Ameliorative effect of licorice extract against the detrimental effect of glyphosate-based pesticide: Toxicity and health

Abstract

This study sheds the light on the potential of licorice (Glycyrrhiza glabra) root aqueous extract as a cornerstone for mitigating and detoxifying the residues of the widely used agricultural Glyphosate-based pesticides (GBPs). This study examined the GBPs toxic effects on kidney, liver, thyroid functions, and apoptosis using 50 adult male albino rats. All rats were divided into 5 groups, with 10 each. Control: served as untreated rats. GBP: rats were treated with 1 mL glyphosate solution 24 % orally for three weeks. The glyphosate-treated rats were gavaged with licorice root aqueous extractsolution (100, 200, and 300 mg/mLdistilled water, respectively) daily for three weeks. Licorice root aqueous extract solution (300 mg/mL distilled water) yielded notable reductions in liver, kidney enzymes, albumin, and AFP levels within the serum. Immunological tests, including immunohistochemical evaluations of caspase-3 and TNF-α expressions revealed a dose-dependent attenuation of apoptosis and inflammation with licorice intervention. This will provide a valuable perspective for agricultural practices future and paving the way for a more sustainable approach for using GBPs in animal agriculture industries.

Keywords: Antioxidant; Apoptosis; Glyphosate-based pesticide; Licorice (Glycyrrhiza glabra); Toxicity.

Fig. 1

The effect of licorice administrationon GBP treated rats'liver functions, (A, ALT (U/L)), (B, AST (U/L)), (C, AFP (ng/mL))(D,albumin (g/dL)), within 1st, 2nd, and3^rd week of experiment. Capital letters are significance of each group vs control negative group within that time point, and small letters are significance of each treatment in week 2 and week 3 vs week 1. Control group: Rats served as controls by receiving water daily and being left untreated. GBP group: The animals were administered glyphosate solution 24 % (1 mL) orally directly into the buccal cavity of rats by gavage daily for three weeks. The glyphosate-treated animals in (GBP + GGE100), (GBP + GGE200), and (GBP + GGE300) were gavaged with licorice solution (100, 200, and 300 mg/mLDW, respectively) daily for three weeks.

Fig. 2

The effect of licorice administrationon GBP treated rats'thyroid hormones and kidney functions, (A, FT3 (ng/dL)), (B, T3 (ng/dL)), (C, T4 (μg/dL))(D, FT4 (Pg/dL)), (E, TSH ((μU/mL)(F, creatinine (mg/dL)) and (G, urea (mg/dL)), 1st, 2nd, 3rd week of experiment. Capital letters are significance of each group vs control negative group within that time point, and small letters are significance of each treatment in week 2 and week 3 vs week 1Control group: Rats served as controls by receiving water daily and being left untreated. GBP group: The animals were administered glyphosate solution 24 % (1 mL) orally directly into the buccal cavity of rats by gavage daily for three weeks. The glyphosate-treated animals in (GBP + GGE100), (GBP + GGE200), and (GBP + GGE300) were gavaged with licorice solution (100, 200, and 300 mg/mLDW, respectively) daily for three weeks.

Fig. 3

Histopathological sections of rats' livers exposed to a sub-lethal dose of glyphosate -pesticide and treated with licorice supplements. A illustrates the liver of the control group, demonstrating a normal histological appearance characterized by the presence of normal hepatic cells (black arrow) and blood sinusoids (red arrow). B, liver sections from animals treated with glyphosate display noticeable features such as swelling, vacuolation, and hepatocyte damage (yellow arrow). Panels C–E further demonstrate the liver sections of Licorice-treated animals at lower, middle, and high doses, highlighting a dose-dependent decrease in the severity of pathological alterations; however, some degenerative changes as hydropic changes could be seenafter 1st week. F Illustrates the liver of the control group, demonstrating a normal histological appearance characterized by radially arranged polyhedral hepatocytes forming cords around the central vein and sinusoids. G liver sections from animals treated with glyphosate display noticeable necrosis of centrilobular hepatocytes, dissolution (green arrow), and severe hepatocyte damage (yellow arrow). Panels H-Jfurther demonstrates the liver sections of Licorice-treated animals at lower, middle, and high doses, highlighting the approximate regain of the normal appearance of hepatocytes after 2nd week.K illustrates the liver of the control group, demonstrating a normal histological appearance characterized by the presence of Kupffer cells (blue arrow), central vein (white arrow), hepatic cells, and blood sinusoids. L liver sections from animals treated with glyphosate display noticeable features such as pyknotic nuclei (black arrow), necrosis (red arrow), dilatation in blood sinusoids, activated Kupffer cells, and fatty change. Panels M − O further demonstrates the liver sections of Licorice-treated animals at lower, middle, and high doses, highlighting a decrease in the severity of pathological alterations; however, some degenerative changes as fatty change (yellow arrow) and hydropic degeneration (green arrow), could be seen after 3rd week of experiment.

