griseofulvin and Hepatitis--Alcoholic

Mallory bodies are a morphological key feature of severe alcoholic liver cell injury (alcoholic hepatitis) and the morphological expression of dysregulation and derangement of the intermediate filament cytoskeleton of the hepatocyte. Their pathogenesis is still unclear. Studies on Mallory body formation may not only help to elucidate the mechanisms of liver cell injury associated with alcoholic hepatitis, but may also contribute to our understanding of the regulation and function of the intermediate filament cytoskeleton.

Topics: Animals; Chemical and Drug Induced Liver Injury; Colchicine; Cytoskeleton; Electrophoresis, Polyacrylamide Gel; Endoplasmic Reticulum; Fluorescent Antibody Technique; Griseofulvin; Hepatitis, Alcoholic; Humans; Isoantibodies; Isoelectric Focusing; Keratins; Liver; Liver Cirrhosis, Alcoholic; Mice; Microscopy, Electron; Rats

Livers containing Mallory bodies (MBs, hyalin degenerative cytoplasmic inclusions) were examined using Heuser's and Van Harreveld's cryo-techniques. The tissues were collected from 1) a patient suffering from alcoholic hepatitis and 2) mice treated with griseofulvin (GF, an anti-mitotic drug). Normal mouse liver and isolated MBs from GF-treated mice were also analyzed by the same methods. Our results suggest that under the toxic influence of alcohol or GF on microtubular elements, MBs are generated by entanglement of elements of 10 nm filaments with microtubule elements. This in turn inhibits cellular transport processes. The reticular net of the ER-element which is usually observable in the normal tissue is changed into numerous small vesicles in the pathological and experimental tissues. The diameters of hepatocytes containing these vesicles were 1.5 to 2 times larger than control diameters. MBs have previously been described in thin sections as filamentous tangles. On replicas we found that they appear to be composed of pairs of filaments twisted in a roughly helical manner, each having a diameter less than 10 nm. The paired helical nature of the MB-filaments is reminiscent of other inclusion bodies, which are also composed of elements of 10 nm filaments, observable in various neurological diseases.

Topics: Actin Cytoskeleton; Adult; Alcohols; Animals; Female; Freeze Etching; Griseofulvin; Hepatitis, Alcoholic; Humans; Hyalin; Inclusion Bodies; Liver; Liver Cirrhosis, Experimental; Male; Mice; Mice, Inbred C3H; Microscopy, Electron; Microtubules; Neurons; Vacuoles

Mallory bodies (MBs) are characteristics morphologic features of alcoholic hepatitis and can be produced in mouse hepatocytes by chronic griseofulvin (GF) intoxication. The formation of MBs, which share some immunological, biochemical, and ultrastructural features with cytokeratin (CK) filaments of normal liver, is accompanied by derangement and even loss of the CK cytoskeleton of hepatocytes ("empty cells") as revealed by immunofluorescence microscopy. To clarify whether this diminution or lack of CK-related staining of MB-containing hepatocytes was due to loss of CK filaments or changes in antigenicity or accessibility of antigenic determinants immunohistochemical studies using a battery of monoclonal and polyclonal CK antibodies were performed. It could be shown that all these antibodies directed against different CK polypeptide components and antigenic determinants of CKs revealed a highly reduced or even undetectable cytoplasmic CK meshwork in most cells with fully developed large MBs. In the light of our present knowledge of the organization of CK intermediate filaments, these results indicate that the phenomenon of the "empty cells" reflects a diminution of CK meshwork rather than altered antigenic determinants.

