Protective effects of silibinin against ethanol- or acetaldehyde-caused damage in liver cell lines involve the repression of mitochondrial fission. (April 2022)

• Record Type: Journal Article
• Authors: Song, Xiao-Yu; Liu, Peng-Cheng; Liu, Wei-Wei; Hayashi, Toshihiko; Mizuno, Kazunori; Hattori, Shunji; Fujisaki, Hitomi; Ikejima, Takashi
• Is Part Of: Toxicology in vitro. Volume 80(2022)
• Publication Date: 2022-04
• Subjects: Ethanol -- Acetaldehyde -- Silibinin -- Apoptosis -- Mitochondrial fission; HPLC High performance liquid chromatography -- DMEM Dulbecco's Modified Eagle medium -- Alde Acetaldehyde -- Eth Ethanol -- Sili Silibinin -- PBS Phosphate-buffered saline -- FBS Fetal bovine serum -- DMSO Dimethyl sulfoxide -- MTT 4, 5-dimethylthiazol-2-yl-2, 5-diphenyl tetrazolium bromide -- AST Aspartate aminotransferase -- ALT Alanine aminotransferase -- HRP Horseradish peroxidase -- PARP Poly ADP-ribose polymerase -- Bcl-2 B-cell lymphoma-2 -- Bax BCL2-Associated X -- DRP1 Dynamin-related protein 1 -- OPA1 Optic atrophy 1 -- MFN1 Mitofusin 1 -- STS Staurosporine -- Mdivi-1 Mitochondrial division inhibitor 1 -- ALD Alcoholic liver disease -- ALDH aldehyde dehydrogenase 2 family -- ADH Alcohol dehydrogenase -- CYP2E1 Cytochrome P450 2E1 -- ATP Adenosine triphosphate -- ROS Reactive oxygen species -- H2DCF-DA 2′, 7′-dichlorodihydrofluorescein diacetate -- MDA Malondialdehyde -- MTG MitoTracker Green -- MTR MitoTracker Deep Red FM.; Toxicity testing -- In vitro -- Periodicals; Toxicology -- Periodicals; 615.9
• DOI: 10.1016/j.tiv.2022.105330
• Languages: English
• ISSNs: 0887-2333

Abstract

Abstract: Silibinin is a natural polyphenolic flavonoid, isolated from the seeds of the milk thistle of Silybum marianum (L.) Gaertn . Silibinin has been widely used clinically as a traditional medicine for liver diseases. This study investigated the protective role of silibinin in ethanol- or acetaldehyde-induced apoptosis in human carcinomatous liver HepG2 cells and immortalized liver HL7702 cells, focusing on elucidation of the underlying mechanism in vitro. The toxicity of ethanol or acetaldehyde was evaluated by MTT assay. Apoptosis-related proteins, mitochondrial fission-associated proteins and mitochondrial fusion-associated proteins were analyzed by western blotting and immunofluorescence microscopy. Present experimental results demonstrated that silibinin improved cell viability, reduced the enzyme activities of AST/ALT and ALDH/ADH, inhibited apoptosis and recovered mitochondrial function in ethanol- or acetaldehyde-treated HepG2 or HL7702 cells. Silibinin reduced the expression of mitochondrial fission-associated proteins, dynamin-related protein 1 (DRP1), but increased mitochondrial fusion-associated proteins, optic atrophy 1 (OPA1) and mitofusin 1 (MFN1). Accordingly, inhibition of DRP1 activity with its pharmacological inhibitor or siDRP1 efficiently attenuated ethanol- or acetaldehyde-induced apoptosis, whereas activation of DRP1 by using staurosporine (STS) further increased apoptosis in ethanol- or acetaldehyde-treated HepG2 or HL7702 cells. The results show that silibinin protects cells against ethanol- or acetaldehyde-induced mitochondrial fission that results in apoptosis. Highlights: Ethanol or acetaldehyde induces apoptosis in HepG2 and HL7702 cells. Silibinin reduces ethanol- or acetaldehyde-induced apoptosis. Ethanol or acetaldehyde aggravates mitochondrial fission resulting in apoptosis. Silibinin restores ethanol- or acetaldehyde-caused mitochondrial disorders.