5-Hydroxymethylfurfural （5-HMF）, a water-soluble compound extracted from wine-processed Fructus corni, is a novel hepatic protectant for treating acute liver injury. The present study was designed to investigate the...5-Hydroxymethylfurfural （5-HMF）, a water-soluble compound extracted from wine-processed Fructus corni, is a novel hepatic protectant for treating acute liver injury. The present study was designed to investigate the protective effect of 5-HMF in human L02 hepatocytes injured by D-galactosamine （GAIN） and tumor necrosis factor-α （TNF-α） in vitro and to explore the underlying mechanisms of action. Our results showed that 5-HMF caused significant increase in the viability of L02 cells injured by GalN/TNF-α, in accordance with a dose-dependent decrease in apoptotic cell death confirmed by morphological and flow cytometric analyses. Based on immunofluorescence and Western blot assays, we found that GalN/TNF-α induced ER stress in the cells, as indicated by the disturbance of intracellular Ca^2＋ concentration, the activation of protein kinase RNA （PKR）-like ER kinase （PERK）, phosphorylation of eukaryotic initiation factor 2 alpha （eIF2α）, and expression of ATF4 and CHOP proteins, which was reversed by 5-HMF pre-treatment in a dose-dependent manner. The anti-apoptotic effect of 5-HMF was further evidenced by balancing the expression of Bcl-2 family members. In addition, the knockdown of PERK suppressed the expression of phospho-PERK, phospho-eIF2α, ATF4, and CHOP, resulting in a significant decrease in cell apoptosis after the treatment with GalN/TNF-α. 5-HMF could enhance the effects of PERK knockdown, protecting the cells against the GalN/TNF-α insult. In conclusion, these findings demonstrate that 5-HMF can effectively protect GalN/TNF-α-injured L02 hepatocytes against ER stress-induced apoptosis through the regulation of the PERK- eIF2α signaling pathway, suggesting that it is a possible candidate for liver disease therapy.展开更多
基金supported by grants from Natural Science Foundation of Jiangsu Higher Education Institutions of China(No.13KJB360010)Jiangsu Collaborative Innovation Center of Traditional Chinese Medicine(TCM) Prevention and Treatment of Tumor(No.012092002002)+1 种基金China and Europe Taking Care of Healthcare Solutions,CHETCH(No.PIRSES-GA-2013-612589)Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)
文摘5-Hydroxymethylfurfural （5-HMF）, a water-soluble compound extracted from wine-processed Fructus corni, is a novel hepatic protectant for treating acute liver injury. The present study was designed to investigate the protective effect of 5-HMF in human L02 hepatocytes injured by D-galactosamine （GAIN） and tumor necrosis factor-α （TNF-α） in vitro and to explore the underlying mechanisms of action. Our results showed that 5-HMF caused significant increase in the viability of L02 cells injured by GalN/TNF-α, in accordance with a dose-dependent decrease in apoptotic cell death confirmed by morphological and flow cytometric analyses. Based on immunofluorescence and Western blot assays, we found that GalN/TNF-α induced ER stress in the cells, as indicated by the disturbance of intracellular Ca^2＋ concentration, the activation of protein kinase RNA （PKR）-like ER kinase （PERK）, phosphorylation of eukaryotic initiation factor 2 alpha （eIF2α）, and expression of ATF4 and CHOP proteins, which was reversed by 5-HMF pre-treatment in a dose-dependent manner. The anti-apoptotic effect of 5-HMF was further evidenced by balancing the expression of Bcl-2 family members. In addition, the knockdown of PERK suppressed the expression of phospho-PERK, phospho-eIF2α, ATF4, and CHOP, resulting in a significant decrease in cell apoptosis after the treatment with GalN/TNF-α. 5-HMF could enhance the effects of PERK knockdown, protecting the cells against the GalN/TNF-α insult. In conclusion, these findings demonstrate that 5-HMF can effectively protect GalN/TNF-α-injured L02 hepatocytes against ER stress-induced apoptosis through the regulation of the PERK- eIF2α signaling pathway, suggesting that it is a possible candidate for liver disease therapy.
