Comparative antidotal efficacy of benzylpenicillin, ceftazidime and rifamycin in cultured human hepatocytes intoxicated with α-amanitin.
The most often used antidote to treat poisoning caused by amanitin-containing mushrooms is benzylpenicillin (BPCN). However, a very few reports suggest that other antibiotics such as ceftazidime (CEFT) and rifamycin SV (RIFSV) show better antidote activity against amanitins than BPCN. Given this, there is an ongoing debate as which of three antidotes is optimal for treatment of such poisonings. In this study, the efficacy of BPCN was compared with those of CEFT and RIFSV in human hepatocyte model. The functional integrity and viability of cultured hepatocytes was evaluated using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay and by measurements of lactic dehydrogenase (LDH) activity. In the first experimental layout, hepatocytes were simultaneously exposed to α-AMA and tested antidotes, while in the second layout, the cells were exposed for the first 12 h to α-AMA only, and then, the medium containing α-AMA was exchanged to culture medium containing both α-AMA and the antidotes tested. The results demonstrated that simultaneous administration of α-AMA and each of tested antidotes (BPCN, CEFT, RIFSV) effectively protected human hepatocytes; however, in the group dosed with BPCN, the highest hepatocyte viability was observed. In cell cultures from experimental layout II, all tested antidotes were ineffective, which indicates that after the critical dose of α-AMA had been taken up by hepatocytes, further suppression of this process does not protect the cells against injury. Thus, 12 h of exposure of incubated hepatocytes to α-AMA is a sufficient time for such a cellular uptake of a critical dose of this toxin. In summary, it can be concluded that easily accessible and low-cost BPCN should be widely used as an antidote against amanitins. However, the key to successful therapy is a quick implementation of an antidote in order to protect as large as possible portion of the liver parenchyma against the devastating uptake of a critical dose of amanitins.
Magdalan
Jan
J
Department of Pharmacology, Wrociaw Medical University, Mikulicza-Radeckiego 2, 50-345 Wrociaw, Poland.
Ostrowska
Alina
A
Piotrowska
Aleksandra
A
Gomułkiewicz
Agnieszka
A
Szeląg
Adam
A
Dziędgiel
Piotr
P
D053959:Alpha-Amanitin / Q000633:toxicity*; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D000931:Antidotes / Q000494:pharmacology*; D002442:Ceftazidime / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D007770:L-Lactate Dehydrogenase; D009145:Mushroom Poisoning / Q000188:drug therapy; D010400:Penicillin G / Q000494:pharmacology*; D012294:Rifamycins / Q000494:pharmacology*; D013778:Tetrazolium Salts / Q000378:metabolism; D013844:Thiazoles / Q000378:metabolism
D053959:Alpha-Amanitin; D000900:Anti-Bacterial Agents; D000931:Antidotes; D012294:Rifamycins; D013778:Tetrazolium Salts; D013844:Thiazoles; D002442:Ceftazidime; C023808:rifamycin SV; D007770:L-Lactate Dehydrogenase; C022616:thiazolyl blue; D010400:Penicillin G