Fig. 4

Cleaved caspase-3 Immunoreactivity in Rat Liver Sections (A–F). A Illustrates the absence of cleaved caspase-3 in the Control group. Immunohistochemical analysis of TNF-α Expression in Rat Liver Sections. Panel a depicts minimal TNF-α immunoreactivity in the Control group. Panel b shows pronounced hepatic TNF-α expression in the GBP group post-glyphosate treatment. B Following glyphosate treatment, pronounced cleaved caspase-3 expression was demonstrated in the GBP group. C Depicts strong immune reactivity in liver tissues from (GBP + GGE100).D and E represent liver sections from (GBP + GGE200), and (GBP + GGE300), showing a gradient from moderate to weak cleaved caspase-3 expression. F Quantifies the immunostaining area (%) of cleaved caspase-3 expression, with data presented as mean ± SE; a distinct asterisk indicates significant differences. Arrows highlight the specific areas of immune expression. Immunohistochemicalanalysis of TNF-α Expression in Rat Liver Sections (G–L). Panel Gdepicts minimal TNF-α immunoreactivity in the Control group. Panel H shows pronounced hepatic TNF-α expression in the GBP group post-glyphosate treatment. Panels I, J, and K represent liver tissues from (GBP + GGE100), (GBP + GGE200), and (GBP + GGE300), respectively, illustrating a gradient from moderate to weak TNF-α expression. For comparative purposes, Panel L presents the quantification of the immunostaining area (%) for cleaved caspase-3 expression, with data expressed as the mean ± standard error (SE) and distinct superscript asterisk denoting statistically significant differences. Brown staining denotes TNF-α positivity.

Fig. 5

(A) Molecular docking interaction of glyphosate with rats' albumin. The Molecular docking score is −5.81 kcal/mol. Glyphosate interacted by H-donor bonds with GLU177 (4) and GLU316 and H-acceptor bonds with LYS223 (2) and ARG219(4) residues in the albumin's binding site. (B) Molecular docking interaction of glyphosate with rats' thyroxine-binding globulin. The molecular docking score is −4.73 kcal/mol. Glyphosate interacted by H-donor bonds with SER121 and LEU129 and H-acceptor bonds with ARG402, ASN126, and ASN267 residues in the thyroxine-binding globulin's binding site. (C) Molecular docking interaction of glyphosate with rats' transthyretin. The molecular docking score is −4.29 kcal/mol. Glyphosate interacted by H-donor bonds with ASP61(2) and H-acceptor bonds with SER8 and ASP61 residues in the transthyretin's binding site.

Fig. 6

(A) Molecular docking interaction of soyasaponin ii with rats' caspase-3. The molecular docking score is −7.95 kcal/mol. Soyasaponinii interacted by H-donor bonds with LYS229 and H-acceptor bonds with LEU230, GLU231, TYR276, and LYS229 residues in caspase-3's binding site. (B) Molecular docking interaction of soyasaponin i with rats' tumor necrosis factor receptor superfamily member 1A (TNFRSF1A). The molecular docking score is −8.01 kcal/mol. Soyasaponini interacted by H-donor bonds with THR139 and GLU138 and H-acceptor bonds with LYS103, GLN111, ARG106(3), and ARG401. (C) Molecular docking interaction of soyasaponin i with rats' tumor necrosis factor receptor superfamily member 1B (TNFRSF1B). The molecular docking score is −8.40 kcal/mol. Soyasaponini interacted by H-donor bonds with MET458 and ASP455 and H-acceptor bond with GLY457 residues in TNFRSF1B's binding site.