the glycoprotein-rich surface coat, the Mg2+-ATPase reaction as an example of a membrane-bound protein, and uranyl acetate en bloc and ruthenium red staining for the canalicular membrane-associated microfilaments. In rat liver infused in vivo with cytochalasin B, reduction in bile flow correlated with bile canalicular dilation, loss of the ruthenium red-positive surface coat from the canalicular membrane, and loss of demonstrable Mg2+-ATPase activity. In addition, structural alterations in microfilaments with widening of the ectoplasmic zone were noted. In isolated liver cells in vitro, identical changes were found. Bile canaliculi isolated from the in vivo cytochalasin B-infused rat liver lacked their normal investment of microfilaments. Detachment of the filaments from the bile canalicular membrane may be involved in the mechanism of cytochalasin B-induced cholestasis.

Topics: Adenosine Triphosphatases; Animals; Bile Ducts; Binding Sites; Cholestasis; Cytochalasin B; Cytoskeleton; Glycoproteins; Intercellular Junctions; Liver; Membrane Proteins; Microvilli; Rats; Ruthenium Red; Time Factors

The effects of cytochalasin B on bile canalicular structure and function were examined. Three experimental models were used, cultured hepatocytes, isolated perfused liver, and in vivo infused liver. The techniques used were light and electron microscopy and, in selected instances, scanning electron microscopy, electron "stains" for microfilaments, and measurements of bile flow. Microfilament disruption and dilation of bile canaliculi were consistently found and closely paralleled a reduction in bile flow in both in vitro and in vivo infused animals. It is proposed that under normal circumstances, the microfilaments maintain the canaliculi in a contracted or partly contracted state. Hence, the microfilamentous network would provide tone to the canalicular system which would tend to reduce stagnation and facilitate the flow of bile. Removal of normal microfilament contractile function would be expected to produce canalicular ectasia and reduction of bile flow, as was observed. Microfilament dysfunction may therefore be a possible cause of intrahepatic cholestasis. Crucial to this hypothesis are the presence of actin-containing microfilaments in the pericanalicular web, and an action of cytochalasin B on their contractility. Evidence pertaining to these requirements is presented and discused.

Topics: Animals; Bile; Bile Ducts, Intrahepatic; Cells, Cultured; Cholestasis; Cytochalasin B; Cytoplasm; Dilatation; Dimethyl Sulfoxide; Horses; Liver; Microscopy, Electron; Microscopy, Electron, Scanning; Perfusion; Rats