J Korean Soc Clin Toxicol.  2019 Dec;17(2):58-65. 10.22537/jksct.2019.17.2.58.

The Protective Effect of Green Tea Extract on Alpha-amanitin Induced Hepatotoxicity

Affiliations
  • 1Department of Emergency Medicine, School of Medicine, Chosun University, Gwangju, Korea. eryongjin@chosun.ac.kr
  • 2Department of Pathology, School of Medicine, Chosun University, Gwangju, Korea.
  • 3Department of Biochemistry, School of Medicine, Chosun University, Gwangju, Korea.

Abstract

PURPOSE
Alpha-amanitin induces potent oxidative stress and apoptosis, and may play a significant role in the pathogenesis of hepatotoxicity. This study examined the mechanisms of α-amanitin-induced apoptosis in vitro, and whether green tea extract (GTE) offers protection against hepatic damage caused by α-amanitin (AMA) induced apoptosis in vivo.
METHODS
The effects of GTE and SIL on the cell viability of cultured murine hepatocytes induced by AMA were evaluated using an MTT assay. Apoptosis was assessed by an analysis of DNA fragmentation and caspase-3. In the in vivo protocol, mice were divided into the following four groups: control group (0.9% saline injection), AMA group (α-amanitin 0.6 mg/kg), AMA+SIL group (α-amanitin and silibinin 50 mg/kg), and AMA+GTE group (α-amanitin and green tea extract 25 mg/kg). After 48 hours of treatment, the hepatic aminotransferase and the extent of hepatonecrosis of each subject was evaluated.
RESULTS
In the hepatocytes exposed to AMA and the tested antidotes, the cell viability was significantly lower than the AMA only group. An analysis of DNA fragmentation showed distinctive cleavage of hepatocyte nuclear DNA in the cells exposed to AMA. In addition, the AMA and GTE or SIL groups showed more relief of the cleavage of the nuclear DNA ladder. Similarly, values of caspase-3 in the AMA+GTE and AMA+SIL groups were significantly lower than in the AMA group. The serum AST and ALT levels were significantly higher in the AMA group than in the control and significantly lower in the AMA+GTE group. In addition, AMA+GTE induced a significant decrease in hepatonecrosis compared to the controls when a histologic grading scale was used.
CONCLUSION
GTE is effective against AMA-induced hepatotoxicity with its apoptosis regulatory properties under in vitro and in vivo conditions.

Keyword

Alpha-amanitin; Green tea extract; Liver toxicity; Antidotes; Apoptosis

MeSH Terms

Alpha-Amanitin*
Animals
Antidotes
Apoptosis
Caspase 3
Cell Survival
DNA
DNA Fragmentation
Hepatocytes
In Vitro Techniques
Mice
Oxidative Stress
Tea*
Alpha-Amanitin
Antidotes
Caspase 3
DNA
Tea
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