Inhibition of neuroinflammation and pyroptosis by Fo-Shou-San enhances cognitive function in rats with vascular dementia
Description
Context：Fo-Shou-San (FSS) is a well-known traditional Chinese medicine formula used for the treatment of gynecological and cerebrovascular diseases. However, the specific molecular mechanisms underlying its therapeutic effects on vascular dementia (VaD) remain unclear. Objective: Our objective is to elucidate the mechanistic role of FSS in anti-vascular dementia by suppressing both in vivo and in vitro inflammatory responses and pyroptosis. Material and methods: We utilized the bilateral vertebral artery occlusion (2-VO) method to establish a VaD rat model and evaluated the therapeutic effects of FSS. Cognitive improvement in VaD rats following FSS treatment was assessed through behavioral experiments. Neuronal damage in the hippocampal region was observed using Nissl staining and hematoxylin-eosin (HE) staining. The expression of inflammatory cytokine genes and microglial cell activation in the rat hippocampal region was evaluated using RT-qPCR and quantitative immunohistochemistry. TUNEL staining, RT-qPCR, and Western blot techniques were employed to assess pyroptosis in the rat hippocampal tissue. Subsequently, based on high-performance liquid chromatography (HPLC) analysis, FSS derivatives were screened through molecular docking to identify the optimal compound that binds to the key pyroptosis target Caspase-1. Finally, in vitro validation demonstrated that FSS derivatives could inhibit the inflammatory response and pyroptosis induced by lipopolysaccharide (LPS) + adenosine triphosphate (ATP), in accordance with the results of molecular docking analysis. Results: FSS improved cognitive impairment and neuronal damage in VaD rats established by 2-VO, while also inhibiting the inflammatory response and pyroptosis in the rat hippocampal tissue. Building upon the results of HPLC analysis, molecular docking predicted that FSS derivatives, such as Levistilide A (LA), Imperatorin (IMP), Caffeic Acid (CA), and Ferulic Acid (FA), demonstrated optimal binding affinity with the crucial pyroptosis target, Caspase-1. Validation through in vitro experiments revealed that LA and IMP significantly inhibited the inflammatory response and pyroptosis in LPS+ATP-induced BV2 cells. Discussion and conclusion: Our study indicates that FSS can exert neuroprotective effects on VaD rats by inhibiting inflammatory responses and pyroptosis. The active components in FSS, namely LA, and IMP, demonstrated significant inhibition of the inflammatory response and pyroptosis in LPS+ATP-induced BV2 cells. This suggests the potential therapeutic value of FSS in VaD treatment and provides a novel approach for the development of Caspase-1 inhibitors in clinical settings.
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We utilized the bilateral common carotid artery ligation (BCCAL) method to establish a VaD rat model and evaluated the therapeutic effects of FSS. Cognitive improvement in VaD rats following FSS treatment was assessed through behavioral experiments. Neuronal damage in the hippocampal region was observed using Nissl staining and hematoxylin-eosin (HE) staining. The expression of inflammatory cytokine genes and microglial cell activation in the rat hippocampal region was evaluated using RT-qPCR and quantitative immunohistochemistry. TUNEL staining, RT-qPCR, and Western blot techniques were employed to assess pyroptosis in the rat hippocampal tissue. Subsequently, based on high-performance liquid chromatography (HPLC) analysis, FSS derivatives were screened through molecular docking to identify the optimal compound that binds to the key pyroptosis target Caspase-1. Finally, in vitro validation demonstrated that FSS derivatives could inhibit the inflammatory response and pyroptosis induced by lipopolysaccharide (LPS) + adenosine triphosphate (ATP), in accordance with the results of molecular docking analysis.