Fig 2: Schematic illustration showing the influence of BPA on the cross-talk between neuroinflammation and autophagy mediated initiation of pyroptosis and progression of anxiety-related behaviors in postnatal male rats. BPA exposure resulted in a remarkable elevation in the level of proinflammatory cytokines particularly IL-2 and IL-12 that are significant for differentiation and function of T-cells. The Canonical NLRP3 inflammasome activation requires two steps: the first is priming step and the second is activation step. In the priming step, toll-like receptors (TLR) stimulation promotes the transcription and expression of NLRP3 and pro-IL-1β through NF-κB. Subsequently, various DAMPs and PAMPs induce the activation step by initiating numerous cellular and molecular cascades, including K+ efflux, Ca2+ influx, mitochondrial dysfunction, reactive oxygen species (ROS) release, and lysosomal disruption. The NLRP3-dependent self-cleavage and activation of pro-caspase-1 self-cleavage and activation leads to the maturation of the pro-inflammatory cytokines IL-1β and IL-18. Additionally, gasdermin D (GSDMD) is cleaved by activated caspase-1, releasing its N-terminal domain, which then integrates into the cell membrane to create pores. These pores allow the release of cellular contents, including IL-1β and IL-18, and trigger pyroptosis. BPA exposure caused a significant elevation in the expression level of NF-κB, IL-1β, NLRP3 and caspase-1. The release of DAMPs and PAMPs is a combined by an increase in extracellular glutamate levels, which further leads to excitotoxic neuron damage through NMDA receptors. Furthermore, glutamate-mediated Ca2+ influx occurs through NMDA receptors. mTOR is a downstream target of PI3K and AKT pathway which could be downregulated by apoptotic signals, therefore, induction of autophagy by inhibition of the mTOR may result in dysregulated autophagic machinery led to neuronal degeneration. Damaged and dying neurons secrete DAMPs besides prevalence of cytokines all together lead to activation of microglia via binding to cell surface receptors (e.g. TLR), resulting in activation of the NF-κB pathway. This activation induces a signaling cascade, causing enhancement in the expression of cytokines such as pro-IL-1β and pro-IL-18 as well as NLRP3. Autophagy begins with the formation of double-membrane vesicles known as autophagosomes. Autophagosomes combine with lysosomes to generate autophagolysosomes, which degrade their contents into basic molecules that can be recycled for reuse. The elevated beclin-1, LC3A, and LC3B levels by inhibiting the PI3K/Akt/mTOR signaling pathway, triggering autophagy in brain tissue. Once microglia get activated, they become polarized to an M1, pro-inflammatory phenotype and secrete pro-inflammatory cytokines and factors as well as, activating astrocytes. Activation and dysfunction of astrocytes contributes to BBB, allowing infiltration of different immunological cells. Note At the end of each line, the arrowhead indicates activating the neural projection, while short line for inhibition

Fig 3: The molecular docking analysis of BPA and neuroinflammatory, autophagic as well as, pyroptotic molecules displaying 2D and 3D binding interactions of BPA [PubChem CID: 6623] against A NF-kB [PDB ID: 1NFK], B IL-1β [PDB ID: 2MIB], C IL-2 [PDB ID: 4YQX], D IL-12 [PDB ID: 3HMX], E COX-2 [PDB ID: 1PXX], F NLRP3 [PDB ID: 7vtq], G Beclin-1 [PDB ID: 2PON], H LC3A [PDB ID: 6TBE], I LC3B [PDB ID: 5XAC], J Caspase-1 [PDB ID: 6VIE]. The green dotted lines denote hydrogen bonds between ligand and aminoacids, whereas bink/purple dotted lines represent hydrophobic interactions. Electrostatic interactions are shown as orange dotted lines. The red dotted line indicate an unfavorable donor-donor

Fig 4: Effect of postnatal exposure to BPA (50 and 125 mg/kg/day) on the expression level of NF-kB (A and B), IL-1β (C and D), IL-2 (E and F), IL-12 (G and H), COX-2 (I and J) in the PFC and hippocampusof PND60 and PND95 male rats. Inflammatory markers for each dose and age plotted on axes 1 and 2 of a Principal Coordinates (PCO) graph by computing the distance between centroids based on the groups in the PFC and hippocampus (K and L). The results are expressed as the mean ± SE (n = 6 rats/group). Groups with different superscript letters means there is a significant change between them at P < 0.05. Statistical significance test for comparison was done by MANOVA, followed by post hoc Tukey’s HSD multiple comparison test

Fig 5: Safranal inhibited the inflammatory response by regulating the Sirt1/NF-κB pathway. A Western blotting was conducted to assess the protein levels of factors involved in the Sirt1/NF-κB pathway. Quantification of Sirt1 (B), p-p65/p65 (C), and p-IκB/IκB (D) levels; E immunofluorescence staining of Sirt1 was performed to measure Sirt1 expression in the kidney tissues of each group (scale bar = 20 µm). F–H ELISA was performed to measure the levels of inflammatory factors (IL-2, IL-6, and TNF-α) in renal homogenates. ***P < 0.001 vs. the sham group; &P < 0.05, &&P < 0.01, &&&P < 0.001 vs. the MGN group. n.s. not significant. #P < 0.05 vs. the MGN + Saf-100 group. n = 3