Effect of silibinin on ethanol- or acetaldehyde-induced damge of mouse primary hepatocytes in vitro
Effect of silibinin on ethanol- or acetaldehyde-induced damge of mouse primary hepatocytes in vitro
复制标题
水飞蓟宾对乙醇或乙醛诱导的体外小鼠原代肝细胞损伤的影响
DOI：
10.1016/j.tiv.2020.105047
复制
发表时间：
2021-02-01
影响因子：
3.2
通讯作者：
Ikejima, Takashi
中科院分区：
文献类型：
--
作者：
Song, Xiao-Yu;Li, Rong-Hua;Ikejima, Takashi
水飞蓟宾是从水飞蓟种子中分离得到的黄酮类化合物，临床上对毒素具有保肝作用。然而，详细的机制仍不清楚。本研究探讨了水飞蓟宾对乙醇或乙酰丙酮诱导的新生小鼠原代肝细胞损伤的保护作用及其机制。结果表明，乙醇抑制肝细胞增殖的时间（12，24，36小时）和剂量依赖性（0 - 800 mM）的方式。然而，水飞蓟宾没有显示出对乙醇（500 mM）诱导的肝细胞增殖抑制的保护作用。乙醛，乙醇的有毒代谢产物，出现在饮酒后立即在个人也抑制肝细胞的增殖，在剂量依赖性（0 - 12毫米）的方式。令人惊讶的是，水飞蓟宾显著增加了细胞活力，并减少了乙醛处理的肝细胞中丙氨酸氨基转移酶（ALT）和天冬氨酸氨基转移酶（AST）的泄漏，表明水飞蓟宾保护了乙醛处理引起的细胞损伤。乙醛的凋亡诱导作用通过亚G1期细胞数量的增加以及caspase-3的激活来证明。进一步的研究表明，乙醛诱导肝细胞自噬。自噬抑制剂3-甲基腺嘌呤（3-MA）和氯喹（CQ）进一步降低了乙醛处理的细胞的活力，表明自噬对细胞凋亡起保护作用。一致地，水飞蓟宾（20 μ M）显著减少半胱天冬酶3的活化或凋亡，并增加LC 3-I向LC 3-II的转化或自噬。综上所述，可以得出结论，水飞蓟宾不抑制乙醇诱导的肝细胞损伤，而水飞蓟宾通过下调细胞凋亡和上调自噬来减少乙酰丙酮化物引起的肝细胞损伤。
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 mu M) 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.