ABSTRACT
Objective To explore the effect of scutellarin on lipopolysaccharide(LPS)induced neuroinflammation in BV-2 microglia cells.Methods BV-2 microglia were cultured and randomly divided into 6 groups:control group(Ctrl),cyclic GMP-AMP synthetase(cGAS)inhibitor RU320521 group(RU.521 group),LPS group,LPS+RU.521 group,LPS+scutellarin pretreatment group(LPS+S)and LPS+S+RU.521 group.The expressions of cGAS,stimulator of interferon gene(STING),nuclear factor kappa B(NF-κB),phosphorylated NF-κB(p-NF-κB),neuroinflammatory factors PYD domains-containing protein 3(NLRP3)and tumor necrosis factor α(TNF-α)in BV-2 microglia were detected by Western blotting and immunofluorescent double staining(n= 3).Results Western blotting and immunofluorescent double staining showed that compared with the control group,the expression of cGAS,STING,p-NF-κB,NLRP3 and TNF-α in BV-2 microglia increased significantly after LPS induction(P<0.05),while the expression of cGAS,STING,p-NF-κB,NLRP3 and TNF-α in LPS+S group were significantly lower than those in LPS group(P<0.05).Treatment with cGAS pathway inhibitor RU.521 showed similar effects as the pre-treatment group with scutellarin.In addition,the change of NF-κB in each group was not statistically significant(P>0.05).Conclusion Scutellarin inhibits the neuroinflammation mediated by BV-2 microglia cells,which may be related to cGAS-STING signaling pathway.
ABSTRACT
Objective To investigate the effect oftopiramate on NACHT-LRR-PYD-containing protein 3 (NALP3) inflammasome and intedeukin (IL)-1β levels in trigeminal ganglion of migraine rats.Methods Forty adult male Sprague Dawley rats were randomly divided into blank group,saline group,model group,prevention control group,10 mg/kg topiramate group,30 mg/kg topiramate group,60 mg/kg topiramate group,and 90 mg/kg topiramate group (n=5).Inflammatory soup was used to stimulate the dual matter of rats repeatedly to induce migraine models:rats in the blank group were without any treatment,those in the saline group were given saline to stimulate the dual matter,different concentrations of topiramate group were given to migraine models of the 10 mg/kg topiramate group,30 mg/kg topiramate group,60 mg/kg topiramate group,and 90 mg/kg topiramate group,respectively,and migraine models of the prevention control group were given saline containing 1％ Tween80.Three h after the last treatment,the expressions of NALP3,caspase-1 precursor (pro-caspase-1),caspase-1 and IL-1β in trigeminal ganglion of rats were detected by Western blotting,and the best concentration oftopiramate was chosen for subsequent immunofluorescence experiments.Ten healthy male adult SD rats were randomly divided into control group and topiramate group (n=5);the expression levels of NALP3,caspase-1 and IL-1β in trigeminal ganglion of the two groups were detected by immunofluorescence.Results The expression levels ofNALP3,pro-caspase-1,caspase-1 and IL-1β were not significantly different between saline group and blank group (P＞0.05);the expression levels ofNALP3,pro-caspase-1,caspase-1 and IL-1β in the model group were significantly higher than those in the saline group (P＜0.05).The expression levels of NALP3,pro-caspase-1,caspase-1 and IL-1β in 60 mg/kg topiramate group and 90 mg/kg topiramate group were significantly lower than those in the prevention control group (P＜0.05).The fluorescence intensity ofNALP3,caspase-1 and IL-1β in topiramate group was significantly lower than that in control group.Condusion Topiramate can inhibit the expression of NALP3 inflammasome and IL-1β in the trigeminal ganglion of migraine rat models,and it is likely to be one of the important mechanisms for the prevention of migraine.