Alpha B-crystallin expression in human and rat hepatic stellate cells

• Published in: Journal of hepatology Vol. 35; no. 2; pp. 200 - 207
• Main Authors: Cassiman, David, Roskams, Tania, van Pelt, Jos, Libbrecht, Louis, Aertsen, Paula, Crabbé, Tina, Vankelecom, Hugo, Denef, Carl
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier B.V 01.08.2001; Elsevier
• Subjects: Actins - metabolism; Alpha B-crystallin; Alpha-smooth muscle actin; Animals; Biological and medical sciences; Cells, Cultured; Chemical and Drug Induced Liver Injury - genetics; Chemical and Drug Induced Liver Injury - metabolism; Crystallins - genetics; Crystallins - metabolism; D-galactosamine; Desmin; Desmin - metabolism; Digestive system; Galactosamine - toxicity; Gene Expression; Glial fibrillary acidic protein; Hepatic stellate cell; Human; Humans; Immunohistochemistry; Investigative techniques, diagnostic techniques (general aspects); Liver - cytology; Liver - drug effects; Liver - metabolism; Male; Medical sciences; Nucleic Acid Hybridization; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques; Quantitative real-time reverse transcription polymerase chain reaction; Rat; Rats; Rats, Inbred WKY; Rats, Wistar; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - genetics; RNA, Messenger - metabolism; Suppression subtractive hybridization; Up-Regulation - drug effects; Human; Suppression subtractive hybridization; Alpha B-crystallin; Quantitative real-time reverse transcription polymerase chain reaction; D-galactosamine; Rat; Glial fibrillary acidic protein; Alpha-smooth muscle actin; Hepatic stellate cell; Desmin; Spider cell; Immunohistochemistry; Liver; Suppression; Rodentia; Cell biology; Hepatic disease; Hybridization; In vitro; Pathology; In vivo; Vertebrata; Mammalia; Crystallin; Subtraction; Animal; Alpha effect; Digestive diseases; Early
• ISSN: 0168-8278; 1600-0641
• DOI: 10.1016/S0168-8278(01)00122-2

Background/Aims : We searched for factors implicated in early hepatic stellate cell (HSC) activation in diseased liver, by means of suppression subtractive hybridization (SSH). Methods : SSH was performed between messenger RNA (mRNA) from normal rat HSC and mRNA from HSC, isolated from rats with acute D -galactosamine (Gal)-induced hepatitis. Results : One of the potentially upregulated factors which we found was alpha B-crystallin (ABCRYS), a small heat-shock protein and a chaperone known to protect the cell against protein degradation in conditions of cellular stress and known to associate with various types of intermediate filaments. Upregulation of ABCRYS mRNA in HSC, following Gal-intoxication (3.5-fold) as well as by culturing the HSC on plastic (20–30-fold), was confirmed by quantitative real-time reverse transcription polymerase chain reaction. The expression of ABCRYS protein in human and rat HSC was demonstrated by immunohistochemistry, in vitro and in vivo, in normal and diseased liver. Double-staining co-localized ABCRYS immunoreactivity with alpha-smooth muscle actin immunoreactivity in human liver and with desmin immunoreactivity in rat liver. In vivo upregulation of ABCRYS protein following Gal-intoxication was also shown, by comparison with desmin expression. Conclusions : Human and rat HSC express ABCRYS mRNA and protein. Both are rapidly upregulated following HSC activation.