Fetuin-A gene expression, synthesis and release in primary human hepatocytes cultured in a galactosylated membrane bioreactor.
This paper reports on human hepatocytes cultured in a galactosylated membrane bioreactor in order to explore the modulation of the effects of a pro-inflammatory cytokine, Interleukin-6 (IL-6) on the liver cells at molecular level. In particular the role of IL-6 on gene expression and production of a glycoprotein, fetuin-A produced by hepatocytes, was investigated by culturing hepatocytes in the membrane bioreactor, both in the absence and presence of IL-6 (300 pg/ml). IL-6 modulated the fetuin-A gene expression, synthesis and release by primary human hepatocytes cultured in the bioreactor. A 75% IL-6-induced reduction of fetuin-A concentration in the medium was associated with a 60% increase of C-reactive protein in the same samples. Real-time-PCR demonstrated an 8-fold IL-6-induced reduction of fetuin-A gene expression. These results demonstrate that the hepatocyte galactosylated membrane bioreactor is a valuable tool to study IL-6 effects and gave evidence, for the first time, that IL-6 down-regulates the gene expression and synthesis of fetuin-A by primary human hepatocytes. The human hepatocyte bioreactor behaves like the in vivo liver, reproducing the same hepatic acute-phase response that occurs during the inflammatory process.
Memoli
Bruno
B
Department of Nephrology, University Federico II of Naples, Napoli, Italy.
De Bartolo
Loredana
L
Favia
Pietro
P
Morelli
Sabrina
S
Lopez
Linda C
LC
Procino
Alfredo
A
Barbieri
Giuseppe
G
Curcio
Efrem
E
Giorno
Lidietta
L
Esposito
Pasquale
P
Cozzolino
Mario
M
Brancaccio
Diego
D
Andreucci
Vittorio E
VE
d'Agostino
Riccardo
R
Drioli
Enrico
E
D019149:Bioreactors; D001798:Blood Proteins / Q000235:genetics*; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D002478:Cells, Cultured; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005690:Galactose / Q000737:chemistry*; D015870:Gene Expression / Q000502:physiology; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism*; D006801:Humans; D015850:Interleukin-6 / Q000008:administration & dosage*; D019164:Liver, Artificial; D008567:Membranes, Artificial; D023822:Tissue Engineering / Q000295:instrumentation* / Q000379:methods; D060749:alpha-2-HS-Glycoprotein
C556354:AHSG protein, human; D001798:Blood Proteins; D015850:Interleukin-6; D008567:Membranes, Artificial; D060749:alpha-2-HS-Glycoprotein; D005690:Galactose