Deciphering the Role of NLRP-3/Caspase-1/GSDMD Pyroptotic Signal, miR-675-5p, and miR-1247-5p in Mitigation of Neurobehavioral and Neuropathological Alterations in Rotenone-Induced Striatal Neurodegeneration by Vitex agnus-castus Leaf Extract and/or

Rotenone (ROT ) exposure causes behavioral and motor abnormalities, including bradykinesia, catalepsy, and unsteady gait, as in Parkinsonism. Vitex agnus-castus (Vitex A-C) has been extensively utilized in the management of various female ailments besides its role as an agonist for D2 dopaminergic receptors. Pramipexole (Prami) is a dopamine agonist (DA) receptor , which can reduce complications of dopamine therapy. Therefore, this investigation aimed to assess the possible ameliorating effects of Vitex A-C and/or Prami against ROT-evoked striatal neurodegeneration, as well as to shed light on the possible underlying mechanisms . Seventy adult male albino rats were allocated into seven groups (n = 10 rats/group): Group I (control group), Group II (Prami control group), Group III (Vitex A-C control group), Group IV (ROT group), while Groups V-VII were injected with an intraperitoneal injection of ROT along with a daily oral administration of Vitex A-C orPrami, or their combination, respectively, for 60 days. Molecular docking results showed that Vitexin complements with superior performance in α-synuclein (- 5.3 vs - 3.4 kcal/mol), caspase-1 (- 6.9 vs - 4.6 kcal/mol), and NF-κB p65 (- 6.8 vs - 4.5 kcal/mol) targeting. Agnuside's dopaminergic and anti-inflammatory effects, with Vitexin's anti-aggregation and anti-inflammatory properties. Our results indicated that Vitex A-C and/or Prami markedly ameliorated ROT-induced striatal neurodegeneration, evidenced by their abilities to mitigate ROT-triggered neurobehavioral alterations, dopamine, oxidative stress (MDA), antioxidant (GPX and catalase), and inflammatory markers (NF-kB P65, IL-1β). Vitex A-C and/or Prami-treated groups decreased pyroptotic signal as evidenced by a remarkable decline in the protein expression of NLRP3, caspase-1, and GSDMD; gene expression of ASC; and tissue levels of IL-1β and IL-18. Additionally, Vitex A-C and/or Prami substantially downregulated α-synuclein and upregulated TH protein expressions. On the molecular levels, the combination group rectified ROT-triggered dysregulations in the expressions of HMGB1, AIF-1, and miR-1247-5p without any significant impact on miR-675-5p. The combined therapy showed an improvement in striatal histoarchitecture with mitigation of caspase-1 and glial fibrillary acidic protein immunoreactivity with an upregulation in synaptophysin immunoreactivity. In conclusion, the combined therapy of Vitex A-C and Prami holds a promising therapeutic avenue over them alone against ROT-associated striatal neurodegeneration via inhibiting the pyroptotic pathway.