Effects of Gallic Acid on Antioxidant Defense System in Mice with Benzene-Induced Myelotoxicity

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Abstract

Benzene is known to cause myelotoxicity which impacts secondary complications mediated by oxidative stress on the heart and erythrocyte. Gallic acid is an antioxidant which has not been evaluated for protective effect against cardiovascular complications of benzene-induced myelotoxicity. Therefore, this study was carried out to evaluate the effects of gallic acid on the concentrations of selected oxidative stress biomarkers and activities of antioxidant enzymes in the erythrocyte and heart of mice with benzene-induced myelotoxicity. Thirty-six male mice were randomized into six groups of six mice each. Group A served as the normal control receiving distilled water while the remaining groups were orally administered 150 mg/kg body weight of benzene for fourteen days. Distilled water, 50 mg/kg body weight of ascorbic acid, 25, 50, and 100 mg/kg body weight of gallic acid were simultaneously administered to mice of groups B (negative control), C, D, E, and F respectively for fourteen days. Concentrations of oxidative stress biomarkers and activities of antioxidant enzymes in target tissues were then determined. Results revealed significant elevation (p<0.05) in the concentrations of malondialdehyde, nitric oxide and protein carbonyl, as well as significant decrease (p<0.05) in the concentrations of reduced glutathione, protein, and activities of catalase, glutathione peroxidase, glutathione-S-transferase and superoxide dismutase in the heart and erythrocyte of negative control compared to normal control. However, treatment with gallic acid at various doses significantly reverted (p<0.05) the observed alterations in these parameters, comparing favourably with ascorbic acid (reference drug). These results suggest that gallic acid protected the heart and erythrocyte against lipid and protein oxidation, and enhanced antioxidant defense system of mice with benzene-induced myelotoxicity. Keywords: myelotoxicity; oxidative stress; gallic acid; ascorbic acid; benzene; antioxidant enzymes; biomarkers; lipid peroxidation; protein oxidation.

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    This study was designed to investigate the effects of gallic acid on the concentrations of selected oxidative stress biomarkers and activities of antioxidant enzymes in the erythrocyte and heart of mice after benzene administration.  The authors conclude that gallic acid protected the heart and erythrocyte against lipid and protein oxidation, and enhanced the antioxidant defense system of mice with benzene.

    Major issues

    -          In the title, the authors identify that they will examine the effect of gallic acid in Benzene-induced myelotoxicity, however, the myelotoxicity was not investigated all over the study the bone marrow was not investigated.

    -          It is not clear why authors chose heart only to assess oxidative stress activity as a result of benzene administration

    -          In the introduction section: background on gallic acid's nature, source, and Pharmacological effects motivate the authors to study its effect. The following references could help:

    • Kahkeshani, N., Farzaei, F., Fotouhi, M., Alavi, S. S., Bahramsoltani, R., Naseri, R., Momtaz, S., Abbasabadi, Z., Rahimi, R., Farzaei, M. H., & Bishayee, A. (2019). Pharmacological effects of gallic acid in health and diseases: A mechanistic review. Iranian journal of basic medical sciences, 22(3), 225–237. https://doi.org/10.22038/ijbms.2019.32806.7897.

    • Jasemi, S. V., Khazaei, H., Momtaz, S., Farzaei, M. H., & Echeverría, J. (2022). Natural products in the treatment of pulmonary emphysema: Therapeutic effects and mechanisms of action. Phytomedicine, 99, 153988. https://doi.org/10.1016/j.phymed.2022.153988

      -          In the Blood sample collection section: Please identify how the authors collect venous blood samples (retro-orbital, tail vein, or identify your method)

      -          In the Blood sample preparation section: How do the authors separate RBCs from the other cells in the buffy coat and what is the benefit of this preparation (please add reference)

      -          In the Red blood cell lysis section: Please explain if RBC lysis was done by adding lysis buffer to 1 ml of blood or packed RBCs. Please add a reference to the technique

      -          In the Protein concentration section (procedures): Please identify how the authors collected the protein sample in which the protein concentration is measured

      -          In the Catalase (CAT) section (procedures): Please identify the sample in which the catalase is measured

      -          Please explain which protein sample that used to assess Glutathione-S-transferase (GST) So the unit is expressed as mmol of CNDB formed/min/mg protein

      -          In the results section: In all figures, the bar charts are signed with (a, b&c)letters but the authors did not define what these signs mean especially since the control group is also signed. Please revise and explain

      -          The 1st paragraph of the discussion is previously mentioned in the introduction section (repeated data)

    Minor issues

    -          In the introduction section: Kippen et al., 1988, This reference is not included in the reference section

    -          In the Preparation of tissue homogenate section: please add a reference to the technique of preparation of heart tissue homogenate.

    -          In the results section: The authors said, "comparing favorably well with ascorbic acid". Please be more specific by expressing the comparison scientifically

    -          In the reference section: the references should be arranged alphabetically

    -          Reference 27: is not cited in the article

    -          Reference 36: the correct name of the 1st author is Gutteridge

    -          Reference 37 is a repetition of reference 18

    -          Reference 42 is a repetition of reference 36

    Competing interests

    The author declares that they have no competing interests.