Gene expression profiles for apoptotic and necrotic pathways during Amanita phalloides intoxication in mice

Year 2022, Volume: 52 Issue: 3, 281 - 288, 30.12.2022

Selim Karahan Zehra Atli Ertuğrul Kaya Feride Özdemir Mehmet Boğa Sevcan İzgi

https://doi.org/10.26650/IstanbulJPharm.2022.1136288

Abstract

Background and Aims: Amanita phalloides is the deadliest toxic mushroom in the world and causes death from acute liver failure. α-amanitin (α-AMA), the most potent toxin, inhibits RNA polymerase II in hepatocytes, stops protein synthesis, and causes hepatotoxicity. However, the information about the mechanisms underlying hepatotoxicity caused by α-AMA is quite inadequate. This study aims to reveal the complex necrotic and apoptotic mechanisms occurring in mouse hepatocytes de- pending on A. phalloides exposure time in vivo.
Methods: BALB-c male mice were divided into 5 groups (n=7): control, α-AMA-2, α-AMA-12, α-AMA-72, and α-AMA-96 groups. A poisoning model was created by oral administration of A. phalloides mushroom extract containing 10 mg/kg of α-AMA to mice and they were sacrificed after 2, 12, 72, and 96 h. Then, TNF-α, Bax, caspase-3, and Bcl-2 gene expression levels in liver tissues were examined by the RT-qPCR method. Time-dependent damage to liver tissues was also evaluated histopathologically.
Results: RT-qPCR results showed that proinflammatory cytokine TNF-α mRNA expression levels increased in mouse liver tissues at 2 and 12 h after A. phalloides administration compared among the groups. Bax mRNA expression levels increased in the 12 and 72 h after A. phalloides ingestion. It was observed that caspase-3 mRNA expression levels increased in the 72 and 96 h groups compared among the groups, while Bcl-2 mRNA expression levels decreased in the 72 and 96 h groups.
Conclusion: Our findings showed that necrotic mechanisms develop in the early period after A. phalloides mushroom poison- ing, and then apoptotic mechanisms are effective. In conclusion, understanding the mechanisms of A. phalloides-induced hepatotoxicity will provide important information for new treatment strategies to be developed.

Keywords

α-amanitin, TNF-α, Bax, caspase-3, Bcl-2, RT-qPCR

Supporting Institution

Scientific Research Projects Coordinator of Dicle University

Project Number

DUBAP:DUSAM.21.002

Thanks

The authors are thankful to the Scientific Research Projects Coordinator of Dicle University (DUBAP:DUSAM.21.002) for the partial supports.

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