Probiotics as an Adjunct Ameliorates Ovarian Toxicity in Endotoxemic Mice via Modulating TLR 4/MyD88/NF-κB Signalling Pathway: Insights from In Vivo and In Silico Study.

Gut microbiota plays a critical role in maintaining reproductive homeostasis, and mounting evidence highlights probiotic supplementation as a promising therapeutic candidate owing to its immunomodulatory potentials. Neurotensin (NTS), a tridecapeptide neuropeptide, has been shown to link with the regulation of inflammation and reproductive processes. This study aims to evaluate the possible simultaneous ameliorative effect of NTS receptor 1 agonist PD149163 co-administration with multi-strain probiotics in lipopolysaccharide/LPS induced ovarian dysfunction. Female Swiss albino mice (8 weeks old) were randomly assigned to seven groups: control, LPS (1 mg/kg bw), LPS + PD149163 (50 µg/kg bw), LPS + probiotics (0.6 gm/kg bw/day), LPS + PD149163 + probiotics, PD149163, and probiotics. After 32 days, plasma and ovarian samples were collected for biochemical and histological analyses. Additionally, an in-silico approach was employed to assess the potential interaction of probiotic-derived metabolites (butyrate and propionic acid) with the key proteins of TLR4/MyD88/NF-κB signalling pathway (TLR4/MD-2 complex, MyD88 and NF-κB). Co-administration of PD149163 with multi-strain probiotics attenuated inflammatory markers (NF-κB, TNF-α, IL-6), restored anti-oxidant enzyme activity (SOD, CAT), reduced lipid peroxidation (LPx), normalized hormonal levels (NTS, LH, FSH, E2) and improved ovarian histopathological features. Co-supplementation of probiotics with PD149163 as an adjunct therapy has shown superior efficacy in mitigating the ovarian dysfunction compared to employing single treatment approach. This ameliorative effect is presumably mediated by suppressing TLR4/MyD88/NF-κB signalling pathway, thereby dampening inflammatory cascade and alleviating ovarian dysfunction. Therefore, further investigations are warranted to unravel the underlying mechanisms of probiotic action on reproductive physiology, thereby providing therapeutic insights for the management of sepsis-related reproductive dysfunction.