BIODETOXIFICATION OF AFLATOXIN B1 USING PURE COMPOUNDS FROM Achyrocline satureioides

ESCOBAR FRANCO MATÍAS; MAGNOLI ALEJANDRA; OLIVA MARÍA DE LAS MERCEDES; SABINI MARÍA CAROLA; MENIS CANDELA FLORENCIA; SORIA ELIO ANDRÉS; CAMPRA NOELIA; CARIDDI LAURA NOELIA

Mendoza

Congreso; XXXVI REUNIÓN CIENTIFICA ANUAL DE LA SOCIEDAD DE BIOLOGÍA DE CUYO; 2018

Sociedad de Biología de Cuyo

Aflatoxin B1 (AFB1), produced by strains of Aspergillus flavus and A. parasiticus, is carcinogenic, teratogenic, hepatotoxic and immunotoxic and, at high doses, can be lethal. Also, fungal toxins are important because they generate great economic losses. Natural products have been shown to have detoxifying capacity of mycotoxins. The objective of this work was to evaluate in vivo, by oral administration in Wistar rats, the ability of luteolin (L) and chlorogenic acid (CHL) to attenuate or revert cytotoxic and genotoxic effect of AFB1 by supplying the toxin for 30 days. For this experience Wistar rats (Rattus norvegicus albinas) of 200 g of body weight were used, which were divided into 6 groups of 4 individuals each one. They were orally administered 0.2 ml of the different treatments. AFB1 was dissolved in 0.05% of dimethyl sulfoxide (DMSO) and 0.1 M of sodium bicarbonate (pH 7.4). The compounds were administered at non-cytogenotoxic concentrations by gastric tube. Groups: Control AFB1 (400 ppb); Treatment 1: AFB1 (400 ppb) + L (0.5 mg/kg b.w.); Treatment 2: AFB1 (400 ppb) + CHL (5 mg/kg b.w.); Control drug 1: L (0.5 mg/kg b.w.); Control drug 2: CHL (5 mg/kg b.w.); Negative Control: 0.1 M sodium bicarbonate. In all cases they were administered water and food ad libitum. The treatment was extended for 30 days. The detoxifying effects were evaluated by: 1. Evaluation of genotoxicity and antigenotoxicity by Micronucleus test in rat bone marrow: from the femurs of rats, the bone marrow samples were obtained with fetal calf serum, were homogenized, centrifuged, and plated on slides which were fixed by soft flutter and stained with May-Grünwald and Giemsa. The frequency of micronuclei in 1000 polychromatic erythrocytes (PCE) was determined per slide. Cytotoxic and anticytotoxic effects were evaluated through the determination of the toxicity index (TI): polychromatic erythrocytes (PCE)/normochromatic erythrocytes (NCE) by 1000 PCE counted. 2. Studies of oxidative stress and antioxidant action: it was performed by determining substances reactive to thiobarbituric acid (TBARs) from liver and kidney samples. The determination of TBARs was carried out using malondialdehyde (MDA) as a reference substance. To the homogenates obtained in phosphate buffer, BHT was added, the samples were centrifuged, and the supernatant was incubated at 90°C for 30 min with thiobarbituric acid (TBA, 0.375%). To calculate MDA concentrations in the tissues, a calibration curve was used. The tests were carried out in duplicate and each sample was read in triplicate. The results are expressed as nmol of MDA/g of tissue. The results obtained indicated a significant increase in the genotoxicity index in rats treated with AFB1 (20 ± 1 MN/1000 PCE) respect to NC (6 ± 0.5). Treatment with L and CHL managed to reverse/protect from damage induced by AFB1, both, L + AFB1 and CHL + AFB1, indicated a normal value of MN/1000 PCE. On the other hand, TI showed no significant difference between any of the treatments. With regard to studies of oxidative stress, it was observed that there was an increase in TBARs in livers of rats treated with AFB1 in relation to NC but without statistical significance. On the other hand, kidneys showed a significant increase in TBARs in rats treated with AFB1 respect to NC. Treatments with L and CHL protected from damage induced by AFB1.