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. 2023 Jun 12;28(12):4720.
doi: 10.3390/molecules28124720.

Mitigation of Hepatotoxicity via Boosting Antioxidants and Reducing Oxidative Stress and Inflammation in Carbendazim-Treated Rats Using Adiantum Capillus-Veneris L. Extract

Mohamed Seif  1 , Hanan Aati  2 , May Amer  1 , Arthur J Ragauskas  3 , Amr Seif  4 , Ahmed H El-Sappah  5 , Abdulrahman Aati  6 , Abd El-Nasser A Madboli  7 , Mahmoud Emam  8
Affiliations

Mitigation of Hepatotoxicity via Boosting Antioxidants and Reducing Oxidative Stress and Inflammation in Carbendazim-Treated Rats Using Adiantum Capillus-Veneris L. Extract

Mohamed Seif et al. Molecules. .

Abstract

Exposure to food contaminants continues to be a substantial source of human health risks all over the world, particularly in developing countries. Carbendazim (CBZ) is a chemical fungicide used to control the spread of various fungi and other pathogens in the agriculture and veterinary sectors. The hazardous effects of CBZ on human health occur due to the accumulation of its residues in agricultural food products. In this study, the possible hepatoprotective effects of Adiantum capillus-veneris L. (ACVL) extract were evaluated in CBZ-treated rats. A GC-MS analysis revealed that ACVL extract contained several bioactive hydrocarbon components and fatty acids, and that the components exerted hepatic protection by mitigating oxidative stress via upregulating antioxidant agents and neutralizing nitrogen and oxygen free radicals. Moreover, ACVL extracts relieved hepatic inflammation via decreasing NO, NF-κB, and pro-inflammatory cytokines (TNF-a, IL-6) in the liver of CBZ-treated rats, both at protein and mRNA levels. In addition, the protective effect of ACVL has appeared in the histopathological figures and function markers in the livers of CBZ-treated rats. According to the present results, ACVL extract can protect the hepatic tissue and restore its functions to a control level in CBZ-treated rats; this effect may be attributed to its antioxidant and anti-inflammatory activities.

Keywords: A. capillus-veneris L.; GC-MS; carbendazim; hepatotoxicity; protection; rats.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Total ion chromatogram (TIC) of GC-MS of the unsaponifiable matter of ACVL (A); illustration of the percentage of different classes (B).
Figure 2
Figure 2
Total ion chromatogram (TIC) of GC-MS of the fatty acids of ACVL (A); illustration of the percentage of different classes of fatty acids (B).
Figure 3
Figure 3
Change in body weight of rats treated with Adiantum Capillus-Veneris L. (ACVL) and carbendazim (CBZ) at the end of the experiment. The different letters represent statistically significant differences (p < 0.05) between treatment and control.
Figure 4
Figure 4
Liver section photomicrographs of rats in the control group and ACVL-treated rats ((A,B), respectively) exhibited normal histological structures of hepatic tissues without any pathological changes (×100). A photomicrograph of a liver section of CBZ-treated rats (C) displayed severe dilatation and congestion in the portal vein branch and blood sinusoids (black arrows-×100). Moreover, the CBZ led to focal and periportal necrosis (yellow arrow) of the hepatocytes that surrounded the portal area (yellow arrows). The liver section of ACVL + CBZ-treated rats (D) revealed recovery and an absence of congestion and hemorrhage in the hepato-portal area (×100).
Figure 5
Figure 5
Hepatic oxidative stress biomarkers in different experimental groups. Data arerepresented as mean ± SD (n = 7). ACVL = Adiantum Capillus-Veneris L. CBZ = carbendazim; H2O2 = hydrogen peroxide; MDA = malondialdehyde. The different letters represent statistically significant differences (p < 0.05) between treatment and control.
Figure 6
Figure 6
Hepatic antioxidant biomarkers at protein and gene levels in different experimental groups. Data are represented as mean ± SD (n = 7). ACVL = Adiantum Capillus-Veneris L. CBZ = carbendazim; SOD = superoxide; dismutase; CAT = catalase. The different letters represent statistically significant differences (p < 0.05) between treatment and control.
Figure 7
Figure 7
Hepatic glutathione system-related biomarkers at protein and gene levels in different experimental groups. Data are represented as mean ± SD (n = 7). ACVL = Adiantum Capillus-Veneris L. CBZ = carbendazim; GSH = glutathione; GSSG = GSH disulfide; GPx = glutathione peroxidase; GSH/GSSG = the ratio between reduced glutathione and glutathione disulfide. The different letters represent statistically significant differences (p < 0.05) between treatment and control.
Figure 8
Figure 8
Biomarkers of liver injury in different experimental groups. Data represented as mean ± SD (n = 6). ALT = alanine amino transaminase, AST = aspartate aminotransaminase, ALP alkaline phosphatase, LDH = lactate dehydrogenase; ACVL = Adiantum Capillus-Veneris L. CBZ = carbendazim. The different letters represent the statistically significant differences (p < 0.05) between treatment and control.
Figure 9
Figure 9
Hepatic inflammation biomarkers of NF-κB, TNF-α, and IL-6, of protein and gene expression in different experimental groups. Data are represented as mean ± SD (n = 6). NF-κB = NF-kappa B; TNFα = tumor necrosis factor-alpha; IL6 = interleukin; inos = inducible nitric oxide; ACVL = Adiantum Capillus-Veneris L. CBZ = carbendazim. The different letters represent statistically significant differences (p < 0.05) between treatment and control.
Figure 10
Figure 10
Immunohistochemistry photomicrographs of detection of NF-κB in the different treated groups. The slides of control and ACVL-treated groups (A,B) showed a negative immunoreaction response against NF-κB-P65 in the liver tissues. Meanwhile, the slide of the CBZ-treated group (C) showed positive brown-stained immunoreaction bands toward the NF-κB-P65 antigen (red arrows). The slide of the group treated with both ACVL + CBZ showed no immunoreaction response toward the NF-κB-P65 antigen (D).
Figure 11
Figure 11
Schematic of biological experiments. ACVL = Adiantum Capillus-Veneris L.; CBZ = Carbendazim; S.D. rats = Sprague Dawley rats; ELISA = enzyme-linked immunosorbent assay.
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