Prevention of CCl4-induced liver cirrhosis by ribbon antisense to transforming growth factor-β1

Abstract

Abstract. Transforming growth factor-ß1 (TGF-ß1) is an important mediator of tissue fibrosis, including liver cirrhosis. Ribbon-type antisense oligonucleotide to TGF-ß1 (TGF-ß1 RiAS) was designed and combined with cationic peptide derived from the nuclear localization signal of human immunodeficiency virus-1 Tat protein for enhanced cellular uptake. When Hepa1c1c7 cells were transfected with TGF-ß1 RiAS, the level of TGF-ß1 mRNA was reduced by >70%. TGF-ß1 RiAS, mismatched RiAS, and normal saline were each injected into mice via the tail vein, beginning the week after intraperitoneal CCl 4 injection and continuing for 7 weeks, in order to determine whether TGF-ß1 RiAS prevents the fibrotic changes induced by the CCl 4 injection. After 8 weeks of the experiment, all of the mice treated with TGF-ß1 RiAS survived, compared to 50% of the control group and 65% of the mismatched RiAS-treated group. Upon examining the biochemical effects on the liver, TGF-ß1 mRNA levels were reduced significantly only in the TGF-ß1 RiAS-treated group. Immunohistochemical studies showed a reduced accumulation of collagen and α-smooth muscle actin. Our experimental results suggest that ribbon antisense to TGF-ß1, with efficient uptake, effectively blocks the expression of TGF-ß1 and prevents fibrosis of the liver.

p-CREB-1 promotes hepatic fibrosis through the transactivation of transforming growth factor-β1 expression in rats

“…Furthermore, we examined the underlying regulatory mechanism as well as the role of upregulated p-CREB-1 in liver fibrosis. It is well known that TGF-β1 is a potent cytokine which promotes the activation of HSCs and liver fibrogenesis, and is highly expressed in activated HSCs and CCl 4 -induced hepatic fibrosis (16,20,21). Herein, we explored the association between p-CREB-1 and TGF-β1 in liver fibrosis.…”

“…We then examined the regulatory mechanism responsible for the effects of p-CREB-1 in liver fibrosis. It is well known that TGF-β1 may be induced by CCl 4 and plays a crucial role in rat liver fibrogenesis (16), thus it is of interest to explore the association between p-CREB-1 and TGF-β1. Herein, we used recombinant human TGF-β1 protein (10 ng/ml) to activate the HSCs, and found that the protein expression levels of p-CREB-1 were significantly increased in the TGF-β1-treated HSCs compared with the control (p<0.01) (Fig.…”

Extracellular signal-regulated kinases 1/2 suppression aggravates transforming growth factor-beta1 hepatotoxicity: a potential mechanism for liver injury in methionine-choline deficient-diet-fed mice

“…28,29 Also, TGF-beta1 ribbon-type antisense oligonucleotide effectively prevented liver fibrosis. 30 Elevated TGF-beta1 expression is one of the major features associated with non-alcoholic fatty liver disease in the MCD diet feeding model. In late disease stage, TGF-beta1 signaling contributes to stellate cell activation and (myo)fibroblast generation, the latter being responsible for ECM deposition and scar formation.…”

GFAP Promoter-Driven RNA Interference on TGF-β1 to Treat Liver Fibrosis

“…TGF-β1 knockout mice have shown reduced collagen accumulation in response to liver injury (16). Inhibition of TGF-β1 gene expression or its signaling has been shown to decrease fibrosis in experimental animal model (17,18). Therefore, in this study, we selected two potent siRNA sequences for converting into shRNA and then cloning into pSilencer 1.0 vector driven by a ubiquitous U6 promoter due to its relative ease of producing shRNA transcripts.…”