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  • Icaritin ameliorates carbon tetrachloride-induced acute liver injury mainly because of the antioxidative function through estrogen-like effects.

Icaritin ameliorates carbon tetrachloride-induced acute liver injury mainly because of the antioxidative function through estrogen-like effects.

In vitro cellular & developmental biology. Animal (2014-08-26)
Peng Liu, Xiang Jin, Hao Lv, Jing Li, Wen Xu, Hai-hua Qian, Zhengfeng Yin
ABSTRACT

To investigate the effects of icaritin, an active ingredient extracted from Epimedium Sagittatum (Sieb. et Zucc.), on CCl4-induced liver injury and its possible mechanisms. Hepatocytes isolated from Sprague-Dawley male rats were treated with 3 mmol/L CCl4 for 24 h to induce acute liver cell injury, then icaritin (0.1, 1, 10, 100 μmol/L, respectively) was administrated to the cells, and estrogen receptor antagonist ICI182,780 (1 μmol/L) was co-treated with 10 μmol/L icaritin. Biochemical parameters (alanine aminotransferase (ALT), aspartate aminotransferase (AST), malondialdehyde (MDA), and superoxide dismutase (SOD)) and cell apoptosis were detected to evaluate the injury degree. Protein expressions of Bax, Bcl-2, liver fatty acid-binding protein (L-FABP), and peroxisome proliferator-activated receptor-α (PPAR-α) as well as reactive oxygen species (ROS) generation were determined by western blot. Icaritin alleviated CCl4-induced liver cell injury in a concentration-dependent manner and 10 μmol/L was the optimal concentration. Icaritin (10 μmol/L) significantly reduced activities of ALT, AST in cell culture medium and MDA level of the impaired liver cells, but increased the intercellular SOD activity. The apoptotic rate of the impaired liver cells was also decreased by icaritin (10 μmol/L) treatment. Icaritin might exert antioxidative and anti-apoptotic functions via estrogen-like effect, as the ratio of Bcl-2/Bax was significantly increased, while protein expressions of L-FABP and PPAR-α were markedly increased, and this function was blocked by the estrogen receptor antagonist ICI182,780 efficiently. Icaritin may be a promising drug candidate for acute liver injury benefiting from the antioxidative and anti-apoptotic functions via estrogen-like effect.

MATERIALS
Product Number
Brand
Product Description

Supelco
Carbon tetrachloride, analytical standard
Sigma-Aldrich
Carbon tetrachloride, anhydrous, ≥99.5%
Sigma-Aldrich
2-Thiobarbituric acid, ≥98%
Sigma-Aldrich
DL-Glyceraldehyde 3-phosphate solution, 45-55 mg/mL in H2O
Silybin, mixture of Silybin A and B, primary reference standard
Sigma-Aldrich
Icaritin, ≥98% (HPLC)
Sigma-Aldrich
Carbon tetrachloride, reagent grade, 99.9%
Supelco
Carbon tetrachloride, suitable for HPLC, ≥99.9%
Sigma-Aldrich
1,1,3,3-Tetraethoxypropane, ≥96%
Sigma-Aldrich
Silibinin, ≥98% (HPLC)