Abstract
The apoptosis activity and clinical state in vitamin and mineral supplemented male Wistar rats was evaluated after carbon tetrachloride exposure (CCL4). The animals were divided equally into 6 groups (3 control groups and 3 exposure groups) with the control groups (C-75, C-30, C-19) receiving AIN-93, a specific diet for rodents, consisting of a 75%, 30% and 19% ratio of vitamins (B1, B2, B3, B6) and minerals (Fe3+ and Mg2+) and exposure groups (E-75, E-30, E-19) receiving the same diet paradigm as with the control groups but with the additional CCL4 administered once a week as an olive oil solution (control groups received the same ratio of olive oil without CCL4) for a duration of 64 days. The systemic condition of the male Wistar rats was evaluated based on morphological parameters and hematological and biochemical analysis, whereas the apoptosis activity in the liver was evaluated via comet assay techniques. The apoptosis activity in the liver of control and exposure groups increased compared to the decrease in the essential substance provisions with the E-75 group reaching 129% (p < 0.05) higher levels compared to the C-75 group, and 98% (p < 0.05) and 23% (p > 0.05) higher in the E-30 and E-19 groups compared to the C-30 and C-19 groups, respectively. From the apoptosis results and clinical state evaluation, it is clearly demonstrated that the effectiveness of using apoptosis activity as a biomarker after CCL4 exposure and the vitamin and mineral absorption capability in male Wistar rats can be applied as an evaluating method for toxicological research.
Highlights
Vitamins and micronutrients have been extensively studied showing that when a deficiency of an essential substance exists, it can play a major role in following associated metabolic disorders [1,2]
The animals were divided and randomly into 6 groups (3 control groups and 3 exposure groups) with the control groups (C-75, C-30, C-19) receiving American Institute of Nutrition Rodent Diets (AIN)-93, a specific diet for rodents, consisting of a 75%, 30% and 19% ratio of vitamins (B1, B2, B3, B6) and minerals (Fe3+ and Mg2+) and exposure groups (E-75, E-30, E-19) receiving the same diet paradigm as with the control groups but with the additional CCL4 (≥99.5%, 289116 Sigma-Aldrich) administered once a week as an olive oil solution with
The rate of growth depended on the administered vitamins with the initial bodyweight of all groups starting at 85.6 ± 1.0 g, and ending at 403.4 ± 8.7, 378.7 ± 8.1 and 338.7 ± 7.7 g for the C-75, C-30 and C-19 groups and 373.4 ± 8.3, 352.0 ± 7.0 and 326.9 ± 8.7 g for the exposed groups, E-75, E-30 and E-19 respectively
Summary
Vitamins and micronutrients have been extensively studied showing that when a deficiency of an essential substance exists, it can play a major role in following associated metabolic disorders [1,2]. Iron is a component of cytochromes which are involved in the metabolism of xenobiotics, as well as part of catalase, a key enzyme of the antioxidant defense system [3]. Taking into account just a few of the previously described examples of vitamin and micronutrient deficiency, it is important to discover new methods to clarify which exact parameters are to blame and how are they able to induce possible adverse effects. One of the current trends in modern science is the research and development of toxicological methods, aiming to clarify hidden adverse effects of various low toxic chemical factors. Throughout the previous years, ways of obtaining such results was achieved by identifying new sensitive and specific biomarkers [7,8,9,10,11], choosing new biological objects (e.g., genetically modified organisms and organisms of synthetic biology that are highly similar to humans in their biochemical, physiological, pathological aspect of view) [12,13,14], running computer simulations [15,16,17] and by developing new toxicological models for laboratory animals [18,19,20]
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