Anti-fibrotic Effects via Regulation of Transcription Factor Sp1 on Hepatic Stellate Cells

Cellular Physiology and Biochemistry

关键词: Transcription factor Sp1;    Decoy oligodeoxynucleotide;    Hepatic stellate cells;    Hepatic fibrosis;    Proliferation;   

DOI  :  10.1159/000337586

学科分类：分子生物学，细胞生物学和基因

来源: S Karger AG

【 摘 要 】

Background Hepatic stellate cells (HSCs), the central cells in hepatic fibrosis, are characterized by sustaining activation, a process that consists in increased proliferation and over-expression of fibrotic genes. Transcription factor Sp1 mediates the expression of a variety of fibrotic genes expression and thereby play an important role in fibrosis. In addition, previous reports have indicated that Sp1 binding activity is greatly increased in activated HSCs. Thus, our aim was to investigate the anti-proliferative and anti-fibrotic effects of the oligonuceotide decoy of the transcription factor Sp1, ODN, a potent inhibitor of Sp1-activated transcription. Methods We optimized Lipofectamin 2000 (LF2000)ODN DNA ratio for the transfection of hepatic stellate cells HSC-T6. Then we measure the effect of transfected ODN on HSC-T6 cells′ proliferation and fibrotic gene expression, and study the mechanism involved. Results At a DNA concentration of 1 µM and a ratio ODN DNALF2000 of 13, HSC-T6 cells have the maximal transfection efficiency with the lowest toxicity. Transfected ODN effectively blocks Sp1 binding to the promoter regions of cell cycle regulatory proteins cyclin D1, p27^KIP1 and fibrotic genes, including transforming growth factor (TGF)-β1, Platelet-derived growth factor (PDGF)-BB, α-SMA, α1 (I) collagen and tissue inhibitor of metalloproteinases-1 (TIMP-1). ODN inhibits HSC-T6 proliferation and fibrotic genes expression in vitro. Conclusion Sp1 is a key transcription factor that mediates proliferation and fibrotic gene synthesis of HSC-T6, inhibition of Sp1 with decoy ODN may be an effective approach to prevent the progression of hepatic fibrosis.

【 授权许可】

CC BY-NC-ND