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Protective effect of Cornel iridoid glycoside (CIG) in GalN/TNF-α-injured L02 hepatocytes and its mechanism

Zequn Jiang, Yanxia Ma, Lihua Zhou, Haiying Jiang, Mingyan Wang, Xiuqin Zhan.

Abstract
Aim: To determine the action mode of Cornel iridoid glycoside (CIG) from Fructus corni on hepatoprotective activities, the effects of CIG on human hepatocyte cell line (L02) injured by D-galactosamine (GalN) and tumor necrosis factor-α (TNF-α) were examined.
Methods: The percentage of cell viability was evaluated by Cell Counting Kit-8 (CCK-8) assay. Apoptosis was detected by flow cytometric analysis in human L02 hepatocytes. The expression levels of activating transcription factor‑4 (ATF4), and C/EBP homologous protein (CHOP) were detected by western-blot analysis. In addition, the activity of caspase-3 was tested by enzyme-linked immunosorbent assay.
Results: The results showed that CIG caused significant increase in the viability of L02 cells injured by GalN/TNF-α, in accordance with a dose-dependent decrease of apoptotic cell death confirmed by flow cytometric analysis. Based on western blot and colorimetric assay, we found that GalN/TNF-α induced increased expression of ATF4, CHOP and activation of caspase-3, while CIG pre-treatment had a dose-dependent suppression on them in this cell model.
Conclusion: Overall, these findings demonstrate that CIG can effectively protect L02 hepatocytes against apoptosis induced by GalN/TNF-α, suggesting that it is a possible candidate target for liver disease therapy.

Key words: CIG, GalN/TNF-α, L02 hepatocyte, apoptosis


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