Hepatic stellate cell (HSC) plays a pivotal role in liver fibrosis and isconsidered as one of the therapeutic targets for the treatment of hepatic fibrosis. Focal adhesion kinase (FAK) has been shown to play an important role in the HSC activation. The aim of the study is to explore the role of FAK in the proliferation and activation in culture-activated rat HSCs by using a specific antisense oligonucleotides targeting FAK (FAK-ASON). Rat HSCs were prepared from SD rats by in situ perfusion of pronase and collagenase and single-step density Nycodenze gradient. Culture-activated HSCs were transfected with the FAK-ASON (1 microM) by lipofectamine 2000 for 24, 48 or 72 hours. The proliferation of HSC was detected by MTT assay. The expression of the marker of HSC activation, alpha-smooth muscle actin (alpha-SMA), was assessed by reverse transcription- polymerase chain reaction (RT-PCR) and Western blot. The inhibition rates for HSC proliferation 24, 48 and 72 hours after transfection were 65.5% +/- 5.8%, 46.8% +/- 4.3% and 35.7% +/- 5.2% respectively. Transfection of FAK-ASON could significantly inhibit the proliferation of HSC. Meanwhile, treatment with the FAK-ASON could markedly decrease the mRNA and protein expression of alpha-SMA in rat HSC. The specific FAK-ASON may have an inhibitory effect on the proliferation and activation in rat HSC.
Actins ; biosynthesis ; genetics ; Animals ; Cell Proliferation ; Cells, Cultured ; Focal Adhesion Protein-Tyrosine Kinases ; metabolism ; Hepatocytes ; cytology ; enzymology ; Male ; Oligonucleotides, Antisense ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley ; Transfection