Exploring the mechanism of Bushen Kaiqiao Formula in alleviating neuroinflammation in ADHD rats via the NF-κB-NLRP3 pathway
ZHANG Jia-qi
SUN Ru-xin
YANG Yu-ting
ZHU Kang-lin
WANG Jing
NI Xin-qiang
HUANG Min
Abstract：Objective To investigate the therapeutic effects of Bushen Kaiqiao Formula(BSKQF)on attention deficit and spatial memory impairment in spontaneously hypertensive rats(SHRs),a widely used animal model for atten-tion-deficit hyperactivity disorder(ADHD).The potential neuroprotective mechanism was explored with a focus on mi-croglial activation under oxygen-glucose deprivation/reoxygenation(OGD/R)conditions and regulation of neuroinflam-mation via the NF-κB/NLRP3 inflammasome signaling pathway.Methods Fifty SHRs were randomly divided into a model group(SHR),low-dose(LOW),medium-dose(MID),high-dose(HIGH)BSKQF treatment groups,and a methylphenidate(MPH)group(n=10 each).Wistar-Kyoto(WKY)rats served as the control group.After 4 weeks of gavage,behavioral changes were assessed using the Y-maze test.Immunofluorescence was used to assess molecular indi-cators in the prefrontal cortex.Primary microglia were divided into control,OGD/R model,BSKQF serum-treated(LOW,MID,HIGH),and MPH groups.CCK-8,Western blot,TMRE mitochondrial potential probes,and immuno-fluorescence assays were used to detect key molecular and cellular markers in microglia and neurons.Results Compared to SHRs,BSKQF treatment significantly increased the spontaneous alternation rate in Y-mate tests(0.484 0 vs.0.628 0、0.484 0 vs.0.660 0、0.484 0 vs.0.662 0,P＜0.001).In the prefrontal cortex of SHRs,iNOS,IL-1β,and IL-6 levels were significantly elevated,while BSKQF treatment significantly reduced these markers[i-NOS:1.801 vs.1.358、1.303、1.175,P＜0.001;IL-1β(pg/mL):44.72±3.190 vs.31.65±1.348、20.85±0.934、16.21±0.723,P＜0.001;IL-6(pg/mL):13.180±0.655 vs.7.527±0.095、5.457±0.170、3.757±0.076,P＜0.001].After treat-ment with BSKQF and MPH,the expression levels of p-NF-κB-p65,p-IκBα and NLRP3 in microglia were reduced accompanied by an increase in mitochondrial membrane potential(NLRP3:1.360±0.148 vs.0.798±0.068、0.654±0.052、1.097±0.077,P＜0.05;p-NF-κB-p65:1.301±0.087 vs.0.557±0.064、0.775±0.051、0.943±0.093,P＜0.001;p-IκBα:1.097±0.117 vs.0.722±0.051、0.725±0.049、0.760±0.031,P＜0.001);mito-chondrial membrane potential(36.48±2.50 vs.74.26±2.14、79.46±2.01、101.5±0.91,P＜0.001).Medium from BSKQF-treated groups showed significantly reduced apoptosis and preserved synaptic integrity(56.75±1.15 vs.47.52±0.08、45.73±1.97、43.63±0.45,P＜0.001).Conclusion BSKQF exerts anti-inflammatory and neuro-protective effects in ADHD-like rats by modulating microglial activation and inhibiting the NF-κB/NLRP3 inflamma-some pathway,thereby improving behavioral deficits associated with ADHD.
Keywords：attention deficit hyperactivity disorderNF-κB/NLRP3 inflammasome signaling pathwayprimary microgliaoxygen-glucose deprivation/reoxygenationBushen Kaiqiao Formula
Publication Date：2025-06-15
Online Publishing Date：2025-08-18(First online date of this platform, not the publication date of the document)
Pages：9( 801-809 )