An approach to evaluating the potential teratogenic and neurotoxic mechanism of BHA based on apoptosis induced by oxidative stress in zebrafish embryo (Danio rerio)

Abstract

Butylated hydroxyanisole (BHA) has been widely used in the cosmetics, pharmaceutical, and food industries due to its antioxidant activity. Despite the antioxidant effects, reported adverse effects of BHA at the cellular level have made its use controversial. In this regard, this study was performed to elucidate the potential toxicity mechanism caused by BHA at the molecular level in zebrafish embryos. For this purpose, zebrafish embryos were exposed to BHA at levels of 0.5, 1, 5, 7.5 and 10 ppm and monitored at 24, 48, 72 and 96 hours. Survival rate, hatching rate and malformations were evaluated. We examined the potential for reactive oxygen species (ROS) production and apoptosis signalling accumulation in the whole body. Moreover, we evaluated histopathological and immunohistochemical (8-OHDG) characterization of the brain in zebrafish embryos at the 96th hour. We also examined apoptosis, histopathological and immunohistochemical (8-OHDG) characteristics in 96 hpf zebrafish larvae exposed to tertiary butylhydroquinone (TBHQ), one of the major metabolites of BHA, at doses of 0.5, 2.5, 3.75 and 5 ppm. Consequently, it has been considered that increased embryonic and larval malformations in this study may have been caused by ROS-induced apoptosis. After 96 h of exposure, positive 8-OHdG immunofluorescence, degenerative changes, and necrosis were observed in the brain of BHA and TBHQ-treated zebrafish larvae in a dose-dependent manner. BHA and TBHQ exposure could lead to an increase in 8-OHdG activities by resulting oxidative DNA damage. In particular, the obtained data indicate that the induction of ROS formation, occurring during exposure to BHA and/or multiple hydroxyl groups, could be responsible for apoptosis.

Algerian Bunium incrassatum Seeds: Effects of Extraction Solvent Polarity on Phenolic Profile and Antioxidant Activity

“…This may explain the high antioxidant efficiency of bioactive compounds that petroleum ether extract contains as compared to the reference compound (BHA), which is a strong synthetic antioxidant. But as usual for synthetic additives, BHA has been accepted as possibly carcinogenic to humans by the International Agency for Research on Cancer (IARC) due to the toxicological data on its mechanism of action (36). It should also be noted that all identified compounds have been reported previously in several studies performed on different species of Bunium (25,37,38), but none of them was about the seeds of B.incrassatum.…”

Terpenoids and Polyphenols as Natural Antioxidant Agents in Food Preservation

“…Although these synthetic antioxidants are widespread and strictly regulated, there are safety concerns due to their overuse and/or misuse (e.g., a mixture of certain antioxidants may increase their toxic effects) [ 23 , 24 ]. An increasing number of studies conclude that high doses of chemical synthetic antioxidants may cause DNA damage [ 25 , 26 , 27 ] or in vitro toxicity in certain tissues [ 28 , 29 ]. BHA functions as an environmental hormone and has a role as an endocrine disruptor.…”

Effects of the food colorant carmoisine on zebrafish embryos at a wide range of concentrations

“…After incubation, they were washed three times with ultra-pure H 2 O and examined under fluorescence microscopy (Zeiss, Discovery V12, Germany). Fluorescence intensity analysis of the obtained images was done with the ImageJ software (Baran et al 2020;Türkoğlu et al 2021).…”