Effect of silibinin on ethanol- or acetaldehyde-induced damge of mouse primary hepatocytes in vitro. (February 2021)

• Record Type: Journal Article
• Authors: Song, Xiao-Yu; Li, Rong-Hua; Liu, Wei-Wei; Hayashi, Toshihiko; Mizuno, Kazunori; Hattori, Shunji; Fujisaki, Hitomi; Ikejima, Takashi
• Is Part Of: Toxicology in vitro. Volume 70(2021)
• Publication Date: 2021-02
• Subjects: Silibinin -- Ethanol -- Acetaldehyde -- Hepatocytes -- Apoptosis -- Autophagy; AcH acetaldehyde -- DMEM Dulbecco's Modified Eagle medium -- PBS phosphate-buffered saline -- FBS fetal bovine serum -- DMSO dimethyl sulfoxide -- MTT 4, 5-dimethylthiazol-2-yl-2, 5-diphenyl tetrazolium bromide -- PI propidiumiodide -- MDC monodansyl-cadaverine -- AO acridine orange -- 3-MA 3-methyladenine -- CQ chloroquine -- AST/GOT aspartate aminotransferase -- ALT/GPT alanine aminotransferase -- HRP horseradish peroxidase -- PARP poly ADP-ribose polymerase -- Bcl-2 B-cell lymphoma-2 -- LC3 microtubule-associated protein 1 light chain 3; Toxicity testing -- In vitro -- Periodicals; Toxicology -- Periodicals; 615.9
• DOI: 10.1016/j.tiv.2020.105047
• Languages: English
• ISSNs: 0887-2333

Abstract

Abstract: Silibinin, one of the flavonoids isolated from milk thistle seeds of Silybum marianum, has hepatoprotective properties against toxins in clinical. However, the detailed mechanisms have remained unclear. This study investigates the underlying mechanism of silibinin in the protection against ethanol- or acetaldehyde-induced damage of neonatal mouse primary hepatocytes in vitro . The results show that ethanol inhibited proliferation of hepatocytes in a time (12, 24, 36 h) and dose-dependent (0–800 mM) manner. However, silibinin did not show protective effect on ethanol (500 mM)-induced suppression of hepatocyte proliferation. Acetaldehyde, the toxic metabolite of ethanol, appearing immediately in individuals after drink also inhibited the proliferation of hepatocytes in a dose-dependent (0–12 mM) manner. Surprisingly, silibinin significantly increased the cell viability and reduced the leakage of alanine amino transferase (ALT) and aspartate amino transferase (AST) in acetaldehyde-treated hepatocytes, suggesting that silibinin protected cell injury caused by acetaldehyde treatment. The apoptosis-inducing effect of acetaldehyde was demonstrated by the increased number of cells in sub-G1 phase as well as caspase-3 activation. Further study shows that acetaldehyde induced autophagy in the hepatocytes. The autophagy inhibitors, 3-Methyladenine (3-MA) and chloroquine (CQ), further decreased the viability of cells treated with acetaldehyde, suggesting that autophagy plays a protective role against apoptosis. Consistently, silibinin (20 μM) significantly reduced the activation of caspase 3 or apoptosis and increased the conversion of LC3-I to LC3-II or autophagy. Taken together, it is concluded that silibinin does not repress the ethanol- induced hepatocyte injury, whereas silibinin reduces acetaldehyde-caused hepatocyte injury through down-regulation of apoptosis and up-regulation of autophagy. Highlights: Acetaldehyde (ACH) induced apoptosis in liver cells; AcH induced autophagy in liver cells; Silibinin protects AcH-induced death, but not ethanol-induced death of hepatocytes.