Impact of Liparis nervosa extract on neuroinflammation mediated by LPS-induced BV-2 microglial cells and its bioactive components analysis
To investigate the impact and potential mechanisms of extracts from different parts of Liparis nervosa on neuroinflamma-tion by lipopolysaccharide(LPS)-induced BV-2 microglial cells.The materials of L.nervosa were subjected to crushing,ethanol ex-traction,and concentration to obtain an alcohol extract.Subsequently,the extract was further extracted to obtain petroleum ether ex-tract,ethyl acetate extract,N-butanol extract,and aqueous phase extract.The ethyl acetate extract was separated into distillate ①-⑥using D101 macroporous resin column chromatography.The experiment was divided into control group,LPS model group,L.nervosa extract group,and LPS+L.nervosa group.LPS was utilized to induce a neuroinflammatory cell model in BV-2 microglial cells.The Griess test was utilized for detecting the production of nitric oxide(NO)in the cell supernatant.Cell viability was detected by MTT as-say.The release of interleukin-6(IL-6)and tumor necrosis factor alpha(TNF-α)in the cell supernatant was quantified using ELISA.RT-qPCR was utilized to assess the mRNA levels of pro-inflammatory cytokines inducible nitric oxide synthase(iNOS),cyclooxyge-nase-2(COX-2),interleukin(IL)-6,IL-1β,and TNF-α.The protein expression of iNOS,COX-2,nuclear factor kappa-B p65(p65),p-p65,extracellular signal-regulated kinase(ERK),p-ERK,c-jun N-terminal kinase(JNK),p-JNK,p38 mitogen-activated protein kinase(p38),and p-p38 MAPK(p-p38)were also evaluated by Western blot.The chemical composition of active substances in L.nervosa was analyzed using the UHPLC-Q-Exactive Orbitrap technology and literature comparison.Our findings indicate that ex-tracts from different parts of L.nervosa exhibit a significant reduction in the release of NO from LPS-induced BV-2 microglial cells.Specifically,the ethyl acetate extract demonstrates the most notable inhibitory effect without causing cell toxicity.Additionally,the dis-tillate ⑥ extracted from the ethyl acetate exhibits a reduction in the mRNA and protein levels of iNOS,COX-2,IL-6,IL-1β,and TNF-α in a dose-dependent manner,and it inhibits the protein expression of p-p65,p-ERK,p-p38,and p-JNK in LPS-induced BV-2 microglial cells.A total of 79 compounds in the distillate ⑥ were identified by mass spectrometry,including 12 confirmed compounds with anti-inflammatory effects.This study confirmed the remarkable efficacy of L.nervosa extract in the treatment of neuroinflamma-tion,which may be achieved through the inhibition of NF-κB and MAPK signaling pathways.
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