Transforming growth factor­ß1 reduces apoptosis via autophagy activation in hepatic stellate cells.

Autophagy is a metabolic process that is important in fibrogenesis, in which cellular components are degraded by lysosomal machinery. Transforming growth factor ß1 (TGF­ß1) is a potent fibrogenic cytokine involved in liver fibrosis; however, it remains elusive whether autophagy is regulated by TGF­ß1 in this process. In the present study, the function of TGF­ß1­mediated autophagy in the proliferation and apoptosis of hepatic stellate cells (HSCs) was investigated. A rat HSC cell line (HSC­T6) was incubated with or without TGF­ß1 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 TGF­ß1 and autophagy. HSC­T6 cell transient transfection was accomplished with a pLVX­AcGFP­N1­rLC3B­encoding plasmid. An MTS assay and flow cytometry were utilized to detect proliferation and apoptosis of HSC­T6 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 HSC­T6 cells treated with TGF­ß1 compared with cells subjected to serum deprivation. However, when HSC­T6 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. GFP­LC3 punctate markings were more prolific following TGF­ß1 treatment of HSC­T6 cells, indicating that TGF­ß1 may rescue HSC­T6 cells from serum deprivation and reduce apoptosis via autophagy induction. The present study elucidated the possible functions of TGF­ß1­mediated autophagy in the pathological process of liver fibrosis.

ß-Nerve growth factor attenuates hepatocyte injury induced by D-galactosamine in vitro via TrkA NGFR.

Nerve growth factor (NGF) regulates the proliferation, differentiation and survival of cells and is also involved in the wound healing and tissue remodeling processes. The biological effects of NGF are dependent upon receptor signal-mediating functions, which differ between cells. This study attempted to investigate the hepatoprotective effect and possible mechanism of ß-NGF on D-galactosamine (D-GalN)-injured human liver L-02 cell lines. We demonstrated that L-02 cells expressed the neurotrophin receptors tyrosine kinase-A nerve growth factor receptor (TrkA NGFR) and p75 pan-neurotrophin receptor (p75NTR). Recombinant human ß-NGF markedly reduced cell injury and promoted the proliferation of L-02 cells damaged by D-GalN. However, this proliferation effect was blocked by the anti-TrkA NGFR antibody. Lactate dehydrogenase (LDH) and malondialdehyde (MDA) were released at reduced levels in the L-02 cell culture supernatant pretreated with ß-NGF. Furthermore, the albumin (ALB) content in the cell medium and intracellular glutathione (GSH) levels were markedly augmented, and the permeability of the mitochondrial membrane of the L-02 cells was improved by ß-NGF. Our results suggested that exogenous ß-NGF protects L-02 cells from D-GalN-induced injury through the NGF/TrkA NGFR signaling pathway.