Integrated In-silico and In-vivo Assessments of Betaine's Effect on the Hypothalamic-Pituitary-Testicular (HPT) Axis in Fluoride-Treated Rats

Toxicity is associated with undue sodium fluoride (NaF) exposure, and Betaine (BET) is recognised for its nutraceutical benefits. Although it is necessary to reduce toxic level exposure to fluoride, the literature lacks information on the role of BET in mitigating fluoride-induced reproductive toxicity. Therefore, this study assesses the impact of BET on NaF-induced reproductive perturbation in male rats. Wistar rats were treated with NaF (9 mg/kg) alone or co-treated with BET (50 or 100 mg/kg) for 28 d. Our findings indicate that BET significantly mitigated alterations in sperm functionality indices caused by NaF treatment. BET substantially increased reproductive hormone levels and averted NaF-induced increases in oxidative stress biomarkers and testicular enzymes. NaF-induced increases in inflammatory markers in the testis, epididymis, and hypothalamus were effectively reversed upon BET co-treatment. Also, co-treatment with BET protected genome integrity, as evidenced by p53 and apoptotic markers Bax and Bcl-2 levels, abating damages in the testes, epididymis, and hypothalamus of NaF-treated rats. Also, our findings from in-silico studies revealed that BET moderately inhibits the molecular activation of the inhibitor of nuclear factor-kappa B kinase, hypoxia-inducible factor -1 alpha, and proviral integration for the Moloney murine leukaemia virus-1 kinase. While it is preferable to reduce fluoride exposure, the relevant findings here indicate that BET exhibits anti-inflammatory, antioxidant, and anti-apoptotic properties that ameliorate inadvertent NaF-mediated toxicities in experimental rats exposed to NaF.