Korean J Pathol.  2009 Dec;43(6):503-508. 10.4132/KoreanJPathol.2009.43.6.503.

Transcriptional Regulation of Hepatic Stellate Cell Activation by siRNA for TGF-beta1

• Oh HK
• Kim KH
• Keum YS
• Cho CH
• Park JB
• Park KK

• KMID: 1472634
• DOI: http://doi.org/10.4132/KoreanJPathol.2009.43.6.503

Abstract

BACKGROUND
The cytokine-induced activation of hepatic stellate cells (HSC) plays a major role in liver fibrosis. Quiescent HSCs undergo phenotypic transformation called "transdifferentiation" in response to viral, chemical or immune insults to the liver. The cytokine TGF-beta1 plays a key role in progressive liver fibrosis. Since small interfering RNA (siRNA) is a powerful tool for silencing gene expression post-transcriptionally, the present study aimed to determine whether synthetic TGF-beta1 siRNA down-regulates the expression of the TGF-beta1 gene in immortalized and activated rat HSCs (HSC-T6s). The study examined whether synthetic TGF-beta1 siRNA prevents rat HSCs activation and extracellular matrix (ECM) production. METHODS: TGF-beta1 siRNA or a control (pU6) siRNA was added to HSC-T6 culture media. We then performed RT-PCR and western blot analyses for TGF-beta1 and ECM components (fibronectin, type-I collagen, and TIMP-1). RESULTS: TGF-beta1 siRNA significantly down-regulated expression of TGF-beta1 mRNA and protein and attenuated mRNA and protein expressions of type-I collagen, fibronectin, and TIMP-1, as compared to the control. CONCLUSIONS: TGF-beta1 siRNA can effectively down-regulate the expression of TGF-beta1 in rat HSC, resulting in significant inhibition of HSC activation and of ECM production. These data indicate that synthetic TGF-beta1 siRNA can be a useful treatment modality to prevent liver fibrosis.

Keyword

Transforming growth factor-beta; Small interfering RNA; Hepatic stellate cell; Liver fibrosis

MeSH Terms

Animals
Blotting, Western
Collagen
Culture Media
Extracellular Matrix
Fibronectins
Gene Expression
Hepatic Stellate Cells
Liver
Liver Cirrhosis
Rats
RNA, Messenger
RNA, Small Interfering
Tissue Inhibitor of Metalloproteinase-1
Transforming Growth Factor beta1
Collagen
Culture Media
Fibronectins
RNA, Messenger
RNA, Small Interfering
Tissue Inhibitor of Metalloproteinase-1
Transforming Growth Factor beta1