Protective effects of a bioactive antioxidant complex against Aspartame exposure during gestation. Biochemical, morphological and ultrastructural studies on new-born rat brain

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Article Title:	Protective effects of a bioactive antioxidant complex against Aspartame exposure during gestation. Biochemical, morphological and ultrastructural studies on new-born rat brain
Authors:	Constantin Puică, Constantin Crăciun, Maria Borsa, Mircea Rusu, Ioana Roman, Mihai Cristescu
Affiliation:	1 Institute of Biological Research Cluj-Napoca, 2 University Babes-Bolyai Cluj-Napoca, 3 University of Agricultural Sciences and Veterinary Medicine, Cluj-Napoca
Abstract:	90 days-old nuliparous female Wistar rats were treated by intragastrical gavages with Aspartame at a dose of 0.007 mg/g. bw, dissolved in distilled water (A group), during the entire period of gestation. The animals of group AC were daily treated by oral gavages with Aspartame in each dose, associate with vitamin C in a dose of 0,01 mg/g bw, vitamin E in a dose of 0.002 mg/g bw, and selenium in a dose of 0.005 μg/g bw dissolved in distilled water during the entire period of gestation (days 1-20 of gestation). The new-born brains were examined for their ultrastructure. The rate of some oxidative stress parameters (lipoperoxidation-LPO, non-protein SH and total SH-protein levels) from brain homogenate, were also estimated. The new-born rats prenatally treated with Aspartame showed significant increase of the LPO rate in the brain homogenate (1.17 0.08 nmol MDA/h; +19.3%; p ≤ 0.05), compared to 0.98 ± 0.04 nmol MDA/h value registered in control group. The administration of the bioactive mixture (AC group) showed a moderate, non-significant increase of LPO content in new-born brain homogenate, (1.01  0.05 nmol MDA/h; +3.06%), compared to LPO level in control group. Aspartame treatment during gestation induced a significant depletion of total SH groups in the brain homogenate (14.5 ± 0.27 μmol/g cerebral tissue, – 6.45%, p < 0.5), compared to controls (15.5 ± 0.32 μmol/g). Moderate, but significant decrease of total SH groups was registered in AC group: (14.9 ± 0.25 μmol/g; – 9.61%; p < 0.01). Significant depletion the nonprotein SH (new-born brain glutathione content) was registered following Aspartame treatment during gestation: (4.95 ± 0.32 μmol/g. -13.15%; p < 0.001), compared to 5.70 ± 0.32 μmol/g. value registered in controls, and moderate and no significant difference of non-protein SH content, compared to controls, was observed in AC group: (5.48 ± 0.29; – 3.85%; p > 0.25). The study of new-born brain ultrastructure revealed that treatment with Aspartame during gestation has induced decreases with different intensity degrees of both size of neurons and their nuclei in all brain areas. Some nuclei are picnotic. Degenerative aspects of all cellular constituents are typical features of apoptosis: irregular outline of nuclei; picnotic and intense electron dense nuclei; mitochondrial and rough endoplasmic reticulum swellings; demyelization of the myelin sheath; intense vacuolization both of the cytoplasm and axoplasma. The administration of vitamins C, E and selenium produced moderate protective effects against Aspartame treatment, characterized by a some reduction of the lesion-necrotic aspects of the neurons and of all cellular organelles. Our images show groups of neurons with an aspect close to normal of the nuclei and organelles. Ultrastructural and biochemical studies represents an additional element for the establishing the direct embryotoxic/neurotoxic effect of Aspartame-treatment on new-born brain structures and functionality. At the same time, moderate, but positive modulation of the bioactive compound on the ultrastructure and functionality of these brain parameters was registered.
Keywords:	protective, antioxidant, Aspartame, gestation, brain fetuses
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*Correspondence:	Dr. Puică Constantin, “Institutul de Cercetari Biologice Cluj-Napoca”, str. Republicii nr. 48, cod 400015 Tel./Fax +40-(0264)-591238, email: ctin_puica@yahoo.com