Short interfering RNA targetting NF-kappa B induces apoptosis of hepatic stellate cells and attenuates extracellular matrix production
| Autor: | Huiqing Jiang, Junji Ma, Donglai Cui, Sui Zhang, Jing Han |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
Small interfering RNA
Apoptosis Biology Matrix metalloproteinase Liver Cirrhosis Experimental Collagen Type I Transforming Growth Factor beta1 Extracellular matrix Hepatic Stellate Cells Animals Humans Gene Silencing RNA Small Interfering Cell Line Transformed Tissue Inhibitor of Metalloproteinase-1 Hepatology Caspase 3 Reverse Transcriptase Polymerase Chain Reaction NF-kappa B Transcription Factor RelA Gastroenterology Transfection Transforming growth factor beta Molecular biology Extracellular Matrix Cell culture Hepatic stellate cell biology.protein Cattle Apoptosis Regulatory Proteins |
| Zdroj: | Digestive and Liver Disease. 42:813-817 |
| ISSN: | 1590-8658 |
| DOI: | 10.1016/j.dld.2010.03.011 |
| Popis: | Background Pharmacological inhibition of the NF-κB activity enhances hepatic stellate cell apoptosis and reverses experimental fibrosis. However, there is no report on the effects of NF-κB knockdown on apoptosis and extracellular matrix secretion in hepatic stellate cells. The aim of the present study is to explore the effects of siRNA targetting NF-κB on the apoptosis and extracellular matrix production in hepatic stellate cells. Methods The immortalised hepatic stellate cell line HSC-T6 was transfected with siRNA; 72 h later, cells were stimulated by LPS for 1 h; these cells were collected for further use. Hepatic stellate cell apoptosis was determined by fluorescence activated cell sorter analysis, TUNEL assay and caspase-3 activity measurement. Matrix metalloproteinase 2 activity was evaluated with Gelatin zymography. The quantities of mRNA transcriptions of NF-κB p65, type I collagen, tissue inhibitor of metalloproteinases-1, α-smooth muscle actin and transforming growth factor beta 1 and anti-apoptotic protein A1 were evaluated with quantitative reverse transcriptase real-time polymerase chain reaction. Results siRNA targetting NF-κB p65 effectively abrogated the expression of NF-κB p65 in hepatic stellate cells; decreased anti-apoptotic protein Bcl-2 and the mRNA transcription of hepatic type I collagen, α-smooth muscle actin, transforming growth factor beta 1, A1 and tissue inhibitor of metalloproteinases-1; increased matrix metalloproteinase 2 activity and promoted hepatic stellate cell apoptosis. Conclusion NF-κB knockdown enhances hepatic stellate cell apoptosis and attenuates extracellular matrix production. |
| Databáze: | OpenAIRE |
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