Topics: Animals; Antibodies, Monoclonal; Bile Ducts, Intrahepatic; Desmin; Electrophoresis, Polyacrylamide Gel; Griseofulvin; Hepatitis, Alcoholic; Humans; Immunoblotting; Inclusion Bodies; Intermediate Filaments; Keratins; Liver; Male; Mice; Microscopy, Fluorescence; Vimentin

To identify Mallory body (MB) constituents, monoclonal antibodies to murine MBs induced by long-term griseofulvin (GF) feeding were produced. One of these, antibody MM 120-1, specifically reacted in immunofluorescence microscopy with MBs in all developmental stages but not with other cell structures of human and mouse liver and other organs. The MM 120-1 antigen was present in murine MBs induced by griseofulvin or 3,5-diethoxycarbonyl-1,4-dihydrocollidine feeding and also in human MBs in livers of patients with alcoholic hepatitis. In immunoblots, the MM 120-1 antigen was detectable in the high molecular weight fraction of MB preparations, most of which remained in the well and at the interphase between stacking and resolving gel. No reactivity with cytokeratin polypeptides of different conformational states (i.e., isolated cytoker atin components A and D, heterotetramers A2D2, reconstituted intermediate filaments) was found. It is concluded that the antibody MM 120-1 is a highly specific and sensitive marker for murine and human MBs recognizing a high molecular weight nonkeratin component. This component could play a central role in the pathogenesis of MBs.

Topics: Animals; Antibodies, Monoclonal; Cytoplasm; Dicarbethoxydihydrocollidine; Fluorescent Antibody Technique; Griseofulvin; Hepatitis, Alcoholic; Humans; Inclusion Bodies; Liver; Mice; Molecular Weight; Staining and Labeling

Topics: Animals; Antibodies; Cytoskeleton; Griseofulvin; Hepatitis, Alcoholic; Histocytochemistry; Humans; Immunodiffusion; Inclusion Bodies; Liver; Liver Cirrhosis; Mice; Mice, Inbred Strains; Rabbits

Antibodies raised against prekeratin intensely and specifically stain, in immunofluorescence microscopy, Mallory bodies ("alcoholic hyalin") present in livers of human alcoholics and griseofulyin-treated mice. The high sensitivity of this method allows the identification of small distinct cytoplasmic structures that are observed during early stages of Mallory body formation, especially frequent in the perinuclear cytoplasm, as well as during stages of Mallory body disintegration and disappearance, such as after withdrawal of the drug. In the latter situation, the prekeratin-containing small particles exhibit a characteristic pattern of arrangement in the hepatocyte periphery. Electron microscopy illustrates that such small bodies are heap-like aggregates of typical Mallory body filaments. Immunofluorescence studies with antibodies to isolated prekeratin polypeptides from bovine hoof or muzzle epidermis show that Mallory body filaments, in particular those in human liver, are immunologically more closely related to prekeratin of tonofilaments from living epidermal cells (stratum spinosum). The data indicate that Mallory bodies contain a pathologic form of prekeratin-like material. They also suggest that disorders of cytoskeletal structures of the intermediate-sized filament class are associated with specific diseases and can be visualized and characterized by immunofluorescence microscopy by using antibodies to constitutive proteins of such filaments.

Topics: Animals; Cytoplasmic Granules; Fluorescent Antibody Technique; Griseofulvin; Hepatitis, Alcoholic; Humans; Keratins; Liver Cirrhosis, Alcoholic; Liver Cirrhosis, Experimental; Mice; Protein Precursors

To investigate the possibility that Mallory bodies (MBs) are composed of intermediate filaments (IFs), the electrophoretic patterns of MB proteins from human liver were compared with the IF protein extracted from chicken gizzard smooth muscle and extracts of mouse liver which contained MBs. The human MB had a protein component which had a similar mobility as that of the IF protein extracted from smooth muscle. A similar protein band was found in the extracts of mouse liver which contained MBs. No protein band corresponding to actin protein was found in human MB protein. The IF-like protein was also present in control mouse liver extract. The estimated molecular weight of the IF protein was 54,000. The evidence suggests that MBs are, in part, composed of IF protein. The IF protein is a normal constituent of liver and smooth muscle.

Topics: Animals; Chickens; Electrophoresis, Polyacrylamide Gel; Endoplasmic Reticulum; Griseofulvin; Hepatitis, Alcoholic; Humans; Liver; Male; Mice; Mice, Inbred C3H; Molecular Weight; Muscle, Smooth; Proteins