Journal
MOLECULAR MEDICINE REPORTS
Volume 10, Issue 3, Pages 1282-1288Publisher
SPANDIDOS PUBL LTD
DOI: 10.3892/mmr.2014.2383
Keywords
transforming growth factor-beta 1; hepatic stellate cell; autophagy; apoptosis
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Funding
- Wang Bao-en Foundation of Hepatic Fibrosis
- Chinese Foundation for Hepatitis Prevention and Control [CFHPC20131014]
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Autophagy is a metabolic process that is important in fibrogenesis, in which cellular components are degraded by lysosomal machinery. Transforming growth factor beta 1 (TGF-beta 1) is a potent fibrogenic cytokine involved in liver fibrosis; however, it remains elusive whether autophagy is regulated by TGF-beta 1 in this process. In the present study, the function of TGF-beta 1-mediated autophagy in the proliferation and apoptosis of hepatic stellate cells (HSCs). was investigated. A rat HSC cell line (HSC-T6) was incubated with or without TGF-beta 1 followed by bafilomycin A1, and microtubule-associated proteins 1A/1B light chain 3 (LC3) small interfering (si)RNA was used to inhibit autophagy in order to assess the association between TGF-beta 1 and autophagy. HSC-T6 cell transient transfection was accomplished with a pLVX-AcGFP-N1-rLC3B-encoding plasmid. An MTS assay and flow cytometry were utilized to detect proliferation and apoptosis of HSC-T6 cells. Quantitative polymerase chain reaction,immunofluorescence and western blot analysis were used to detect the presence of activation markers. Proliferation was increased and apoptosis was reduced in HSC-T6 cells treated with TGF-beta 1 compared with cells subjected to serum deprivation. However, when HSC-T6 cells were treated with bafilomycin A1 and LC3 siRNA, increased apoptosis and reduced proliferation were observed. In addition, protein and mRNA expression levels of the autophagy marker LC3 were significantly increased. GFP-LC3 punctate markings were more prolific following TGF-beta 1 treatment of HSC-T6 cells, indicating that TGF-beta 1 may rescue HSC-T6 cells from serum deprivation and reduce apoptosis via autophagy induction. The present study elucidated the possible functions of TGF-beta 1-mediated autophagy in the pathological process of liver fibrosis.
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