# Astragaloside IV ameliorates atrazine-induced male reproductive toxicity: an in vivo and in silico analysis

## TL;DR

Astragaloside IV helps protect against atrazine's harmful effects on male mouse reproduction by reducing oxidative stress and inflammation.

## Contribution

This study is the first to demonstrate astragaloside IV's protective effects against atrazine-induced male reproductive toxicity using in vivo and in silico methods.

## Key findings

- Astragaloside IV improved antioxidant markers and testosterone levels in atrazine-exposed mice.
- Molecular docking showed astragaloside IV strongly interacts with proteins involved in oxidative stress and inflammation.
- Histopathological improvements were observed in testes of mice treated with astragaloside IV.

## Abstract

Atrazine (ATZ) stands as the most widely utilized herbicide globally and is known for its adverse impacts on the reproductive system. Although astragaloside IV (AS IV) is well known for possessing various health benefits, its protective effects against ATZ-induced toxicity remain unexplored. This study aimed to investigate the ameliorative potential of AS IV against ATZ-induced male reproductive toxicity in mice.

Eight-week-old CD-1 mice were allocated into four groups (n = 10). ATZ and AS IV were administered at doses of 100 mg/kg/day and 40 mg/kg/day, respectively. Treatments were continued for 21 days, after which the animals were sacrificed for plasma biochemical analyses and testes collection for histopathological examination. One-way analysis of variance (ANOVA) followed by Bonferroni’s multiple comparison test was used for data analysis. Molecular docking studies were performed to evaluate ATZ and AS IV interactions with oxidative stress- and inflammation-related proteins, including glutathione (GSH), glutathione peroxidase (GPx), superoxide dismutase (SOD), and Nrf2, NF-κβ, IL-1β, IL-6, TNF-α, cullin-3, and Keap-1.

Biochemical analysis revealed significant reductions in GSH levels (p < 0.001), SOD activity (p < 0.001), and GPx activity (p < 0.05), along with elevated malonaldehyde levels (p < 0.01), following ATZ exposure. AS IV treatment in ATZ-exposed mice significantly improved these markers (p < 0.05). ATZ exposure led to significant decreases in testosterone (p < 0.001) and androgen-binding protein (ABP) levels (p < 0.001) within the ATZ group, whereas AS IV supplementation significantly improved these markers (p < 0.05). Histopathological examination revealed sloughed and collapsed seminiferous epithelia with vacuoles and poorly formed spermatids in ATZ-exposed mice, which were mitigated by AS IV treatment. The docking study revealed ATZ’s moderate interactions with key oxidative stress and inflammation-related proteins (binding energies: −4.7 to −5.5 kcal/mol), with glutathione (GSH) (−5.5 kcal/mol) showing the strongest binding. Notable stabilizations include SOD (three hydrogen bonds) and modulation of antioxidant (SOD, Nrf2) and anti-inflammatory (IL-1β and TNF-α) pathways. Moreover, AS IV demonstrated significant binding interactions with GSH (−9.2 kcal/mol), cullin-3 (−9.1 kcal/mol), and keap-1 (−8.9 kcal/mol). Molecular dynamics (MD) simulations showed strong stability for GPx and IL-1β targets against ATZ, and AS IV exhibited strong stability for GSH and cullin-3.

AS IV appears to be a promising natural compound for preventing ATZ-induced male reproductive toxicity. Further investigations to elucidate the molecular mechanisms behind such positive effects are warranted.